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1.
Article in Chinese | WPRIM | ID: wpr-888186

ABSTRACT

This study explored the effects of resveratrol(Res) on the expression of phosphatase and tensin homolog deleted on chromosome ten(PTEN) and the fibrosis of rat renal tubular epithelial cells in a high-glucose environment and the possible mechanism underlying the fibrosis reduction. After the pretreatment of rat renal tubular epithelial cells(NRK-52 E) cultured in a high-glucose condition with Res or PTEN inhibitor SF1670, they were divided into several groups, i.e., normal glucose(NG), normal glucose + SF1670(NS), high glucose(HG), high glucose + SF1670(HS), high glucose + Res at different concentrations(5, 10, 25 μmol·L~(-1)). The expression and distribution of E-cadherin and α-SMA in renal tubular epithelial cells were observed by immunofluorescence cytochemistry. The protein expression levels of PTEN, E-cadherin, α-SMA, p-Akt~((Thr308)) and collagen Ⅳ were determined by Western blot. Real-time PCR was employed to detect the expression of PTEN mRNA. Compared with the NG group, the HG group witnessed the reduced expression of PTEN mRNA, PTEN protein and E-cadherin protein, but saw the increased expression of α-SMA, p-Akt~((Thr308)) and collagen Ⅳ proteins. Besides, with the increase in Res concentration, the expression levels of PTEN mRNA, PTEN protein and E-cadherin protein gradually increased, while those of α-SMA, collagen Ⅳ, p-Akt~((Thr308)) proteins gradually decreased in the Res groups, showing a dose-effect dependence, compared with the HG group. No distinct difference was found between the NS group and the NG group. The expression level of E-cadherin was even lower and those of α-SMA, p-Akt~((Thr308)), and collagen Ⅳ were higher in the HS group than in the HG group, with no marked difference shown in the two groups in terms of PTEN mRNA and protein. Although the PTEN inhibitor did not affect PTEN, the expression changes of the other proteins were opposite to the results after Res treatment and the fibrosis was aggravated, which suggested that SF1670 promoted the fibrosis by inhibiting PTEN, activating Akt and increasing the synthesis of collagen Ⅳ and other extracellular matrix. The results show that Res can antagonize the high glucose-mediated fibrosis of renal tubular epithelial cells. This may be achieved via the up-regulation of PTEN and the inhibition of PI3 K/Akt signaling pathway.


Subject(s)
Animals , Epithelial Cells , Fibrosis , Glucose , PTEN Phosphohydrolase/genetics , Rats , Resveratrol/pharmacology
2.
Braz. j. med. biol. res ; 54(9): e10390, 2021. graf
Article in English | LILACS | ID: biblio-1249337

ABSTRACT

Sorafenib (SOR) resistance is still a significant challenge for the effective treatment of hepatocellular carcinoma (HCC). The mechanism of sorafenib resistance remains unclear. Several microRNAs (miRNAs) have been identified as playing a role in impairing the sensitivity of tumor cells to treatment. We examined the mechanism behind the role of miR-92b in mediating sorafenib resistance in HCC cells. We detected that miR-92b expression was significantly upregulated in SOR-resistant HepG2/SOR cells compared to parental HepG2/WT cells. After transfection with miR-92b inhibitor, the proliferation of HepG2/SOR cells was remarkably weakened and rates of apoptosis significantly increased. PTEN was considered to be a functional target of miR-92b according to a luciferase reporter assay. Knockdown of PTEN significantly impaired the ability of miR-92b inhibitor on increasing sorafenib sensitivity of HepG2/SOR cells. Furthermore, we confirmed by western blotting and immunofluorescence that miR-92b can mediate sorafenib resistance by activating the PI3K/AKT/mTOR pathway in HCC cells by directly targeting PTEN. These findings further validate the mechanism of miR-92b in SOR resistance in HCC treatment.


Subject(s)
Humans , Carcinoma, Hepatocellular/genetics , Carcinoma, Hepatocellular/drug therapy , Drug Resistance, Neoplasm , MicroRNAs/genetics , Sorafenib/pharmacology , Liver Neoplasms/genetics , Liver Neoplasms/drug therapy , Signal Transduction , Gene Expression Regulation, Neoplastic , Phosphatidylinositol 3-Kinases/metabolism , Cell Line, Tumor , Cell Proliferation , PTEN Phosphohydrolase/genetics , TOR Serine-Threonine Kinases
3.
Braz. j. med. biol. res ; 53(4): e8882, 2020. tab, graf
Article in English | LILACS | ID: biblio-1100927

ABSTRACT

Hepatocellular carcinoma (HCC) is one of the most common primary malignant tumors of the liver worldwide. Liver resection and transplantation are currently the only effective treatments; however, recurrence and metastasis rates are still high. Previous studies have shown that the epithelial-mesenchymal transition (EMT) is a key step in HCC invasion and metastasis. Inhibition of EMT has become a new therapeutic strategy for tumors. Recently, puerarin, a well-characterized component of traditional Chinese medicine, has been isolated from Pueraria radix and exerts positive effects on many diseases, particularly cancers. In this study, CCK-8, EdU immunofluorescence, colony formation, wound healing, and migration assays were used to detect the effects of puerarin on HCC cells. We further analyzed the relationship between puerarin and miR-21/PTEN/EMT markers in HCC cell lines. Our results showed that HCC cell proliferation, migration, invasion, tumor formation, and metastasis were reduced by puerarin in vitro and in vivo. Additionally, puerarin inhibited the EMT process of HCC by affecting the expression of Slug and Snail. Moreover, oncogenic miR-21 was inhibited by puerarin, coupled with an increase in the tumor suppressor gene PTEN. Increasing miR-21 expression or decreasing PTEN expression reversed the inhibition effects of puerarin in HCC. These data confirmed that puerarin affects HCC through the miR-21/PTEN/EMT regulatory axis. Overall, puerarin may represent a chemopreventive and/or chemotherapeutic agent for HCC treatment.


Subject(s)
Animals , Male , Carcinoma, Hepatocellular/drug therapy , PTEN Phosphohydrolase/genetics , Epithelial-Mesenchymal Transition/drug effects , Isoflavones/therapeutic use , Liver Neoplasms/drug therapy , Antineoplastic Agents/therapeutic use , Pyrroles , Drug Screening Assays, Antitumor , Carcinoma, Hepatocellular/genetics , MicroRNAs/genetics , Cell Proliferation/drug effects , Cell Proliferation/genetics , Disease Models, Animal , Isoflavones/pharmacology , Liver Neoplasms/genetics , Neoplasm Invasiveness , Antineoplastic Agents/pharmacology
4.
Braz. j. med. biol. res ; 53(2): e9106, 2020. graf
Article in English | LILACS | ID: biblio-1055491

ABSTRACT

Reperfusion strategies in acute myocardial infarction (AMI) can cause a series of additional clinical damage, defined as myocardial ischemia/reperfusion (I/R) injury, and thus there is a need for effective therapeutic methods to attenuate I/R injury. miR-26a-5p has been proven to be an essential regulator for biological processes in different cell types. Nevertheless, the role of miR-26a-5p in myocardial I/R injury has not yet been reported. We established an I/R injury model in vitro and in vivo. In vitro, we used cardiomyocytes to simulate I/R injury using hypoxia/reoxygenation (H/R) assay. In vivo, we used C57BL/6 mice to construct I/R injury model. The infarct area was examined by TTC staining. The level of miR-26a-5p and PTEN was determined by bioinformatics methods, qRT-PCR, and western blot. In addition, the viability and apoptosis of cardiomyocytes were separately detected by MTT and flow cytometry. The targeting relationship between miR-26a-5p and PTEN was analyzed by the TargetScan website and luciferase reporter assay. I/R and H/R treatment induced myocardial tissue injury and cardiomyocyte apoptosis, respectively. The results showed that miR-26a-5p was down-regulated in myocardial I/R injury. PTEN was found to be a direct target of miR-26a-5p. Furthermore, miR-26a-5p effectively improved viability and inhibited apoptosis in cardiomyocytes upon I/R injury by inhibiting PTEN expression to activate the PI3K/AKT signaling pathway. miR-26a-5p could protect cardiomyocytes against I/R injury by regulating the PTEN/PI3K/AKT pathway, which offers a potential approach for myocardial I/R injury treatment.


Subject(s)
Animals , Rabbits , Myocardial Reperfusion Injury/metabolism , Myocardial Ischemia/metabolism , Phosphatidylinositol 3-Kinases/metabolism , Myocytes, Cardiac/pathology , MicroRNAs/metabolism , PTEN Phosphohydrolase/metabolism , Signal Transduction , Blotting, Western , Disease Models, Animal , Proto-Oncogene Proteins c-akt/metabolism , Real-Time Polymerase Chain Reaction , Flow Cytometry , Mice, Inbred C57BL
5.
Biol. Res ; 53: 01, 2020. graf
Article in English | LILACS | ID: biblio-1089072

ABSTRACT

BACKGROUND: Long non-coding RNA small molecule RNA host gene 1 (SNHG1) was previously identified to be relevant with Parkinson's disease (PD) pathogenesis. This work aims to further elucidate the regulatory networks of SNHG1 involved in PD. Methods: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-hydrochloride (MPTP)-induced mice and 1-methyl-4-phenylpyridinium (MPP+)-treated SH-SY5Y cells were respectively constructed as the in vivo and in vitro PD models. Expression levels of SNHG1 and miR-153-3p were detected by qRT-PCR. Protein expression levels of phosphate and tension homology deleted on chromosome ten (PTEN) were measured by western blotting assay. Cell viability and apoptosis were determined by MTT and flow cytometry assays. The interactions among SNHG1, miR-153-3p and PTEN were identified by luciferase reporter assay, RNA immunoprecipitation, and/or RNA pull-down analysis. RESULTS: Increased SNHG1 expression was found in midbrain of MPTP-induced PD mice and MPP+-treated SH-SY5Y cells. Overexpression of SNHG1 lowered viability and enhanced apoptosis in MPP+-treated SH-SY5Y cells. Moreover, SNHG1 acted as a molecular sponge to inhibit the expression of miR-153-3p. Furthermore, miR-153-3p-mediated suppression of MPP+-induced cytotoxicity was abated following SNHG1 up-regulation. Additionally, PTEN was identified as a direct target of miR-153-3p, and SNHG1 could serve as a competing endogenous RNA (ceRNA) of miR-153-3p to improve the expression of PTEN. Besides, enforced expression of PTEN displayed the similar functions as SNHG1 overexpression in regulating the viability and apoptosis of MPP+-treated SH-SY5Y cells. Finally, SNHG1 was found to activate PTEN/AKT/mTOR signaling pathway in SH-SY5Y cells by targeting miR-153-3p. CONCLUSION: SNHG1 aggravates MPP+-induced cellular toxicity in SH-SY5Y cells by regulating PTEN/AKT/mTOR signaling via sponging miR-153-3p, indicating the potential of SNHG1 as a promising therapeutic target for PD.


Subject(s)
Animals , Male , Mice , Parkinson Disease/metabolism , 1-Methyl-4-phenylpyridinium/toxicity , PTEN Phosphohydrolase/metabolism , Proto-Oncogene Proteins c-akt/metabolism , TOR Serine-Threonine Kinases/metabolism , RNA, Long Noncoding/metabolism , Parkinson Disease/genetics , Transfection , Signal Transduction , Cells, Cultured , Gene Expression Regulation , Blotting, Western , Apoptosis , MicroRNAs , Disease Models, Animal , Real-Time Polymerase Chain Reaction , RNA, Long Noncoding/genetics , Mice, Inbred C57BL
6.
Int. j. morphol ; 37(3): 800-804, Sept. 2019. graf
Article in Spanish | LILACS | ID: biblio-1012356

ABSTRACT

El glioblastoma multiforme es el subtipo de gliomas más frecuente en adultos, con una pobre sobrevida promedio posterior al diagnóstico incluso si se aplica el tratamiento óptimo. Se ha estudiado marcadores tumorales de buen pronóstico, siendo controversial la expresión del homólogo de fosfatasa y tensina. Se estudió muestras parafinadas obtenidas de pacientes con glioblastoma multiforme en el Hospital Carlos Van Buren de Valparaíso, Chile, entre 2010 y 2014. Se realizó análisis inmunohistoquímico para expresión de homólogo de fosfatasa y tensina, estudiándose la intensidad y el patrón de expresión en astrocitos y células epiteliales, además de revisión de datos clínicos. Análisis estadístico utilizando SPSS v20. Se estudió la expresión de PTEN en 21 pacientes. Un 52,4 % presentó una baja expresión en núcleos de astrocitos, con un promedio de sobrevida de 14,2 meses comparado con 10,2 meses del grupo con alta expresión (p=0,33). Se encontró una intensa expresión endotelial en tejido tumoral, comparado con tejido cerebral sin tumor. Se encontró una relación entre la expresión nuclear en astrocitos con diferencias en el tiempo de sobrevida, aunque no estadísticamente significativa, requiriéndose nuevos estudios para corroborarlo. La intensa expresión endotelial observada en tejido tumoral debe ser analizada de forma dirigida.


Glioblastoma multiforme is the most frequent glioma subtype in adults, with poor survival rate after diagnosis even applying the optimal treatment. Tumoural markers have been studied looking for good prognosis, being the phosphatase and tensin homologue controversial. Paraffined samples were used from Carlos Van Buren Hospital in Valparaíso, Chile, between 2010 and 2014. An immunohistochemical analysis was performed looking for phosphatase and tensing homologue expression, studying the intensity and expression pattern in astrocytes and epithelial cells, in addition to clinical data. Statistical analysis was performed using SPSS v20. It was studied the phosphatase and tensin homologue expression in 21 patients. In the study, 52,4 % presented low expression in astrocytic glial cell nuclei, with a survival mean of 14.2 months in comparison to 10.2 months in the high expression group (p=0.33). A very intense endothelial expression was found in tumoural tissue, in comparison to the tissue without tumor. A relation between nuclear expression in astrocytes and survival rate was found, although no statistically significant. The intense endothelial expression seen in tumoural tissue must be studied directly.


Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Young Adult , Brain Neoplasms/metabolism , Glioblastoma/metabolism , Immunohistochemistry , Survival Analysis , Astrocytes/metabolism , Retrospective Studies , PTEN Phosphohydrolase/metabolism
7.
Journal of Experimental Hematology ; (6): 1722-1727, 2019.
Article in Chinese | WPRIM | ID: wpr-781406

ABSTRACT

OBJECTIVE@#To study the PTEN gene expression level and its clinical significance in patients with acute T lymphoblastic leukemia.@*METHODS@#One hundred and twenty-four patients with T-ALL treated in our hospital from January 2014 to May 2018 were selected, and 120 healthy people were selected as control group. The bone marrow of the patients were collected, and mononuclear cells were separated out. PTEN gene expression level was detected by RT-PCR, PTEN protein expression level was detected by Western blot, Kaplan-Meier was used to analyze the survival rate of patients with T-ALL, Cox multivariate regression analyzed was used to analyze the independent influencing factors of patients, and the correlation between PTEN level and clinical characteristics of patients with T-ALL and its prognostic value were analyzed.@*RESULTS@#The relative level of PTEN gene in patients with T-ALL was 0.19±0.06, which was significantly lower than that in control groups (P<0.05). There was significantly positive correlation of the expression level of PTEN gene with white blood cell count (r=0.993)and peripheral blood immature cells (r=0.996) in patients with T-ALL. There was no correlation between PTEN gene expression level and sex, age, LDH level, Hb level, platelet count in patients with T-ALL. And it was found that expression levels PTEN protein in the middle-risk group, the standard-risk group and the high-risk group were significant lower than those in the control group (P<0.05), while those in the high-risk group were significantly lower than those in other groups (P<0.05). Kaplan-Meier survival analysis showed that OS and DFS in PTEN gene high expression group were higher than those in PTEN low expression group (P<0.05). Cox multivariate regression analysis showed that white blood cell count and PTEN gene were independent influencing factors of OS (P<0.05); platelet count and PTEN gene were independent influencing factors of DFS (P<0.05).@*CONCLUSION@#PTEN gene level relates to the prognosis of patients with T-ALL. Patients with better prognosis show a higher PTEN gene level, which provides reference for clinical treatment of patients with T-ALL.


Subject(s)
Bone Marrow , Humans , Leukemia-Lymphoma, Adult T-Cell , PTEN Phosphohydrolase , Prognosis
8.
Journal of Experimental Hematology ; (6): 1548-1555, 2019.
Article in Chinese | WPRIM | ID: wpr-775687

ABSTRACT

OBJECTIVE@#To investigate the biological characteristics of ABCG2 and its effect on side population cells (SP cells) of multiple myeloma (MM) so as to find the way for reversing drug resistance.@*METHODS@#The silence of ABCG2 expression was performed throngh interfering the MM cells by using siRNA. then the ratio of SP cell of MM, speed of cell proliferation and sensitivity of cells to chermotherapentic drugs before and after interference were compared, the biological functions of ABCG2 and its role in regulation of SP cells and PTEN/PI3K/AKT signaling pathway was clarified.@*RESULTS@#siRNA interference could down regulate the expression of ABCG2 at both mRNA and protein level. After siRNA interference, the proliferation of multiple myeloma cells was decreased slightly, the expression of PTEN was increased, the activity of PI3K/AKT pathway was inhibited, and the ratio of SP cells was decreased.@*CONCLUSION@#In multiple myeloma, down-regulation of expression ABCG2 can negatively regulate the expression of PTEN, the PI3K/AKT signaling pathway is inhibited,the ratio of SP decreased,and the response to drug is increased.


Subject(s)
ATP Binding Cassette Transporter, Subfamily G, Member 2 , Cell Line, Tumor , Humans , Multiple Myeloma , Neoplasm Proteins , PTEN Phosphohydrolase , Phosphatidylinositol 3-Kinases , Proto-Oncogene Proteins c-akt , Side-Population Cells , Signal Transduction
9.
Article in English | WPRIM | ID: wpr-763190

ABSTRACT

PURPOSE: Jab1 is a coactivator of c-Jun that enhances the transcriptional function of c-Jun. Jab1 is frequently overexpressed in various cancers and is associatedwith poor prognosis of cancer patients. Thus, Jab1 could be a potential therapeutic target in cancer. However, the role of Jab1 in biliary tract cancer (BTC) has not been studied. MATERIALS AND METHODS: We performed in vitro and in vivo experiments to evaluate the therapeutic potential ofJab1 inhibition in BTC. RESULTS: Among 8 BTC cell lines, many showed higher Jab1 expression levels. In addition, Jab1 silencing by siRNA increased p27 expression levels. SNU478 and HuCCT-1 cells exhibited profound Jab1 knockdown and increased p27 expression by Jab1-specific siRNA transfection. Jab1 silencing induced anti-proliferative and anti-migratory effects and resulted in G1 cell cycle arrest in SNU478 and HuCCT-1 cells. In addition, Jab1 silencing potentiated the anti-proliferative and anti-migratory effects of cisplatin by increasing DNA damage. Interestingly,Jab1 knockdown increased PTEN protein half-life, resulting in increased PTEN expression. In the HuCCT-1 mouse xenograft model, stable knockdown of Jab1 by shRNA also showed anti-proliferative effects in vivo, with decreased Ki-67 expression and AKT phosphorylation and increased Terminal deoxynucleotidyl transferase–mediated dUTP nick end labeling and p27 expression. CONCLUSION: Jab1 knockdown demonstrated anti-proliferative and anti-migratory effects in BTC cells by increasing DNA damage and stabilizing PTEN, resulting in G1 cell cycle arrest. In addition, Jab1 silencing potentiated the anti-proliferative effects of cisplatin. Our data suggest that Jab1 may be a potential therapeutic target in BTC that is worthy of further investigations.


Subject(s)
Animals , Biliary Tract Neoplasms , Biliary Tract , Cell Line , Cisplatin , DNA Damage , G1 Phase Cell Cycle Checkpoints , Half-Life , Heterografts , Humans , In Vitro Techniques , Mice , Phosphorylation , Prognosis , PTEN Phosphohydrolase , RNA, Small Interfering , Transfection
10.
Article in Chinese | WPRIM | ID: wpr-771899

ABSTRACT

OBJECTIVE@#To investigate the effect of Bmi-1 gene silence on the proliferation ability of K562 cells in vitro and in vivo, and to explore the relation of molecular mechanism between proliferation ability of K562 cells in vitro and in vivo with PTEN/pAKT signaling pathway.@*METHODS@#The Bmi-1 small interference RNA (siRNA) sequences were transfected into K562 cells for decreasing Bmi-1 expression. The effect of Bmi-1 siRNA on the proliferation of K562 cells in vitro and in vivo was detected by MTT method and colony-forming test, the effect of Bmi-1 siRNA on the tumorogenicity of K562 cells was observed by subcutaneous inoculation of K562 cells, LY294002 and Bpv treated K562 cells in nude mice, the expression of Bmi-1, PTEN and pAKT proteins were detected by Western blot.@*RESULTS@#The Bmi-1 siRNA could inhibit the proliferation activity, colony-forming and tumor-forming abilities of K562 cells. After the silence of Bmi-1 gene, the PTEN expression in Bmi-1 gene-silenced group was significantly enhanced. While the pAKT expression in Bmi-1 gene-silenced group was significantly reduced; after the K562 cells were treated with LY294002 (an inhibitor of pAKT), the pAKT expression colony-forming and tumor forming abilities were reduced in comparison with untreated K562 cells; after the K562-S1 cells were treated with Bpv (an inhibitor of PTEN), the PTEN expression decreased, while the pAKT expression, colony forming and tumor-forming abilities were restored.@*CONCLUSION@#The Bmi-1 gene possibly involves in regulation of K562 proliferation in vivo and in vitro, the effect of PTEN/pAKT signaling pathway maybe one of molecular mechanisms mediating this regulation.


Subject(s)
Animals , Apoptosis , Cell Proliferation , Humans , K562 Cells , Leukemia , Mice , Mice, Nude , PTEN Phosphohydrolase , Polycomb Repressive Complex 1 , Proto-Oncogene Proteins c-akt , RNA, Small Interfering , Signal Transduction
11.
Journal of Experimental Hematology ; (6): 1053-1057, 2019.
Article in Chinese | WPRIM | ID: wpr-771840

ABSTRACT

OBJECTIVE@#To investigate the relationship of PTEN/PI3K/AKT signaling pathway protein expression with apoptosis and drug-resistance of children's ALL primary cells treated with daunorubicin (DNR).@*METHODS@#The bone marrow mononuclear cells in newly diagnosed and untreated B-ALL children were collected and cultured. After the treatment of primary-cultured cells with DNR of final concentration 0.5 mg/L for 24 h, the cell apoptosis rate was detected by using cell apoptosis assay kit; the samples were collected at the on test of culture and after drug treatment, then expression levels of PTEN, PI3K and AKT proteins were detected by Western blot, moreover the interindex correlation was analyzed.@*RESULTS@#After DNR treatment, the apoptosis rate in PTEN low expression group was lower than that in PTEN high expression group (P<0.05), showing high positive correlation of the cell apoptosis rate with the expression of PTEN before DNR treatment; the cell apoptosis rate in PI3K and AKT low expression group was higher than that in PI3K and AKT high expression group (P<0.01); however, the expression of PI3K and AKT proteins was down-regulated after treatment with DNR (P<0.01).@*CONCLUSION@#The difference of PTEN expression is present in primary cells of B-ALL children, however the change of PTEN expression is not significant after DNR treatment, suggesting that the PTEN expression correlates with DNR-resistance. The DNR can induce the apoptosis of childrens B-ALL primary cells by down-regulating the expression of PI3K and AKT signaling pathway proteins.


Subject(s)
Apoptosis , Child , Daunorubicin , Humans , PTEN Phosphohydrolase , Phosphatidylinositol 3-Kinases , Proto-Oncogene Proteins c-akt , Signal Transduction
12.
Frontiers of Medicine ; (4): 131-137, 2019.
Article in English | WPRIM | ID: wpr-771286

ABSTRACT

The inhibitory environment that surrounds the lesion site and the lack of intrinsic regenerative capacity of the adult mammalian central nervous system (CNS) impede the regrowth of injured axons and thereby the reestablishment of neural circuits required for functional recovery after spinal cord injuries (SCI). To circumvent these barriers, biomaterial scaffolds are applied to bridge the lesion gaps for the regrowing axons to follow, and, often by combining stem cell transplantation, to enable the local environment in the growth-supportive direction. Manipulations, such as the modulation of PTEN/mTOR pathways, can also enhance intrinsic CNS axon regrowth after injury. Given the complex pathophysiology of SCI, combining biomaterial scaffolds and genetic manipulation may provide synergistic effects and promote maximal axonal regrowth. Future directions will primarily focus on the translatability of these approaches and promote therapeutic avenues toward the functional rehabilitation of patients with SCIs.


Subject(s)
Animals , Axons , Physiology , Biocompatible Materials , Genetic Enhancement , Methods , Humans , Nerve Regeneration , PTEN Phosphohydrolase , Metabolism , Recovery of Function , Spinal Cord Injuries , Tissue Engineering , Methods , Tissue Scaffolds
13.
Braz. j. med. biol. res ; 52(12): e8483, 2019. tab, graf
Article in English | LILACS | ID: biblio-1055462

ABSTRACT

PTEN is the most commonly inactivated tumor suppressor gene in primary prostate cancer (PCa) and its loss is associated with poor clinical outcomes. ERG rearrangement is a genomic alteration frequently found in PCa and its prognostic significance has yielded mixed results. Although the association of PTEN and ERG biomarkers has potential impact on clinical outcomes, studies examining the two genes simultaneously are scarce in Brazilian populations. In this study, we retrospectively examined the relationship between ERG expression and PTEN loss in 119 surgically treated prostate cancer patients from Northeastern Brazil through immunohistochemical analysis. ERG expression was found in 41.0% (48/117) of cases and the loss of PTEN detected in 38.1% (40/105) of samples. ERG-positive cases were significantly associated with lower prostate weight; ERG negatively correlated with Gleason score above 6. The lack of associations for PTEN loss alone in this cohort is counter to the literature, which shows that PTEN loss is usually associated with more aggressive disease. The overlapping of the two biomarkers revealed that samples with positive ERG expression without PTEN loss were associated with lower Gleason and lower Grade group. This study contributes with the discussion about the development of the molecular profiling of prostate cancer. The further development of similar studies could help in stratifying specific risk groups, leading to a more personalized therapeutic decision for prostate cancer treatment.


Subject(s)
Humans , Male , Middle Aged , Aged , Aged, 80 and over , Prostatic Neoplasms/metabolism , Gene Expression Regulation, Neoplastic , PTEN Phosphohydrolase/metabolism , Prognosis , Prostatic Neoplasms/surgery , Prostatic Neoplasms/pathology , Immunohistochemistry , Biomarkers, Tumor/genetics , Biomarkers, Tumor/metabolism , Biomarkers, Tumor/blood , Prevalence , Retrospective Studies , Cohort Studies , PTEN Phosphohydrolase/genetics , PTEN Phosphohydrolase/blood , Neoplasm Grading , Transcriptional Regulator ERG/genetics , Transcriptional Regulator ERG/metabolism , Transcriptional Regulator ERG/blood
14.
Braz. j. med. biol. res ; 52(7): e8381, 2019. tab, graf
Article in English | LILACS | ID: biblio-1011592

ABSTRACT

Experiments were conducted to determine if the follicle-stimulating hormone (FSH) receptor binding inhibitor (FRBI) impacts the expression levels of AT-rich interactive domain-containing protein 1A (ARID1A) and phosphatase and tensin homolog (PTEN) in ovaries and blood, as well as expressions of follicle-stimulating hormone cognate receptor (FSHR) gene and proteins. Mice in FRBI-10, FRBI-20, FRBI-30, and FRBI-40 groups were intramuscularly injected with 10, 20, 30, and 40 mg FRBI/kg, respectively, for five consecutive days. Western blotting and qRT-PCR were utilized to determine expression levels of ARID1A and PTEN proteins and mRNAs. Serum ARID1A and PTEN concentrations of the FRBI-40 group were higher than the control group (CG) and FSH group (P<0.05). FSHR mRNA levels of FRBI-20, FRBI-30, and FRBI-40 groups were lower than that of CG and FSH groups on day 15 (P<0.05 or P<0.01). Expression levels of FSHR proteins of FRBI-30 and FRBI-40 groups were lower than those of CG and FSH groups (P<0.05). Levels of ARID1A and PTEN proteins of the FRBI-30 group were greater than CG on days 20 and 30 (P<0.05). FRBI doses had significant positive correlations to levels of ARID1A and PTEN proteins. Additionally, ARID1A and PTEN had negative correlations to FSHR mRNAs and proteins. A high dose of FRBI could promote the expression levels of ARID1A and PTEN proteins in ovarian tissues. FRBI increased serum concentrations of ARID1A and PTEN. However, FRBI depressed expression levels of FSHR mRNAs and proteins in mouse ovaries.


Subject(s)
Animals , Female , Rabbits , Ovarian Neoplasms/metabolism , Receptors, FSH/antagonists & inhibitors , Nuclear Proteins/blood , DNA-Binding Proteins/metabolism , PTEN Phosphohydrolase/blood , Follicle Stimulating Hormone/metabolism , Phosphorylation , Transcription Factors , Nuclear Proteins/metabolism , Transcriptional Activation/genetics , Up-Regulation , Blotting, Western , DNA-Binding Proteins/blood , PTEN Phosphohydrolase/metabolism , Real-Time Polymerase Chain Reaction
15.
Biol. Res ; 52: 26, 2019. graf
Article in English | LILACS | ID: biblio-1011428

ABSTRACT

BACKGROUND: Acute myeloid leukemia (AML) is an aggressive and mostly incurable hematological malignancy with frequent relapses after an initial response to standard chemotherapy. Therefore, novel therapies are urgently required to improve AML clinical outcomes. 4-Amino-2-trifluoromethyl-phenyl retinate (ATPR), a novel all-trans retinoic acid (ATRA) derivative designed and synthesized by our team, has been proven to show biological anti-tumor characteristics in our previous studies. However, its potential effect on leukemia remains unknown. The present research aims to investigate the underlying mechanism of treating leukemia with ATPR in vitro. METHODS: In this study, the AML cell lines NB4 and THP-1 were treated with ATPR. Cell proliferation was analyzed by the CCK-8 assay. Flow cytometry was used to measure the cell cycle distribution and cell differentiation. The expression levels of cell cycle and differentiation-related proteins were detected by western blotting and immunofluorescence staining. The NBT reduction assay was used to detect cell differentiation. RESULTS: ATPR inhibited cell proliferation, induced cell differentiation and arrested the cell cycle at the G0/G1 phase. Moreover, ATPR treatment induced a time-dependent release of reactive oxygen species (ROS). Additionally, the PTEN/PI3K/Akt pathway was downregulated 24 h after ATPR treatment, which might account for the anti-AML effects of ATPR that result from the ROS-mediated regulation of the PTEN/PI3K/AKT signaling pathway. CONCLUSIONS: Our observations could help to develop new drugs targeting the ROS/PTEN/PI3K/Akt pathway for the treatment of AML.


Subject(s)
Humans , Retinoids/pharmacology , Reactive Oxygen Species/metabolism , Antineoplastic Agents/pharmacology , Fluoroimmunoassay , Leukemia, Myeloid, Acute , Signal Transduction , Down-Regulation , Cell Differentiation/drug effects , Cell Survival/drug effects , Phosphatidylinositol 3-Kinases/metabolism , Cell Line, Tumor , Cell Proliferation/drug effects , PTEN Phosphohydrolase/drug effects , PTEN Phosphohydrolase/metabolism , Proto-Oncogene Proteins c-akt/drug effects , Proto-Oncogene Proteins c-akt/metabolism
16.
Appl. cancer res ; 39: 1-6, 2019. ilus, tab
Article in English | LILACS, Inca | ID: biblio-994774

ABSTRACT

Background: PTEN loss is observed in 20­30% of prostate cancers and is associated with a poor outcome, but clinical details of the impact of this biomarker are unclear for intermediate grade tumors. Methods: We investigated 43 radical prostatectomy-derived grade 7 prostate tumors from the Clinics Hospital of Ribeirão Preto. Tissue microarray (TMA) blocks were constructed and PTEN copy number status was determined for all patients through fluorescence in situ hybridization (FISH). To determine the presence of PTEN protein loss in our study cohort, we performed immunohistochemistry (IHC) in TMA sections. We then developed an automated algorithm in HALO™ to identify regions of PTEN protein loss in whole prostate scanned sections from ten patients with known PTEN deletion status by FISH. Clinical analyses were conducted to determine the associations between PTEN loss and patient outcome. All statistical analyses were conducted in R v3.4.3 with P-values below 0.05 being considered statistically significant. Results: In this study of 43 grade 7 tumors, we found PTEN deletions by FISH in 18.9% of tumors, and PTEN protein loss by IHC in 16.3% of tumors. Both techniques were highly concordant and complementary. Clinical analysis demonstrated that PTEN deletion by FISH was significantly associated with positive margin invasion (P = 0.04) and Gleason score upgrade (P = 0.001). Digital image analysis of ten representative tumors demonstrated distinct intratumoral heterogeneity for PTEN protein loss in four tumors. Conclusions: This study shows that PTEN loss in Gleason grade 7 tumors can be heterogeneous and that a systematic analysis of this biomarker using a combination of FISH, IHC, and digital imaging may identify patients with a greater risk of poor outcome (AU)


Subject(s)
Humans , Male , Prostatic Neoplasms/pathology , PTEN Phosphohydrolase/metabolism , Prognosis , Prostatectomy , Prostatic Neoplasms/genetics , Immunohistochemistry , Biomarkers, Tumor , Cohort Studies , In Situ Hybridization, Fluorescence , Genetic Heterogeneity , Neoplasm Grading
17.
J. coloproctol. (Rio J., Impr.) ; 38(1): 1-8, Jan.-Mar. 2018. tab, graf
Article in English | LILACS | ID: biblio-894029

ABSTRACT

ABSTRACT Objectives: Determine immunohistochemical expression of Phosphatase and tensin homolog (PTEN), Phosphatidylinositol 3 kinase (PI3K), Cycloxygenase-2 (COX2) and one proliferation marker (Ki67) in colorectal polyps and correlate with clinical and pathological data in search of carcinogenic pathways. Methods: The reports of 297 polyps diagnosed through endoscopy were reviewed for parameters including age, gender, prior colorectal cancer, the presence of multiple polyps, and polyps' location, appearance and size. Was conducted a microscopic morphometric computerized analysis of immunohistochemical expression using, the selected antibodies and correlated with clinical and pathological variables. Results: The tissue immunohistochemical expression was higher in right colon polyps for the proliferation marker and Phosphatidylinositol 3 kinase (p ≤ 0.0001 and 0.057 respectively). Cycloxygenase-2 and Phosphatase and tensin homolog demonstrated higher tissue immunoexpression in pedunculated polyps (p = 0.009 and 0.002 respectively). Cycloxygenase-2 exhibited higher immunoexpression in larger polyps (p = 0.005). Phosphatidylinositol 3 kinase, Cycloxygenase-2, Phosphatase and tensin homolog and the proliferation marker exhibited higher immunoexpression in high-grade dysplastic polyps (p = 0.031, 0.013, 0.044 and <0.001 respectively). Phosphatase and tensin homolog labeling was higher in polyps with high-grade dysplasia and lower in some of serrated lesions (p = 0.044). Conclusions: The greater expression of the proliferation marker and Phosphatidylinositol 3 kinase in the right colon may be related to right-sided colorectal carcinogenesis. The proliferation marker, Cycloxygenase-2 and Phosphatidylinositol 3 kinase results can be associated with progression of polyps to colorectal cancer. The higher Phosphatase and tensin homolog expression suggests its attempt to control the cell cycle.


RESUMO Objetivos: Determinar a expressão imuno-histoquímica de Fosfatase homóloga a tensina (PTEN), Fosfatidilinositol-3-cinase (PI3K), Ciclooxigenase-2 (COX2) e um marcador de proliferação (Ki67) em pólipos colorretais e correlacionar com dados clínicos e patológicos buscando sua correspondência na carcinogênese. Métodos: Revisados 297 pólipos diagnosticados através de endoscopia quanto a idade, gênero, história de câncer colorretal, número, localização, aparência e tamanho dos pólipos. Realizadas as avaliações morfométricas computadorizadas das expressões imuno-histoquímicas dos marcadores selecionados, que foram correlacionadas com variáveis clínicas e patológicas. Resultados: A expressão do marcador de proliferação e da Fosfatidilinositol-3-cinase foi maior nos pólipos do cólon direito (p = <0,0001 e 0.057 respectivamente). Ciclooxigenase-2 e Fosfatase homóloga a tensina demonstraram maior imunoexpressão em pólipos pediculados (p = 0,009 e 0,002, respectivamente). Ciclooxigenase-2 expressou mais em pólipos maiores (p = 0,005). Fosfatidilinositol-3-cinase, Ciclooxigenase-2, Fosfatase homóloga a tensina e o marcador de proliferação expressaram mais em pólipos com displasia de alto grau (p = 0,031, 0,013, 0,044 e <0,001, respectivamente). Fosfatase homóloga a tensina marcou mais pólipos com displasia de alto grau que lesões serrilhadas (p = 0,044). Conclusões: A maior expressão do marcador de proliferação e Fosfatidilinositol-3-cinase à direita pode estar relacionada à carcinogênese do lado direito do cólon. Os resultados do marcador de proliferação, Ciclooxigenase-2 e Fosfatidilinositol-3-cinase podem ser associados à progressão dos pólipos para câncer. A expressão aumentada de Fosfatase homóloga a tensina sugere tentativa de controle do ciclo celular.


Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Aged, 80 and over , Colorectal Neoplasms/diagnosis , Colonic Polyps/pathology , Ki-67 Antigen/immunology , PTEN Phosphohydrolase/immunology , Cyclooxygenase 2/immunology , Phosphatidylinositol 3-Kinase/immunology
18.
Cancer Research and Treatment ; : 1396-1417, 2018.
Article in English | WPRIM | ID: wpr-717513

ABSTRACT

PURPOSE: This study aimed to probe into the associations among circular RNA ZFR (circ-ZFR), miR-130a/miR-107, and PTEN, and to investigate the regulatory mechanism of circ-ZFR–miR-130a/miR-107–PTEN axis in gastric cancer (GC). MATERIALS AND METHODS: GSE89143 microarray data used in the study were acquired from publicly available Gene Expression Omnibus database to identify differentially expressed circular RNAs inGC tissues. The expressions of circ-ZFR, miR-130a, miR-107, and PTEN were examined by real-time reverse transcription polymerase chain reaction, while PTEN protein expression was measured by western blot. The variation of GC cell proliferation and apoptosis was confirmed by cell counting kit-8 assay and flow cytometry analysis. The targeted relationships among circZFR, miR-130a/miR-107, and PTEN were predicted via bioinformatics analysis and demonstrated by dual-luciferase reporter assay and RNA immunoprecipitation assay. The impact of ZFR on gastric tumor was further verified in xenograft mice model experiment. RESULTS: Circ-ZFR and PTEN were low-expressed whereas miR-107 and miR-130a were highexpressed in GC tissues and cells. There existed targeted relationships and interactions between miR-130a/miR-107 and ZFR/PTEN. Circ-ZFR inhibited GC cell propagation, cell cycle and promoted apoptosis by sponging miR-107/miR-130a, while miR-107/miR-130a promoted GC cell propagation and impeded apoptosis through targeting PTEN. Circ-ZFR inhibited cell proliferation and facilitated apoptosis in GC by sponging miR-130a/miR-107 and modulating PTEN. Circ-ZFR curbed GC tumor growth and affected p53 protein expression in vivo. CONCLUSION: Circ-ZFR restrained GC cell proliferation, induced cell cycle arrest and promoted apoptosis by sponging miR-130a/miR-107 and regulating PTEN.


Subject(s)
Animals , Apoptosis , Blotting, Western , Cell Count , Cell Cycle , Cell Cycle Checkpoints , Cell Proliferation , Computational Biology , Flow Cytometry , Gene Expression , Heterografts , Immunoprecipitation , Mice , Polymerase Chain Reaction , PTEN Phosphohydrolase , Reverse Transcription , RNA , Stomach Neoplasms
19.
Article in Chinese | WPRIM | ID: wpr-775342

ABSTRACT

The aim of this paper was to observe the effect of gambogenic acid on angiogenesis of lung cancer and its preliminary mechanism. After culturing lung adenocarcinoma A549 cells, the conditioned medium was treated with gambogenic acid and then used to culture human umbilical vein endothelial cells (HUVECs) to establish the indirect contact cell co-culture system. A two-dimensional culture model of HUVEC was established with matrigel to observe the effect of gambogenic acid on angiogenesis. DAPI staining was used to observe the morphological changes in HUVEC cells after treatment with gambogenic acid under the fluorescence microscope. Annexin V-FITC/PI staining and flow cytometry analysis were used to determine gambogenic acid's effect on HUVEC cell apoptosis rate. The protein expressions of PI3K, p-PI3K, Akt, p-Akt were measured by Western blot. PTEN-siRNA was transfected into cells, and RT-PCR was used to detect the expression levels of PI3K and Akt genes. Gambogenic acid can significantly inhibit angiogenesis, and its inhibitory effect was dose-dependent. DAPI staining showed apoptotic morphological features of HUVEC cells under fluorescence microscope. Annexin V-FITC/PI staining showed that gambogenic acid induced apoptosis in HUVECs. The results of Western blot showed that the expressions of p-PI3K and p-Akt protein were down-regulated with gambogenic acid, while the expressions of PI3K and Akt protein was insignificant. The results of RT-PCR indicated that the expressions of PI3K and Akt protein were up-regulated by PTEN siRNA. Gambogenic acid can inhibit angiogenesis in lung cancer in vitro, and the mechanism of inhibiting angiogenesis may be related to the PI3K/Akt signaling pathway.


Subject(s)
A549 Cells , Apoptosis , Coculture Techniques , Human Umbilical Vein Endothelial Cells , Humans , Lung Neoplasms , Drug Therapy , Pathology , Neovascularization, Pathologic , Pathology , PTEN Phosphohydrolase , Genetics , Phosphatidylinositol 3-Kinases , Metabolism , Proto-Oncogene Proteins c-akt , Metabolism , Transfection , Xanthenes , Pharmacology
20.
Article in English | WPRIM | ID: wpr-690672

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the relationship between plasma miR-93-5p and the risk of esophageal cancer, as well as the influence of miR-93-5p on the biological function of esophageal cancer cells, exerted through exosomes.</p><p><b>METHODS</b>The expression of plasma miR-93-5p in esophageal cancer patients and healthy controls was analysed by real-time quantitative PCR. The influence of miR-93-5p on the risk and prognosis of esophageal carcinoma was analyzed by conditional logistic regression and survival analysis. The effect of miR-93-5p on the biological function of recipient cells was investigated by establishing an in vitro donor cell co-culture model. The target gene of miR-93-5p was validated by luciferase reporter assay and Western Blotting.</p><p><b>RESULTS</b>Upregulation of plasma miR-93-5p expression significantly increases the risk of esophageal cancer and is associated with poor prognosis. miR-93-5p transferred by exosomes promotes the proliferation of recipient esophageal cancer cells and affects the expression of PTEN and its downstream proteins p21 and cyclin D1.</p><p><b>CONCLUSION</b>Our study provides a reference for the identification of biomarkers for the diagnosis and prognosis of esophageal cancer.</p>


Subject(s)
Aged , Cell Communication , China , Esophageal Neoplasms , Exosomes , Physiology , Female , Humans , Male , MicroRNAs , Metabolism , Middle Aged , PTEN Phosphohydrolase , Genetics , Metabolism , Risk
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