Synergistic Renoprotective Effects of Green Tea Extract and Gemigliptin against Tacrolimus-Induced Nephrotoxicity in Mice / Efectos Renoprotectores Sinérgicos del Extracto de Té Verde y Gemigliptina contra la Nefrotoxicidad Inducida por Tacrolimus en Ratones

SUMMARY: Although tacrolimus (TAC) significantly reduces allograft rejection incidence in solid-organ transplantation, its long-term use is associated with an increased risk of TAC-induced nephrotoxicity. In this study, we investigated the renoprotective effects of green tea extract (GTE) with or without the dipeptidyl peptidase 4 inhibitor, gemigliptin, by assessing serum creatinine levels, the amount of proteinuria, and histopathology in TAC-induced nephrotoxicity. TAC-induced nephrotoxicity was induced by intraperitoneal TAC injection, GTE was administered via subcutaneous injection, and gemigliptin was administered orally. Mice with TAC-induced nephrotoxicity exhibited a significant increase in both serum creatinine levels and 24-hour urine protein. However, when treated with GTE via subcutaneous injection, mice showed a decrease in serum creatinine levels and the amount of proteinuria. When GTE was combined with gemigliptin, further renoprotective effects were observed in biochemical assessments, consistent with the attenuation of TAC-induced nephrotoxicity in histopathology. The expression of p53 protein was lower in the mice treated with the combination of GTE and gemigliptin compared to mice with TAC-induced nephrotoxicity. Our results demonstrate that the combination of GTE and gemigliptin treatment reveals synergistic renoprotective effects by decreasing the expression of p53 protein. These findings suggest that the combination of GTE and gemigliptin could potentially be used as a prophylactic or therapeutic strategy for TAC-induced nephrotoxicity.

Aunque tacrolimus (TAC) reduce significativamente la incidencia de rechazo de aloinjertos en trasplantes de órganos sólidos, su uso a largo plazo se asocia con un mayor riesgo de nefrotoxicidad inducida por TAC. En este estudio, investigamos los efectos renoprotectores del extracto de té verde (GTE) con o sin el inhibidor de la dipeptidil peptidasa 4, gemigliptina, mediante la evaluación de los niveles de creatinina sérica, la cantidad de proteinuria y la histopatología en la nefrotoxicidad inducida por TAC. La nefrotoxicidad inducida por TAC se indujo mediante inyección intraperitoneal de TAC, el GTE se administró mediante inyección subcutánea y la gemigliptina se administró por vía oral. Los ratones con nefrotoxicidad inducida por TAC mostraron un aumento significativo tanto en los niveles de creatinina sérica como en la proteína en orina de 24 horas. Sin embargo, cuando se trataron con GTE mediante inyección subcutánea, los ratones mostraron una disminución en los niveles de creatinina sérica y en la cantidad de proteinuria. Cuando se combinó GTE con gemigliptina, se observaron efectos renoprotectores adicionales en las evaluaciones bioquímicas, lo que concuerda con la atenuación de la nefrotoxicidad inducida por TAC en histopatología. La expresión de la proteína p53 fue menor en los ratones tratados con la combinación de GTE y gemigliptina en comparación con los ratones con nefrotoxicidad inducida por TAC. Nuestros resultados demuestran que la combinación de tratamiento con GTE y gemigliptina revela efectos renoprotectores sinérgicos al disminuir la expresión de la proteína p53. Estos hallazgos sugieren que la combinación de GTE y gemigliptina podría usarse potencialmente como estrategia profiláctica o terapéutica para la nefrotoxicidad inducida por TAC.

Perfil molecular de cáncer de endometrio en mujeres de Bogotá DC, Colombia / Molecular profile of endometrial cancer in women from Bogota DC, Colombia

Introducción: El cáncer de endometrio ocupa el sexto lugar en incidencia del cáncer en mujeres. La caracterización molecular de este cáncer permite optimizar la estratificación de riesgo para mejorar el tratamiento de las pacientes. Objetivo: Determinar el perfil molecular TCGA de pacientes con cáncer de endometrio en Bogotá, D.C., Colombia. Método: Estudio descriptivo en una cohorte de pacientes con cáncer de endometrio. Las mutaciones en los exones 9 a 14 del gen POLE fueron identificadas mediante amplificación por reacción en cadena de la polimerasa, seguida de secuenciación Sanger y análisis bioinformático. La expresión de las proteínas MMR y p53 se identificó mediante inmunohistoquímica. Resultados: Se incluyeron 40 pacientes con una mediana de edad de 66 años. El 15% presentaron mutaciones en el dominio exonucleasa de POLE. El 32% de las pacientes que no presentaron mutaciones manifestaron deficiencia en el sistema MMR. El 43,47% de las pacientes sin mutaciones en POLE ni alteración del sistema MMR presentaron alteración de la proteína p53. Conclusiones: La población de cáncer de endometrio analizada presenta un perfil molecular TCGA similar a lo reportado para otras poblaciones.

Introduction: Endometrial cancer ranks sixth in cancer incidence among women. Its molecular characterization allows for a more precise risk stratification with the aim of improving patient treatment. Objective: To determine the TCGA molecular profile of patients with endometrial cancer in Bogota, Colombia. Method: A descriptive study of a cohort of patients with endometrial cancer. The expression of MMR proteins and p53 was identified through immunohistochemistry. Mutations in exons 9 to 14 of the POLE gene were identified through polymerase chain reaction amplification, followed by Sanger sequencing and bioinformatic analysis. Results: Forty patients were included in the study, with a median age of 66 years, 15% of them exhibited mutations in the exonuclease domain of POLE, while 32% of patients without mutations showed deficiency in the MMR system. Forty three percent of patients without mutations in POLE or MMR alterations showed aberrant p53 protein expression. Conclusions: The analyzed population of endometrial cancer presents a TCGA molecular profile similar to that reported for other populations.

α-Hederin Induces Apoptosis in Hepato-cellular Carcinoma Cells by Activating and Stabilizing p53/Noxa Signaling Pathway / 医药导报

Objective To investigate the inhibitory effects and mechanisms of α-hederin,an active ingredient in Fruc-tus Akebiae,on hepatocellular carcinoma(HCC)cells.Methods HCC cells were divided into four groups and treated with α-hederin(0,10,20,and 30 μmol·L-1)for 24 h and 48 h,respectively.MTT assays were used to detect the cell proliferation rate,flow cytometry(FCM)was used to detect the apoptotic rate,transcriptomics was used to screen signaling pathways in α-hederin-treated HCC cells,RNA interference was exploited to verify the underlying signaling pathway,and real-time quantitative PCR(qRT-PCR)and Western blotting(WB)were used to detect expression changes of the mRNA and protein of TP53(p53),PMAIP1(Noxa),and apoptosis-associated proteins,Caspase9 and Caspase3.Results α-Hederin induced apoptosis by activa-ting apoptosis-associated proteins,PARP,Caspase9 and Caspase3.Transcriptomics,qRT-PCR,and WB results also showed that α-hederin increased the mRNA and protein expression of p53 and Noxa.Furthermore,α-hederin inhibited the protein degradation of p53 and Noxa,reversing the apoptosis decrease in p53/Noxa siRNA-knocked-down HCC cells.In vivo results showed that α-hederin inhibited the growth of HCC tumors.Conclusion α-hederin may induce the apoptosis of HCC cells by activating and stabilizing the p53/Noxa signaling pathway.

Mechanism of esculetin promoting ferroptosis of mouse breast cancer 4T1 cells based on p53/SLC7A11/GPX4 axis / 中国病理生理杂志

AIM:To investigate the mechanism through which esculetin induces ferroptosis of mouse breast cancer 4T1 cells.METHODS:Molecular docking of esculetin with p53,solute carrier family 7 member 11(SLC7A11)and glutathione peroxidase 4(GPX4)proteins was performed,and Kaplan-Meier curves and time-dependent receiver oper-ating characteristic curves were drawn.The 4T1 cells were divided into 6 groups,namely the control(CON),1/2 IC50,IC50,2 IC50,erastin,and capecitabine groups.The appropriate drug concentration for treating 4T1 cells was screened by CCK-8 assay.The invasion and migration abilities of 4T1 cells following esculetin treatment at the selected drug concentra-tion were analyzed by scratch test and Transwell assay.Mitochondrial morphological change were examined by electron mi-croscopy.The levels of ferroptotic cell death were then quantified by Fe2+,reactive oxygen species(ROS),malondialde-hyde(MDA),glutathione(GSH),and GPX4 assays.Western blot was performed to evaluate the protein levels of p53,SLC7A11,GPX4 and acyl-CoA synthetase long-chain family member 4(ACSL4).RESULTS:Compared with CON group,the esculetin 1/2 IC50,IC50 and 2 IC50 groups showed smaller size,increased membrane density,and decreased ridge of mitochodria,as well as decreased levels of GSH and GPX4,reduced cell invasion and migration abilities,and de-creased levels of SLC7A11 and GPX4 proteins(P＜0.05).Furthermore,these groups exhibited increased levels of Fe2+,ROS,and MDA,as well as elevated p53 and ACSL4 protein abundance(P＜0.05).CONCLUSION:Esculetin pro-motes ferroptosis of 4T1 cells through a mechanism involving the p53/SLC7A11/GPX4 regulatory axis.

Investigation on the signaling pathways in the mechanism of hyperoxia-induced acute lung injury based on transcriptomics sequencing / 中华危重病急救医学

Objective:To observe and verify the changes of transcriptome in hyperoxia-induced acute lung injury (HALI), and to further clarify the changes of pathways in HALI.Methods:Twelve healthy male C57BL/6J mice were randomly divided into normoxia group and HALI group according to the random number table, with 6 mice in each group. The mice in the normoxia group were fed normally in the room, and the mice in the HALI group was exposed to 95% oxygen to reproduce the HALI animal model. After 72 hours of hyperoxia exposure, the lung tissues were taken for transcriptome sequencing, and then Kyoto Encyclopedia of Genes and Genomes database (KEGG) pathway enrichment analysis was performed. The pathological changes of lung tissue were observed under light microscope after hematoxylin-eosin (HE) staining. Real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were used to verify the key molecules in the signal pathways closely related to HALI identified by transcriptomics analysis.Results:Transcriptomic analysis showed that hyperoxia induced 537 differentially expressed genes in lung tissue of mice as compared with the normoxia group including 239 up-regulated genes and 298 down-regulated genes. Further KEGG pathway enrichment analysis identified 20 most significantly enriched pathway entries, and the top three pathways were ferroptosis signaling pathway, p53 signaling pathway and glutathione (GSH) metabolism signaling pathway. The related genes in the ferroptosis signaling pathway included the up-regulated gene heme oxygenase-1 (HO-1) and the down-regulated gene solute carrier family 7 member 11 (SLC7A11). The related genes in the p53 signaling pathway included the up-regulated gene tumor suppressor gene p53 and the down-regulated gene murine double minute 2 (MDM2). The related gene in the GSH metabolic signaling pathway was up-regulated gene glutaredoxin 1 (Grx1). The light microscope showed that the pulmonary alveolar structure of the normoxia group was normal. In the HALI group, the pulmonary alveolar septum widened and thickened, and the alveolar cavity shrank or disappeared. RT-RCR and Western blotting confirmed that compared with the normoxia group, the mRNA and protein expressions of HO-1 and p53 in lung tissue of the HALI group were significantly increased [HO-1 mRNA (2 -ΔΔCt): 2.16±0.17 vs. 1.00±0.00, HO-1 protein (HO-1/β-actin): 1.05±0.01 vs. 0.79±0.01, p53 mRNA (2 -ΔΔCt): 2.52±0.13 vs. 1.00±0.00, p53 protein (p53/β-actin): 1.12±0.02 vs. 0.58±0.03, all P < 0.05], and the mRNA and protein expressions of Grx1, MDM2, SLC7A11 were significantly decreased [Grx1 mRNA (2 -ΔΔCt): 0.53±0.05 vs. 1.00±0.00, Grx1 protein (Grx1/β-actin): 0.54±0.03 vs. 0.93±0.01, MDM2 mRNA (2 -ΔΔCt): 0.48±0.03 vs. 1.00±0.00, MDM2 protein (MDM2/β-actin): 0.57±0.02 vs. 1.05±0.01, SLC7A11 mRNA (2 -ΔΔCt): 0.50±0.06 vs. 1.00±0.00, SLC7A11 protein (SLC7A11/β-actin): 0.72±0.03 vs. 0.98±0.01, all P < 0.05]. Conclusions:HALI is closely related to ferroptosis, p53 and GSH metabolism signaling pathways. Targeting the key targets in ferroptosis, p53 and GSH metabolism signaling pathways may be an important strategy for the prevention and treatment of HALI.

Effects of hesperetin on memory function and apoptosis-related protein expression in senile model rats / 中华行为医学与脑科学杂志

Objective:To investigate the effect and mechanism of different doses of luteolin on memory function and apoptosis-related proteins of aging rats induced by D-galactose.Methods:Forty-eight SPF-grade male Wistar rats aged 6-8 weeks were randomly divided into control group, model group, luteolin low-dose group (25 mg/kg), medium-dose group (50 mg/kg), high-dose group (100 mg/kg), and vitamin C group (100 mg/kg), with 8 rats in each group. D-galactose (1 000 mg/kg) was subcutaneously injected to establish the aging rat model, while luteolin was used for preventive treatment. The Morris water maze test was used to evaluate the learning and memory abilities of the rats.Transmission electron microscopy was used to detect the morphology of hippocampal neurons in rats.Spectrophotometry was used to detect the levels of interleukin-6 (IL-6), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), superoxide dismutase (SOD), malondialdehyde (MDA), and the total antioxidant capacity (T-AOC). RT-PCR was used to detect miR-34a mRNA expression.Western blot technique was used to detect the expression levels of silent regulator protein 1 (SIRT1), B-cell lymphoma-2 (Bcl-2), cleaved caspase-3, p53, and p21. Statistical analysis was performed using SPSS 22.0, and one-way ANOVA was used for multi-group comparison, followed by LSD- t test for further pairwise comparisons. Results:(1) The differences in escape latency among the 6 groups of rats were statistically significant ( F=120.93, P<0.001). The latency of first finding the platform location of the model group rats ((54.61±3.60) s) was higher than that of the control group ((10.54±4.27) s) ( P<0.05). The latency of first finding the platform location of rats in the low, medium and high dosage groups of luteolin ((45.50±3.81)s, (37.46±2.94) s, (32.32±3.14) s) was lower than that of the model group ((54.61±3.60) s) (all P<0.05). (2) The differences of SOD, MDA, T-AOC, TNF-α, IL-1β, and IL-6 levels in the cerebral cortex of the 6 groups of rats were all statistically significant ( F=281.636, 75.119, 208.228, 38.999, 28.428, 52.767, all P<0.001). Compared with the control group, the model group showed abnormal levels of inflammatory factors and antioxidant indexes. In the medium and high dosage groups of luteolin, the SOD and T-AOC contents in the cerebral cortex of rats were higher than those in the model group (all P<0.05), while the levels of MDA, TNF-α, IL-1β, and IL-6 were lower than those in the model group (all P<0.05). (3) The differences in relative expression levels of miR-34a mRNA among the 6 groups of rats were statistically significant ( F=81.439, P<0.001). The expression levels of miR-34a mRNA in the hippocampal tissues of rats in the luteolin treatment group were lower than those in the model group ( P<0.05). (4) The differences in protein expression levels of SIRT1, p53, and p21 in the hippocampal tissues of the 6 groups of rats were statistically significant ( F=159.946, 38.342, 123.608, all P<0.001). The expression levels of p53 and p21 in the medium and high dosage groups of luteolin were lower than those in the model group (all P<0.05), while the expression level of SIRT1 protein was higher than that in the model group ( P<0.05). (5) The differences in protein expression levels of Bcl-2 and cleaved caspase-3 in the hippocampal tissues of the 6 groups of rats were statistically significant ( F=112.659, 43.296, both P<0.05). The expression levels of Bcl-2 in the low, medium, and high dosage groups of luteolin ((0.24±0.04), (0.40±0.03), (0.48±0.05) pg/μg) were higher than those in the model group ((0.09±0.06) μg) ( P<0.05), while the expression levels of cleaved caspase-3 in the low, medium, and high dosage groups of luteolin ((0.62±0.04), (0.61±0.09), (0.51±0.10) μg) were lower than those in the model group ((0.75±0.05) μg) ( P<0.05). Conclusions:Luteolin can alleviate cellular oxidative damage through downregulating the miR-34a SIRT1/p53 signaling pathway and reducing cell apoptosis.

Basic and clinical research progress in carbon ion therapy for colorectal cancer / 中华放射肿瘤学杂志

Colorectal cancer is a common clinical malignant tumor. As the main therapeutic method of colorectal cancer, radiotherapy has a good inhibitory effect on tumor progression. In recent years, because of its good physical and biological advantages, carbon ion has shown better clinical efficacy than traditional radiotherapy in the treatment of local recurrence or distant metastasis of colorectal cancer. In this article, basic and clinical studies related to the efficacy of carbon ion therapy for the recurrence of colorectal cancer in recent years were reviewed, aiming to provide theoretical basis for preventing and reducing adverse reactions after radiotherapy and prolonging the survival of colorectal cancer patients.

Mechanism of CD137 signal regulating P53/P21 pathway to promote senescence of vascular smooth muscle cells / 中华老年心脑血管病杂志

Objective To explore the mechanism by which CD137 signal regulates the aging of vas-cular smooth muscle cells(VSMCs).Methods Thirty 8-week-old male C57BL/6J mice were ran-domly divided into a young group(8 weeks old)and an aged group(80 weeks old),with 30 mice in each group.After corresponding periods of feeding,the mice were euthanized,and the plasma and aortic blood vessels were isolated.In the cell experiments,normal VSMCs were divided into a control group,bleomycin(BLM)group,combined agonist group,and combined inhibitor group.The cellular senescence level of VSMCs was assessed using a cellular senescence β-galactosidase staining kit.Western blotting and PCR were employed to examine the expression of senescence-related proteins in tissues and cells,while ELISA was utilized to measure the expression of senes-cence-related inflammatory factors.Results The expression of CD137 and γ-H2AX in the aorta was significantly higher,while that of PCNA was obviously lower in the aged group than the young group(P＜0.05).The plasma level of CD137 was notably higher in the aged group than the young group(154.0±4.1 pg/ml vs 98.0±2.3 pg/ml,P＜0.05).Compared with the normal control group,there were significantly more aged VSMCs in the BLM group(P＜0.05).While,treatment of combined agonist resulted in larger amount of aged VSMCs when compared with the BLM group(P＜0.05),which was reversed by combined inhibitor treatment(P＜0.05).The levels of TNF-α,IL-6 and IL-1β were significantly elevated in the BLM group than the normal control group(P＜0.05).The combined agonist group had even higher levels of TNF-α,IL-6,and IL-1βthan the BLM group(P＜0.05),but the levels were decreased in the combined inhibitor group(P＜0.05).Compared with the normal control group,the expression of Bcl-2,γ-H2AX,P53,and P21 were significantly increased in the BLM group,combined agonist group,and combined inhibi-tor group,while that of PCNA was significantly decreased(P＜0.05).Compared with the BLM group,the expression of P53 and P21 in the combined agonist group showed an increase(P＜0.05),and the expression of P53 was significantly decreased in the combined inhibitor group(P＜0.05).Conclusion CD137 signal regulates the P53/P21 pathway to promote VSMC aging.

Development of a typing detection method for high-risk human papillomavirus and related tumor suppressor genes p53 and RB1 based on two-dimensional PCR technology / 中华检验医学杂志

Objective:To establish a single-tube, one-step method for detecting and identifying 16 high-risk human papillomavirus (HR-HPV) subtypes (HPV16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68, 73, 82) and genotyping p53 (rs1042522) and RB1 (rs3092905) in cervical cells, using high-throughput two-dimensional PCR (2D-PCR) technology. Methods:Applied Research. Specific primers were designed according to the DNA sequences of the 16 different HR-HPV subtypes, p53, and RB1 genes, with the target genes p53 and RB1 serving as internal references to assess the success of sample collection and PCR amplification. In three fluorescent detection channels, upstream primers labeled with corresponding tags were used for different HR-HPV subtypes, p53, and RB1, constructing a comprehensive 2D-PCR detection system. Using this method, 804 cervical brush samples collected from the gynecology outpatient department of Changzhou First People′s Hospital from December 2022 to August 2023 were tested. The test results were compared for consistency with PCR-reverse dot blot assay, flow cytometric fluorescence hybridization assay, and single-plex real-time quantitative PCR assay, respectively. Meanwhile, the genotypes of p53 and RB1 were detected using Sanger sequencing. The Kappa test was applied to determine the consistency between 2D-PCR method and other methods. Results:2D-PCR accurately discriminated and identified the genotypes of 16 HR-HPV types and p53, RB1 through characteristic melting valleys in the FAM, HEX, and Alexa Fluor568 channels. 2D-PCR showed high consistency with PCR-reverse dot blot assay, with a Kappa value of 0.699, even higher consistency with flow cytometric fluorescence hybridization assay, with a Kappa value of 0.793, and the highest consistency with single-plex quantitative PCR, with a Kappa value of 0.880 (95% CI 0.862-0.907). Using Sanger sequencing as the gold standard, the accuracy of 2D-PCR method in detecting p53 and RB1 genotypes is 100%. The distribution frequencies of the three genotypes (G/G, G/C, and C/C) at the p53 rs1042522 locus were 32.09% (258/804), 49.88% (401/804) and 18.03% (145/804), respectively, while all detected genotypes at the RB1 rs3092905 locus were A/A. Conclusion:This study successfully developed a 2D-PCR method for the identification and genotyping of high-risk human papillomavirus types and related tumor suppressor genes p53 and RB1 for cervical cancer.

The expression of PCDH9 was absent in prostate cancer tissues in the high-score group and was associated with the expressions of p53, Rb and STAT3 / 现代泌尿外科杂志

【Objective】 To explore the expression of PCDH9 loss in regulating cell cycle and promoting tumor progression. 【Methods】 The clinical records of 127 cases of prostate cancer treated during 2018 and 2023 were collected, including 87 paraffin tissue samples from the G4-5 group and 40 from the G1-3 group. The expressions of PCDH9, p53, Rb and STAT3 were detected with immunohistochemical staining, and the relationship between their expressions and clinicopathological characteristics was analyzed. 【Results】 The expression deletion rate of PCDH9 in prostate cancer tissues in G4-5 group (44.8% vs.7.5%) was significantly higher than that in G1-3 group (P<0.001). The positive expression rates of p53 and STAT3 were 34.5% and 89.7%, respectively, and the expression loss rate of Rb was 27.6% in G4-5 group. The expression loss rates of PCDH9 and Rb were associated with neuroendocrine-like histological morphology, nerve invasion and vascular invasion (P<0.05). In G4-5 group of prostate cancer, PCDH9 expression was positively correlated with the expressions of p53 (r=0.345, P<0.05), Rb (r=0.503, P<0.05) and STAT3 (r=0.224, P<0.05). 【Conclusion】 PCDH9 is prone to loss of expression in high-group prostate cancer tissues, especially in cases with neuroendocrine-like histological morphology, which may regulate the cell cycle through the STAT3 signaling pathway, thereby promoting tumor progression.

Mechanism research progress of traditional Chinese medicine intervening in lung cancer by regulating p53 signaling pathway / 中国药房

Lung cancer， the most common malignant tumor， is characterized by a complex pathogenesis and high malignancy， and poses a significant threat to the health and lives of affected individuals. p53 signaling pathway plays a crucial role in the progression of lung cancer and is considered one of the potential targets for targeted therapy. In recent years， multiple studies have indicated that traditional Chinese medicine （TCM） can exert anticancer effects by modulating the p53 signaling pathway. Based on this， this article systematically summarizes the current status and progress of research on TCM intervening in lung cancer by regulating p53 signaling pathway. It was found that TCM formula and preparations， such as Qingjin desheng tablet， Tiaoqi xiaoji decoction， Bufei tongluo jiedu formula， Jianpi bushen formula and Yiqi fuzheng jiedu formula， can promote autophagy and apoptosis of lung cancer cells， inhibit the growth and metastasis of lung cancer cells and strengthen the immune function of the body by activating p53 signaling pathway， thereby inhibiting lung cancer. TCM monomers， such as pseudoginsenoside-Rh2， saikosaponin D， polyphyllin Ⅶ， dendrobiine， sophoridine， gambogic acid， triptolide and triptolide succinate monoester YJ-4， can accelerate cell apoptosis， inhibit the proliferation of lung cancer cells and regulate cell cycle by activating p53 signaling pathway， thereby inhibiting lung cancer.

Mechanism research progress of traditional Chinese medicine intervening in lung cancer by regulating p53 signaling pathway / 中国药房

Lung cancer， the most common malignant tumor， is characterized by a complex pathogenesis and high malignancy， and poses a significant threat to the health and lives of affected individuals. p53 signaling pathway plays a crucial role in the progression of lung cancer and is considered one of the potential targets for targeted therapy. In recent years， multiple studies have indicated that traditional Chinese medicine （TCM） can exert anticancer effects by modulating the p53 signaling pathway. Based on this， this article systematically summarizes the current status and progress of research on TCM intervening in lung cancer by regulating p53 signaling pathway. It was found that TCM formula and preparations， such as Qingjin desheng tablet， Tiaoqi xiaoji decoction， Bufei tongluo jiedu formula， Jianpi bushen formula and Yiqi fuzheng jiedu formula， can promote autophagy and apoptosis of lung cancer cells， inhibit the growth and metastasis of lung cancer cells and strengthen the immune function of the body by activating p53 signaling pathway， thereby inhibiting lung cancer. TCM monomers， such as pseudoginsenoside-Rh2， saikosaponin D， polyphyllin Ⅶ， dendrobiine， sophoridine， gambogic acid， triptolide and triptolide succinate monoester YJ-4， can accelerate cell apoptosis， inhibit the proliferation of lung cancer cells and regulate cell cycle by activating p53 signaling pathway， thereby inhibiting lung cancer.

Effect of Sargassum and Glycyrrhizae Radix et Rhizoma Incompatible Pair with Haizao Yuhutang on Oxidative Stress in Liver of Goiter Rats / 中国实验方剂学杂志

ObjectiveTo observe the effects of Sargassum and Glycyrrhizae Radix et Rhizoma incompatible pair with the Haizao Yuhutang （HYT） on oxidative stress in the liver of goiter rats under the condition of 2 times the dose limit of the Pharmacopoeia of the People's Republic of China 2020. MethodA total of 128 male Wistar rats were randomly divided into a blank group， a model group， a euthyrox group （20 μg·kg-1）， a HYT group （12.06 g·kg-1）， a HYT without Sargassum （HYT-H） group （9.90 g·kg-1）， a HYT without Glycyrrhizae Radix et Rhizoma （HYT-G） group （10.26 g·kg-1）， a HYT without Sargassum and Glycyrrhizae Radix et Rhizoma （HYT-HG） group （8.10 g·kg-1）， and a Sargassum and Glycyrrhizae Radix et Rhizoma （HG） group （3.96 g·kg-1）. The blank group was given deionized water by gavage， and the others were given propylthiouracil （PTU） to replicate the goiter pathological model. Euthyrox was taken as a positive control drug， and the rest of the Chinese medicine groups were given the corresponding decoction by gavage， the material was collected 12 hours after the last dose. The serum levels of aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， alkaline phosphatase （ALP）， and the contents of superoxide dismutase （SOD）， glutathione peroxidase （GSH-Px）， malondialdehyde （MDA）， reactive oxygen species （ROS） in liver tissue were detected in each group. The pathological changes in the liver were observed via hematoxylin-eosin （HE） staining. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was utilized to detect the mRNA expressions of Kelch-like Ech-associated protein 1 （Keap1）， nuclear factor E2-related factor 2 （Nrf2）， heme oxygenase-1 （HO-1）， p53 and Caspase-3 in liver tissues. Western blot was adopted to detect the protein expressions of Nrf2 and HO-1 in liver tissues in oxidative stress-related signaling pathways. ResultCompared with control group， the model group showed significantly increased serum ALT level and contents of MDA and ROS in liver tissues （P<0.05， P<0.01）， significantly reduced activities of SOD and GSH-Px in the liver （P<0.01）， significantly increased mRNA expression of Keap1 （P<0.01）， and significantly decreased mRNA and protein expressions of Nrf2 and HO-1 （P<0.05， P<0.01）. Compared with the model group， the HYT group manifested significantly reduced serum levels of AST， ALT， and ALP （P<0.05， P<0.01）， significantly reduced contents of MDA and ROS in liver tissue （P<0.01）， significantly increased the activities of SOD and GSH-Px （P<0.01）， significantly decreased mRNA expressions of Keap1， p53， and Caspase-3 （P<0.01）， and significantly increased mRNA and protein expressions of Nrf2 and HO-1 （P<0.05， P<0.01）. ConclusionUnder the condition of 2 times the dose limit of the Pharmacopoeia of the People's Republic of China 2020， Sargassum and Glycyrrhizae Radix et Rhizoma incompatible pair with the HYT on oxidative stress in the liver of goiter rats had different effects. The HYT that contains Sargassum and Glycyrrhizae Radix et Rhizoma has a protective effect on the liver of goiter rats， and the effect is better than that of the HG group， the euthyrox group， and the incomplete groups. Its mechanism may be related to activating the Nrf2/HO-1 signaling pathway to alleviate liver oxidative stress and inhibiting the p53/Caspase-3 signaling pathway to reduce hepatocyte apoptosis.

Effects of arsenic and its metabolites on p53 gene expression in BEAS-2B cells / 环境与职业医学

Background Arsenic is a human carcinogen. Arsenic and its metabolites affect the expression of p53, but whether there are any changes of p53 phosphorylation and ubiquitination levels in human bronchial epithelium cells (BEAS-2B) are not clear after exposure to arsenic and its metabolites. Objective To study the effects of arsenic and its metabolites monomethylarsic acid (MMA) and dimethylarsinic acid (DMA) on the expression of tumor suppressor gene p53 in BEAS-2B cells. Methods Different concentrations of sodium arsenite (NaAsO2) were used to infect BEAS-2B cells, and the cell viability was detected with CCK-8 reagent to determine the dose and time of NaAsO2 used for the following study. Based on the results of cell viability, the cells were divided into two panels: a sodium arsenide panel and an arsenic methylation metabolite penal. The doses of sodium arsenite were 0, 2, 4, and 6 μmol·L−1 NaAsO2; the arsenic methylation metabolite panel consisted of 0 μmol·L−1 NaAsO2 group (control), 6 μmol· L−1 MMA group, 6 μmol· L−1 DMA group， and 6 μmol· L−1 NaAsO2 group. The cells were collected after 48 h treatment, and the total protein and total RNA were extracted. The relative levels of p53 mRNA expression were determined by quantitative real-time polymerase chain reaction (qRT-PCR), the relative expression levels of p53 protein, p53 Ser9 and Ser15 phosphorylated proteins were determined by Western blot, and the level of p53 ubiquitination was detected by co-immunoprecipitation (CO-IP). Results Compared with the control group, the cell viability rates in all BEAS-2B cells treated by NaAsO2 were significantly reduced (P<0.05), and the 50% cell viability was observed at 6 μmol·L−1. Compared with the control group, the relative expression level of p53 mRNA gradually decreased after NaAsO2 (2, 4, 6 μmol·L−1) treatment (P<0.05), the relative expression levels of p53 protein and Ser9 phosphorylated protein induced by NaAsO2 also decreased gradually (P<0.05), and the relative expression level of p53 Ser15 phosphorylated protein induced by NaAsO2 followed the same pattern, but it was only lower than that of the control group in the 6 μmol·L−1 NaAsO2 group (P<0.05). Compared with the control group, there were no significant effects on the relative expression levels of p53 mRNA, p53 protein, Ser9 and Ser15 phosphorylated proteins in the MMA group and the DMA group. Compared with the control group, the expression level of p53 ubiquitination was significantly decreased and the expression of K48 ubiquitination decreased significantly after NaAsO2 infection. Conclusion Arsenic causes a decrease in the expression of the p53 protein in BEAS-2B cells, largely due to inhibition of the phosphorylated pathway and a decrease in mRNA expression, and protein changes caused by a decrease in p53 ubiquitination do not play a dominant role. MMA and DMA do not affect p53 gene expression.

Targeting CDT1 inhibit tumor growth and its mechanism in lung adenocarcinoma / 安徽医科大学学报

Objective To explore the clinical significance and mechanisms of chromatin licensing and DNA repli-cation factor 1(CDT1)in lung adenocarcinoma).Methods The gene expression samples of lung adenocarcinoma tissue and normal lung tissue were downloaded from the TCGA database,and perform differential analysis,GO a-nalysis,independent prognosis analysis,and correlation analysis with immunotherapy using R language.CDT1 ex-pression in lung adenocarcinoma and normal tissues was detected by PCR in clinical samples.The changes of cell proliferation and cycle in si-CDT1 knockdown group and si-NC control group were detected by flow cytometry.The invasive ability of each group was detected by Transwell.The expressions of CDT1,TPX2 and p53 in each group were detected by Western blot.Results The TCGA data analysis revealed CDT1 as a differentially expressed gene.GO analysis indicated that CDT1 was closely associated with the cell cycle.The high expression of CDT1 in lung adenocarcinoma tissues was validated in clinical samples.CDT1 could serve as an independent factor for predicting the prognosis of lung adenocarcinoma and had predictive value for immunotherapy in lung adenocarcinoma.Knock-down of CDT1 resulted in a significant decrease in cell proliferation ability compared to the control group,and cells were noticeably arrested in the G1 phase.Transwell assay results demonstrated a significant reduction in invasive capacity in the CDT1 knockdown group.Knockdown of CDT1 led to a significant decrease in TPX2 expression and a significant increase in p53 expression,while overexpression of CDT1 yielded the opposite effect.Conclusion Re-sults demonstrate the elevated expression of CDT1 in lung adenocarcinoma,its association with prognostic signifi-cance,and its impact on lung adenocarcinoma's occurrence and development by influencing TPX2 and p53.

Diagnostic value of plasma SPINK4 expression in colorectal adenocarcinoma and progressive adenoma / 重庆医学

Objective To investigate the clinical diagnostic value of plasma serine protease inhibitor Ka-zal-type 4(SPINK4)expression in colorectal adenocarcinoma(CRC)and progressive adenoma(AA).Methods A total of 62 patients with CRC(CRC group)and 15 patients with AA(AA group)diagnosed by colonoscopy and pathological examination in this hospital from June 2020 to December 2021 were selected,and 22 healthy people undergoing physical examination during the same period were selected as the HC group.The expression of SPINK4 in plasma was detected by ELISA,and the expression of CEA in plasma was detected by electrochemiluminescence,and the correlation was analyzed.The diagnostic efficiency was analyzed by re-ceiver operating characteristic(ROC)curve,and the expression of p53 in CRC tissues was detected by immu-nohistochemistry.Results The expression of plasma SPINK4 in the CRC group and AA group was lower than that in the HC group(Z=3.72,-0.41,P＜0.05),and the expression of CEA in the CRC group was higher than that in the HC group(Z=-3.63,P＜0.05).The area under the curve(AUC),accuracy,sensi-tivity and specificity of SPINK4 combined with CEA in the diagnosis of CRC and AA were higher than those of SPINK4 and CEA alone.The positive rate of mutant type p53 in SPINK4 low expression group and CEA high ex-pression group was significantly increased in CRC patients(72.55％,75.00％,P＜0.05).Conclusion The expression of plasma SPINK4 is decreased in CRC and AA,and the combined detection of SPINK4 and CEA has a good di-agnostic efficiency in CRC and AA.

Effects of microRNA-103a-3p on osteoporosis through tumor protein 53-regulated inhibitor of apoptosis 1/P53 / 解剖学报

Objective To investigate the efeects of microRNA(miR)-103a-3p regulates tumor protein 53-regulated inhibitor of apoptosis 1(TRIAP1)on osteoblast differentiation and bone mass in ovariectomized mice.Methods MC3T3-E1 cells were divided into normal group,miR-103a-3p-NC group,miR-103a-3p mimic group,miR-103a-3p mimic+TRIAP1-NC group,miR-103a-3p mimic+TRIAP1 mimic group.mRNA expression of miR-103a-3p,TRIAP1,P53 were detected by Real-time PCR;Cell proliferation and apoptosis were detected by MTT test and flow cytometry;cytoskeleton and mineralization of cells were detected by F-actin immunofluorescence staining and alizarin staining;alkaline phosphatase(ALP)activity was detected by ELISA.24 female mice were divided into sham group,osteoporosis(OP)group,miR-103a-3p antagonist-NC group,miR-103a-3p antagonist group(six in each group),extract bilateral ovaries to establish an OP model,sham group mice only isolated fat around ovarian tissue.mRNA expression of miR-103a-3p,TRIAP1,P53,ALP,osteocalcin(OCN),osteopontin(OPN)of bone tissue were detected;microCT detect bone mineral density(BMD),bone mineral content(BMC);haematoxylin eosin staining was used to observe pathological changes of bone tissue.Results After miR-103a-3p mimic was transfected into cells,the miR-103a-3p and P53 expression increased,TRIAP1 expression decreased,cell proliferation decreased,apoptosis increased,F-actin expression decreased,the number of calcium nodules decreased,and ALP enzyme activity decreased(P<0.01);however,after TRIAP1 mimic was additionally transfected into cells,the above result caused by miR-103a-3p mimics were significantly reversed(P<0.01).In OP group,the miR-103a-3p and P53 expression in bone tissue increased,the TRIAP1,ALP,OCN and OPN expression decreased,BMD and BMC were decreased,and bone tissue construct was damaged(P<0.05);in miR-103a-3p antagonist group,the miR-103a-3p and P53 expression in bone tissue decreased,TRIAP1,ALP,OCN,OPN expression increased,BMD and BMC increased,and bone tissue construct was improved(P<0.05).Conclusion MiRNA-103a-3p mediate TRIAP1/P53 to inhibit proliferation and mineralization of osteoblast,while miR-103a-3p antagonistic treatment reduce bone loss in OP mice.

Zoledronic acid affects osteogenic differentiation and bone formation by regulating the expression of SIRT3/P53 / 中华内分泌外科杂志

Objective:To explore the mechanism of zoledronic acid (ZOL) affects osteogenic differentiation and bone formation through the regulation of sirtuin 3 (SIRT3) / P53 expression.Methods:Bone marrow mesenchymal stem cells (BMSCs) were induced to differentiate into osteogenic cells, the expression of SIRT3 in the cells was detected, and the targeting regulation relationship between SIRT3 and P53 was analyzed. The intracellular expressions of SIRT3 and P53 were intervened and ZOL was used to treat the cells. MTT method, Western blot method and kit were used to detect cell viability, osteogenesis-related genes Osteoprotegerin (OPG), runt-related transcription factor 2 (Runx2) expression, alkaline phosphatase (ALP) activity and alizarin red S (ARS) staining, respectively. Ovariectomy (OVX) was used to construct a rat model and explore the effect of ZOL on the progression of osteoporosis (OP) in vivo.Results:ZOL promoted osteogenic differentiation of BMSCs. The expression of SIRT3 was down-regulated in the serum of OP patients (0.78±0.23) compared with that of healthy subjects (1.00±0.26 vs. 0.78±0.23. t=3.85, P<0.001). During the osteogenic differentiation of BMSCs, the expression level of SIRT3 gradually increased with the prolonged induction of osteogenesis. Compared with the p53 protein expression and BMSCs activity in the control group, SIRT3 knockout could increase the expression level of p53 protein (0.59±0.05 vs. 1.01±0.11. t=6.02, P=0.004) but inhibited the activity of BMSCs (100.00±8.41 vs. 51.26±5.59. t=8.36, P=0.001). After ZOL treatment, the inhibitory effect of SIRT3 on cell viability (49.61±5.11 vs. 87.61±7.31. t=7.38, P=0.002) and osteogenesis was relieved, and the level of P53 was inhibited (1.10±0.10 vs. 0.69±0.04. t=6.59, P=0.003). P53 overexpression partially offseted the effects of ZOL on cell viability (84.61±6.52 vs. 66.54±5.47. t=3.68, P=0.021) and osteogenesis. Compared with the sham surgery group, the OVX group showed inhibition of osteogenesis in rats, and ZOL treatment significantly improved osteogenic inhibition. ZOL treatment increased the expression level of SIRT3 protein in bone tissue of OVX rats, but inhibited the expression level of P53. Conclusion:ZOL promoted osteogenic differentiation and bone formation of BMSCs by promoting the ubiquitination and degradation of P53 by SIRT3.

Role of cerium oxide nanoparticles in doxorubicin-induced cardiotoxicity and its effect on P53 gene expression / 中国现代医生

@#Objective To study the effect of nano-ceria on doxorubicin-induced cardiotoxic injury and its effect on P53 gene expression,and to explore the mechanism of nano-ceria on doxorubicin-induced cardiotoxic injury.Methods H9C2 myocardial cells were cultured and randomly divided into five groups:control group,model group(1μmol/L adriamycin),nano-cerium oxide group(10μg/ml nano-cerium oxide),experimental group(1μmol/L adriamycin +10μg/ml nano-cerium oxide),and positive control group(1μmol/L adriamycin+10μmol/L dexperimine).The adriamycin induced cardiotoxicity model was established,and the viability of myocardial cells was measured by CCK-8 method.The contents of lactate dehydrogenase(LDH)and malondialdehyde(MDA)in myocardial cells were detected by biochemical method.The levels of reactive oxygen(ROS)and the apoptosis rate in myocardial cells were detected by flow cytometry.The expressions of Bax,Bcl-2 and P53 proteins in myocardial cells were detected by Western blot.Results Compared with the control group,the cell viability was decreased in the model group,the cell LDH and MDA contents were increased,the intracellular ROS level and apoptosis rate were increased,the expressions of Bax and P53 proteins were increased,and the expression of Bcl-2 protein was decreased,and the ratio of Bcl-2/Bax was decreased(all P<0.001).Compared with the model group,the experimental group showed increased cell viability,decreased cell LDH and MDA contents,decreased cell ROS content and apoptosis rate,decreased Bax and P53 protein expressions,and increased Bcl-2 protein expression,and the Bcl-2/Bax ratio was increased(all P<0.001).Conclusion Ceria nanoparticles can effectively prevent adriamycin-induced cardiotoxic injury,and its effect may be related to the down-regulation of P53 gene to inhibit cardiomyocyte apoptosis.

Antitumor Drugs Targeting Mutant p53 Protein / 生物化学与生物物理进展

The p53 protein is an essential tumor suppressor in the human body that plays a critical role in preventing tumor formation by controlling cell cycle arrest and promoting apoptosis. Mutations in the p53 gene are frequently observed in more than 50% of tumor tissues and lead to the generation of mutant p53 proteins, which not only have a dominant-negative effect (DN) that hinders the function of wild-type p53 protein but also have gain-of-function (GOF) effects that stimulate tumor development by regulating cell metabolism, invasion, migration, and other processes. Therefore, mutant p53 protein has become a specific drug target for cancer therapy. However, the lack of a drug-binding pocket and smooth surface of mutant p53 proteins have made them undruggable targets for a long time. In recent years, with the development of high-throughput screening technology and an enhanced understanding of the structure and conformational changes exhibited by mutant p53 proteins, a multitude of small molecule compounds directed against mutant p53 protein have been identified, exhibiting substantial in vitro anti-tumor efficacy. Moreover, some of these compounds have entered clinical trials. This review summarized the direct and indirect strategies for the treatment of cancers targeting mutant p53, with a primary focus on the mechanisms of action of small molecule compounds that reactivate mutant p53 protein or degrade mutant p53 protein. The aim is to provide assistance for the development of innovative drugs targeting mutant p53 protein in the future.