Role of cyclin D1 in glial tumors—A retrospective and observational study

Aim: High-grade glial tumors remain as one of the most lethal malignancies. Cyclin D1 is expressed in some human malignancies and is the potential target of intervention. The present study aims to determine the relationship of cyclin D1 expression with other clinicopathological parameters. Materials and Methods: A cross-sectional study was carried out in a tertiary care center. Biopsy proven 66 cases of glial tumor patients were included in the study. The patients with incomplete clinical details were excluded from the study. Immunohistochemistry using antibodies for IDH 1 and cyclin d1 was done in all the cases. Glial tumors were reclassified according to WHO 2016 classification. Data analysis was performed using SPSS 26.0 for the windows. Result: Among 66 patients, 49 (74.3%) were males and 17 (25.7%) were females. The age of the patients ranged from 20 years to 70 years. Overall, 6.02% were of grade I Glial tumors, 22.7% were of grade II Glial tumors, 19.6% patients were of grade III Glial tumors, and 51.6% patients were of grade IV Glial tumors. Of 66 samples tested cyclin D1 was positive in 25 (37.87%) as high expressers and 7 (10.60%) were low expressers. Our study showed a significant correlation between the expression of cyclin D1 with grade and IDH mutation status, No significant correlation of cyclin D1 was noted with age or sex of the patient. Conclusion: Cyclin D1 was associated with a higher grade of the glial tumor. It can be a potential marker both for prognosis and treatment of glial tumors.

Effect of cyclin D1 on HBV transcription and replication / 临床肝胆病杂志

Objective To investigate the effect of cyclin D1 (with CCND1 as the gene name) on HBV replication and its potential mechanism. Methods With reference to GSE84044 dataset, the Spearman's rank correlation analysis was used to investigate the correlation between the expression levels of genes in liver tissue and serum HBV DNA load in patients with HBV-related liver fibrosis. Cyclin D1 and cyclin D1 T286A mutant were transiently expressed in the HBV cell replication model, and time-resolved immunofluorescence and quantitative real-time PCR were used to measure the levels of HBsAg/HBeAg and HBV DNA in cell culture supernatant; Western blot was used to measure the level of HBV core protein in cells; reverse-transcription quantitative real-time PCR was used to measure the level of HBV RNA in cells; dual-luciferase reporter assay was used to observe the effect of cyclin D1 on the activity of HBV basic core promoter (BCP). GSE83148 dataset was used to investigate the correlation between CCND1 and HBV-related regulatory factors. The independent samples t -test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups. Results The analysis of GSE84044 data showed that 7 cell cycle genes were significantly negatively correlated with HBV DNA load in liver tissue of the patients with HBV-related liver fibrosis (all r < -0.3, all P < 0.05), which included the CCND1 gene ( r =-0.474, P < 0.001). Exogenous expression of cyclin D1 and cyclin D1 T286A mutant reduced the levels of HBsAg, HBeAg, and HBV DNA in culture supernatant of the HBV replication cell model, as well as the levels of HBV core protein and HBV RNA in cells. Exogenous expression of cyclin D1 significantly inhibited the transcriptional activity of HBV BCP. The expression level of CCND1 in liver tissue of chronic hepatitis B patients was significantly positively correlated with the expression of APOBEC3G ( r =0.575, P < 0.001), SMC5 ( r =0.341, P < 0.001), and FOXM1 ( r =0.333, P < 0.001) which inhibited HBV replication, while it was significantly negatively correlated with the expression of the HBV entry receptor NTCP ( r =-0.511, P < 0.001) and HNF1α as the transcription factor for positive regulation of HBV replication ( r =-0.430, P < 0.001). Overexpression of cyclin D1 in HepG2 cells reduced the transcriptional levels of HNF1α and NTCP. Conclusion Cyclin D1 inhibits HBV transcription and replication possibly by downregulating the expression of HNF1α and NTCP.

YAP regulates the liver size during the fasting-refeeding transition in mice

Liver is the central hub regulating energy metabolism during feeding-fasting transition. Evidence suggests that fasting and refeeding induce dynamic changes in liver size, but the underlying mechanisms remain unclear. Yes-associated protein (YAP) is a key regulator of organ size. This study aims to explore the role of YAP in fasting- and refeeding-induced changes in liver size. Here, fasting significantly reduced liver size, which was recovered to the normal level after refeeding. Moreover, hepatocyte size was decreased and hepatocyte proliferation was inhibited after fasting. Conversely, refeeding promoted hepatocyte enlargement and proliferation compared to fasted state. Mechanistically, fasting or refeeding regulated the expression of YAP and its downstream targets, as well as the proliferation-related protein cyclin D1 (CCND1). Furthermore, fasting significantly reduced the liver size in AAV-control mice, which was mitigated in AAV Yap (5SA) mice. Yap overexpression also prevented the effect of fasting on hepatocyte size and proliferation. Besides, the recovery of liver size after refeeding was delayed in AAV Yap shRNA mice. Yap knockdown attenuated refeeding-induced hepatocyte enlargement and proliferation. In summary, this study demonstrated that YAP plays an important role in dynamic changes of liver size during fasting-refeeding transition, which provides new evidence for YAP in regulating liver size under energy stress.

Paraganglioma del cuerpo carotídeo asociado a un adenoma paratiroideo: reporte de caso

Introducción: Los paragangliomas son tumores raros, y más raros aun cuando se presentan con otros tumores endocrinos. Objetivo: Presentar el caso de un paraganglioma del cuerpo carotídeo asociado con adenomas paratiroideos. Materiales y Método: Se presenta la evaluación clínica, imagenológica y fotográfica del caso. Resultados: Se presenta el caso de una paciente de 34 años con una masa cervical en región cervical de un año de evolución y que, durante los estudios de extensión, se encontró que correspondía a un paraganglioma en la bifurcación carotídea izquierda, asociada además con dos adenomas paratiroideos, que fueron resecados sin complicaciones. Discusión: Se discute la fisiopatología, el diagnóstico y manejo en relación con el caso presentado. Conclusión: La presentación de paragangliomas del cuerpo carotídeo asociadas con adenomas paratiroideos es rara, y su evaluación clínica deberá ser individualizada, dado que, si bien el manejo será en su mayoría quirúrgico, el abordaje dependerá de cada caso.

Introduction: Paragangliomas are rare tumors, and they are even rarer when they present with other endocrine tumors. Aim: To present a clinical case of a carotid body paraganglioma associated with parathyroid adenomas. Materials and Method: There are shown the clinical evaluation, images, and photos of the case. Results: We present the case of a 34 years old female patient with a cervical mass, which has grown for a year, and, after extension studies, it was found that the mass corresponded to a paraganglioma located in the left carotid bifurcation, and it was associated with two parathyroid adenomas, all the neoplasms were resected with no complications. Discussion: It is discussed physiopathology, diagnosis and management based on the presented case Conclusion: Carotid body paragangliomas associated with parathyroid adenomas are rare, and the clinical evaluation must be individual, given that, most of the management is surgical, however, the approach will depend on each case.

p53 & Cyclin D1 expression in surgically resected clear margins of oral squamous cell carcinoma

Oral squamous cell carcinoma (OSCC) is one of the most well-known malignancies that affect the human population worldwide. The early diagnosis and early intervention of OSCC help improve the survival rate of the patients. The tumour free surgical margins are a positive prognostic factor for recurrence-free survival. The molecular markers can be used to detect the tumour free surgical margins. Aim: The aim of the study is to evaluate the expression of p53 & Cyclin D1 marker in resected surgical apparently clear margins and to correlate the p53 & Cyclin D1 expression with clinicopathological characteristics and patient outcome. Methods: The study population included retrospective cases of OSCC with apparently clear margins (2017-18) n=10 and Clinicopathological variables relevant to survival analysis were recorded. Finally, two margins were selected from each case, a total of 20 margins were included in this study. Paraffin-embedded wax blocks retrieved and tissue sections were made. Expression of cyclin D1 and p 53 was assessed by the immunohistochemical staining procedure Results: Positive expressions Cyclin D1 in 40% of mild dysplasia margins and 60% in clearance adequate margins were present. p53 expression was seen in 16% of mild dysplasia margins and 84% in clearance adequate margins. The expression of p53 and Cyclin D1 molecular markers are noted in the basal & parabasal layer of epithelium. Conclusion: Molecular markers could play a more reliable method for the assessment of dysplasia at the margins

Application of β-catenin, Cyclin D1 and DKK1 in pathologically aided diagnosis of breast cancer / 国际肿瘤学杂志

Objective:To detect the expression differences of Wnt signaling pathway related molecules β-catenin, Cyclin D1 and Dickkopf-1 (DKK1) in breast disease tissues, and to explore their application value in pathologically aided diagnosis of breast cancer.Methods:From January 2008 to August 2019, 90 cases of breast tissue specimens in the Cancer Center of Guangzhou Medical University were collected, including 30 cases of breast hyperplasia, 30 cases of breast intraductal carcinoma and 30 cases of breast invasive ductal carcinoma. The expressions of β-catenin, Cyclin D1 and DKK1 in breast tissue of each group were detected by immunohistochemistry. Oncomine database and KM plotter database were used to analyze the expression differences of β-catenin, Cyclin D1 and DKK1 in breast cancer and normal breast tissues and their relationships with survival prognosis of patients with breast cancer, and to verify the results of immunohistochemistry. Receiver operating characteristic (ROC) curve was used to evaluate the efficacies of each molecule in pathologically aided diagnosis.Results:There were statistically significant differences in β-catenin, Cyclin D1 and DKK1 expressions among breast hyperplasia, breast intraductal carcinoma and breast invasive ductal carcinoma ( χ2=7.766, P=0.021; χ2=24.133, P<0.001; χ2=11.585, P=0.003). The expression of β-catenin in breast invasive ductal carcinoma group was significantly higher than that in breast intraductal carcinoma group and breast hyperplasia group ( Z=-2.367, P=0.018; Z=-2.462, P=0.014). The expression of Cyclin D1 in breast invasive ductal carcinoma group and breast intraductal carcinoma group was significantly higher than that in breast hyperplasia group ( Z=-4.166, P<0.001; Z=-4.174, P<0.001). The expression of DKK1 in breast invasive ductal carcinoma group and breast intraductal carcinoma group was significantly higher than that in breast hyperplasia group ( Z=-3.090, P=0.002; Z=-2.923, P=0.003). The results of bioinformatics analysis showed that compared with normal breast tissue, the expression of β-catenin mRNA in invasive breast cancer tissue increased by 2.33 times ( t=15.242, P<0.001), the expression of Cyclin D1 mRNA in breast intraductal carcinoma tissue increased by 6.64 times ( t=7.152, P=0.006), while the expression of DKK1 mRNA in normal breat tissue was 3.41 times higher than that in invasive breast cancer tissue, with no statistically significant difference ( t=-13.193, P>0.999). The median survival time of breast cancer patients in Cyclin D1 high expression group was 173.2 months, which was shorter than 228.9 months in low expression group ( P<0.001). The upper quartile survival time of breast cancer patients in DKK1 high expression group was 55.1 months, which was longer than 40.4 months in low expression group ( P<0.001). The breast invasive ductal carcinoma and breast intraductal carcinoma were combined into tumor group, the sum of the immunohistochemistry scores of β-catenin and Cyclin D1 minus the immunohistochemistry score of DKK1 was used as the combined scoring scheme 1, and the sum of β-catenin and Cyclin D1 immunohistochemistry score was used as the combined scoring scheme 2. ROC curve analysis showed that the area under the curve (AUC) of β-catenin, Cyclin D1, combined scoring scheme 1 and combined scoring scheme 2 for pathologically aided diagnosis of breast cancer were 0.65 ( P=0.080), 0.81 ( P<0.001), 0.70 ( P=0.023) and 0.78 ( P=0.001), respectively. The AUC of Cyclin D1 and combined scoring scheme 2 were ≥0.7, which had good value in pathologically aided diagnosis. Conclusion:Wnt signaling pathway related molecules Cyclin D1 and Cyclin D1 combined with β-catenin detection has a good value in the pathologically aided diagnosis of breast cancer.

Regulatory effects of ADT-OH on the proliferation of neural precursor cells / 中国组织工程研究

BACKGROUND: Researchers believe that hydrogen sulfide (H2S), as an important cell protective molecule, may become a new treatment method to restore the physiological function of diseased cells or organ systems through the artificial regulation of endogenous H2S biosynthesis or in vitro administration of H2S donor. ADT-OH is a slow-release donor of H2S that can improve the survival rate of hippocampal nerve cells with glutamate-induced injury, but studies on the proliferation of cerebral cortical neural precursor cells are rare. OBJECTIVE: To investigate the effect of ADT-OH on the proliferation of neural precursor cells in embryonic cerebral cortex. METHODS: Neural precursor cells from cerebral cortical ventricular zone and subventricular zone of embryonic mice at embryonic 14.5 days were isolated. Neural precursor cells from one fetal mouse were inoculated into one well (24-well plate), and cultured with the medium containing 100 μmol/L ADT-OH. The size and number of neural spheres per well were measured at 3 days after culture. The proliferation rate of cultured neural precursor cells was detected by BrdU labeling. The proliferation of the cells was further verified by immunofluorescence staining with the specific antibody Ki67. The expression of cyclin D1 was finally detected by western blot assay. RESULTS AND CONCLUSION: Our experimental results showed that ADT-OH could promote the formation of neural spheres, and further detection by BrdU and Ki67 antibody showed that ADT-OH could promote the proliferation rate of neural precursor cells. Meanwhile, the expression of cyclin D1, a proliferation-related gene, was up-regulated in neural precursor cells after ADT-OH treatment. Overall, ADT-OH may promote the proliferation of neural precursor cells by regulating the expression of cyclin D1.

miR-361-5p reverses oxaliplatin resistance of gastric cancer SGC-7901 cells by targeting CCND1 / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effects of miR-361-5p on the oxaliplatin (OXA) resistance of gastric cancer SGC-7901 cells and its mechanism. Methods: The expression of miR-361-5p in gastric cancer cells (MKN-45, MGC80-3 and SGC-7901) and drug-resistant SGC-7901/OXA cells was detected by qPCR. The SGC-7901/OXA cells were transfected with miR-361-5p mimics/inhibitor or sh-CCND1 by using Liposome transfection technology. Then, cell proliferation, apoptosis and cell cycle of SGC-7901/OXA cells were measured by CCK-8 assay and Flow cytometry, respectively. The targeting relationship between miR-361-5p and CCND1 was examined by Dual luciferase report gene assay. The expression level of CCND1 in SGC-7901/OXA cells was detected by WB. Results: miR-361-5p was down-regulated in multiple gastric cancer cells and SGC-7901/OXA cells (P<0.05 or P<0.01). Over-expression of miR-361-5p significantly promoted the apoptosis, induced G0/G1 cell cycle arrest and inhibited cell proliferation of SGC-7901/OXA cells (P<0.05 or P<0.01). Dual luciferase reporter gene results verified that miR-361-5p targeted CCND1 and negatively regulated its expression (P<0.01). Further experiments showed that targeted down-regulation of CCND1 induced apoptosis and G0/G1 cell cycle arrest and inhibited CCND1 expression and proliferation of SGC-7901/OXA cells (P<0.05 or P<0.01). Over-expression of miR-361-5p targetedly down-regulated CCND1 and further promoted cell apoptosis, induced G0/G1 cell cycle arrest and inhibited cell proliferation of SGC-7901/OXA cells (P<0.05 or P<0.01). Conclusion: miR-361-5p over-expression can reverse the resistance of SGC-7901/OXA cells to OXA, and the mechanism may be related to its targeted down-regulation of CCND1 expression.

Association of different patterns of expression of beta-catenin and cyclin D1 with pathogenesis of breast carcinoma

Background: Beta-catenin and cyclin D1 have attracted considerable attention in recent studies as potential proto-oncogenes in many human cancers especially colonic cancer. Beta-catenin plays multiple roles within the cell such as canonical Wnt signaling where cyclin D1 has been identified as one of its target genes. The role of beta-catenin and cyclin D1 in breast cancer has been evaluated in many studies but not established yet. Materials and Methods: The expression of beta-catenin and cyclin D1 was evaluated in 82 cases of breast carcinoma (BCa) and 32 cases of ductal carcinoma in situ(DCIS) by immunohistochemistry (IHC). Their relationship with clinicopathological features was also investigated. Statistical analysis was done to establish an association. Results: Abnormal expression of beta-catenin (ABE) was seen in 80.2% cases of invasive ductal carcinoma (IDC) and 47% cases of DCIS, while the cyclin D1 positive expression rate was 60.9% and 50%, respectively. In the cases showing ABE, cyclin D1 positivity was 88.1%. ABE showed significant association with high-grade BCa. The most common pattern of ABE was loss of membrane with nuclear positivity which is associated with worst prognosis. In addition, ABE in cases of BCa and DCIS showed concordant patterns. Conclusion: Therefore, an association exists between ABE and cyclin D1 in BCa and its precursor lesions implying that Wnt/beta-catenin oncogenic pathway may have a definite role in breast carcinogenesis and can be used for targeted therapy. Also, different patterns of beta-catenin expression may have prognostic and predictive value.

Expression pattern research of Cyclin D1 in scored human lens epithelial cells cultured with high glucose / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To observe the expression pattern of Cyclin D1 in human lens epithelial cells(HLECs)after traumatic stimulation in high-glucose culture<p>METHODS: The activity of HLECs was detected by MTT method after incubate with differernt concentration glucose for 24h <i>in vitro </i>to determine the optimal glucose concentration. qRT-PCR and Western blot were used to detect the high glucose pretreatment group and the non-high glucose pretreatment group. The expression of Cyclin D1 in HLECs at different time points after traumatic stimulation was detected.<p>RESULTS: The viability of HLECs were increased when treatment with low concentration glucose, but the concentration should not exceed 25.5mmol/L, or it will inhibit the activity of HLECs; The reasult of high glucose pretreatment group reveal that the expression of Cyclin D1 is down-regulated in a time-dependent manner within a certain time range. While the expression of Cyclin D1 was irregular in the non-pretreatment group, it was increased at the time point of 12h and 48h. The score treatment can up-regulate the expression of Cyclin D1 in HLECs in a certain degreen.<p>CONCLUSION: The effects of glucose on HLECs activity and Cyclin D1 experssion are irrugular. Trauma treatment can stimulate the expression of Cyclin D1 in HLECs to some extent.

Inhibitory effect of celecoxib on Cal-27 tongue squamous cell carcinoma cell proliferation / 口腔疾病防治

Objective@# To explore the inhibitory effect of celecoxib (CELE) on the proliferation of tongue squamous cell carcinoma Cal-27 cells and its mechanism.@*Methods@# A CCK-8 assay was used to investigate the cytotoxicity of different concentrations CELE(10, 20, 40, 60, 80, and 100 mol/L) at 24 and 48 h in Cal-27 cells. According to the concentration of CELE, samples were divided into a control group (0 μmol/L) and experimental groups (10, 20, and 40 μmol/L), and cell invasiveness was detected by the Transwell method. The expression levels of c-Myc and Cyclin D1 mRNA were detected with qPCR, and western blots were used to detect the expression of phosphate and tension homologue deleted on chromosome ten (PTEN), phospho-protein kinase B (p-AKT) (Thr308), c-Myc, cyclin D1 and other proteins in Cal-27 cells after 24 h of treatment with different doses of CELE (10, 20, and 40 μ mol/L) and after 6, 12, and 24 h of treatment with 40 μmol/L CELE.@*Results @# The different concentrations of CELE were able to inhibit the proliferation of Cal-27 cells, and the higher the concentration of CELE was, the more significant the inhibition of the proliferation of Cal-27 cells was. The cell survival rates of cells exposed to 40 μmol/L CELE were 80% and 75% after 24 and 48 h, respectively. In the four groups of patients, the number of invasive cells was compared, and the results in decreasing order were the control group, 10 μmol/L CELE, 20 μmol/L CELE, and 40 μmol/L CELE. The expression level of c-Myc, cyclin D1 mRNA and the protein in P-AKT (Thr308), c-Myc, and cyclin D1 significantly decreased and the expression of PTEN protein increased in the Cal-27 cells after administration of CELE at different concentrations. @*Conclusion@# CELE can inhibit the proliferation of Cal-27 cells, possibly through inhibition of the expression of proliferation signal factors, such as c-Myc and cyclin D1, by activating the PTEN signaling pathway.

lncRNA HOTAIR promotes malignant biological behaviors of breast cancer SKBR3 cells through miR-519d-3p/CCND1 axis / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To explore the effect of lncRNA HOTAIR/miR-519d-3p/cyclin D1 (CCND1) axis on the proliferation and metastasis of breast cancer cells and its underlying mechanism. Methods: A total of 50 pairs of breast cancer tissues and corresponding para-cancer tissues resected from breast cancer patients in the Department of Breast Surgery, the Third Hospital of Nanchang from March 2017 to February 2019 were collected for this study. The expression level of HOTAIR in breast cancer tissues and paired paracancer tissues was detected by qPCR, in addition, the expressions of HOTAIR and miR-519d-3p in normal breast epithelial cells and breast cancer cell lines were also detected. Breast cancer SKBR3 cells were divided into NC group (without any treatment), si-HOTAIR group, mir-519d-3p mimics group, miR-519d-3p mimic+pcHOTAIR group, miR-519d-3p mimic+pcCCND1 group, and si-HOTAIR+ pcCCND1 group. The proliferation ability of SKBR3 cells was detected by CCK-8. Invasion and migration of SKBR3 cells were detected by Transwell. The expression levels of E-cadherin, N-cadherin, Vimentin and CCND1 in SKBR3 cells were detected by Western blotting. The targeting relationship between HOTAIR and miR-519d-3p, miR-519d-3p and CCND1 was detected by Dualluciferase reporter gene system. Results: HOTAIR was highly expressed in breast cancer tissues and cell lines, with the highest expression in SKBR3 cells. HOTAIR knockdown significantly inhibited the proliferation, invasion and migration of SKBR3 cells, as well as increased the expression level of E-cadherin and decreased the expression levels of N-cadherin and Vimentin. Dual-luciferase reporter gene assay showed that HOTAIR targetedly down-regulated the expression of miR-519d-3p, and miR-519d-3p targetedly downregulated the expression of CCND1. Further studies showed that knockout of HOTAIR inhibited the EMT, proliferation, invasion and migration of SKBR3 cells through enhancing the inhibitory effect of miR-519d-3p on CCND1 expression (all P<0.05). Conclusion: HOTAIR knockdown inhibits proliferation and metastasis of SKBR3 cells by regulating the axis of miR-519d-3p/CCND1.

Expression of Cyclin D1 in wound healing of human corneal epithelial cells with high blood glucose level / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To study the effect of high glucose environment on human corneal epithelial cell injury and repair, and to explain the significance of Cyclin D1 protein expression in corneal epithelial cell wound healing in high glucose culture. <p>METHODS: The high-glucose micro-environment of diabetic corneal lesions was simulated. After human corneal epithelial cells were resuscitated, cultured and passaged, a normal control group of DMEM complete medium of equal volume of distilled water and a high-glucose treated group of DMEM complete medium containing 25mmol/L glucose were set respectively. After the cells were overgrown, the cells were stimulated with scratches. The growth conditions and changes of the cells in each group were observed and compared under an inverted phase contrast microscope. Western glucose was used to analyze high glucose at different time points(0, 12, 24, 48, and 72h)Cyclin D1 Protein expression in cultured corneal epithelial cells. The qRT-PCR was used to analyze high glucose at different time points and each group Cyclin D1 mRNA expression.<p>RESULTS: Under the conditions of high glucose treatment <i>in vitro</i>, the repair rate of human corneal epithelial cells was slowed down after injury, floating cells increased, cells reattached less, and cell spacing increased. With the increase of high glucose treatment time, the cell state became worse and the growth rate slow; normal group repaired cell damage faster, increased cell density, regular morphology, and smooth cell membrane. Cyclin D1 expression was up-regulated by Western blot, but the up-regulation effect gradually weakened with time. The highest expression of Cyclin D1 in both groups appeared at 12h. The expression of Cyclin D1 in the high glucose treatment group was lower than that in the normal control group. The qRT-PCR results showed that after high glucose treatment, the expression of Cyclin D1 mRNA was up-regulated, but with the increase of high glucose treatment time, the up-regulation effect weakened, and the mRNA level recovered to the same level as the control group at 48h. <p>CONCLUSION: In the process of corneal epithelial cell wound healing, high glucose negatively regulates and inhibits the expression of Cyclin D1 protein, and is related to the decline of corneal epithelial cell proliferation and apoptosis.

Expression of Cyclin D1 in normal and hyperplastic endometrium

Background: Endometrial hyperplasia is characterised by increased gland to stroma ratio with varying degree of atypia. Cyclin D1 is a protein playing important role during the G1→S phase transition in the cell cycle. The present study evaluated the expression of Cyclin D1 in normal and hyperplastic endometrium.Methods: A cross sectional study was conducted over a period of 1 year. We evaluated and compared the expression of Cyclin D1 in 56 endometrial samples including 24 cases of simple hyperplasia, 12 cases of complex hyperplasia and 10 cases each of secretory and proliferative endometrium.Results: A substantial increase in expression of Cyclin D1 was seen in hyperplastic endometrium compared to normal endometrium. Moreover, complex hyperplasia showed the maximum positivity for Cyclin D1.Conclusions: Cyclin D1 may play a stimulatory role in the proliferation of endometrial glands and hence may be involved in endometrial tumorigenesis.

Anticancer activity of britannin through the downregulation of cyclin D1 and CDK4 in human breast cancer cells

Aim of the Study: Both apoptotic induction and cell cycle blockade in cancer cells are effective strategies to eliminate cancer cells. Many conventional cancer drugs that induce apoptosis and inhibit cell cycle progression have been reported as potential therapeutics for various types of cancer. Britannin is a natural sesquiterpene lactone that its profound anticancer properties were revealed in our previous study. In this study, we evaluated the effects of britannin on the cell cycle distribution and also cell cycle-related proteins. Materials and Methods: Analysis of cell cycle distribution was carried out using flow cytometer. The effects of britannin on cyclin D1 and CDK4 expression were evaluated using the Western blot. Results: The obtained results show that britannin at the low concentrations induces cell growth inhibition mainly through G1-phase arrest while it seems that apoptosis contributes to cell growth inhibitory effect of high doses of britannin. Reduction of cyclin D1 and CDK4 protein levels were also observed after treating cancer cells with britannin. Conclusion: The obtained results reveal that britannin can inhibit MCF-7 and MDA-MB-468 breast cancer cells proliferation through arresting cell cycle progression through cyclin D1/CDK4-mediated pathway

Study of expression of PTEN and Cyclin D1 in endometrium at a tertiary care centre

Background: To study the expression of PTEN (Phosphatase and Tensin homologue) and Cyclin D1 in normal, hyperplastic and neoplastic endometrium by immunohistochemistry and to corroborate the interrelationship between PTEN and Cyclin D1 in normal to neoplastic endometrial disorders including endometrial carcinoma.Methods: Formalin fixed paraffin embedded (FFPE) sections of spectrum of endometrium in fifty different cases were taken from secretory phase to endometrial carcinoma and subjected to Immunohistochemistry using PTEN and Cyclin D1 .Results: Immunoreactivity was regarded as positive when brown staining was localized in the nuclei or cytoplasm. The intensity of nuclear staining was graded from 0 to 3+ and the extent was semi quantitatively estimated. If less than 10% of cells were positive a score of 0 was given, 11 % to 30% cell positivity was scored as 1+, 31% to 60 % positivity was scored as 2+ and more than 60% positive cells was labelled as 3+. Statistical analysis was performed with Chi-Square test and significant differences were noted between these 3 groups (p value < 0.05).Conclusions: The present study supports that an inverse correlation exists in the expression of PTEN and Cyclin D1 in normal, hyperplastic and neoplastic endometrium. The decreased PTEN expression is a marker of the earliest endometrial premalignant lesions, and we propose that use of PTEN immunostaining may be informative in identifying premalignant lesions that are likely to progress to carcinoma. Cyclin D1 expression in endometrial glands increases progressively in intensity and extent from normal endometrium to hyperplasia to carcinoma.

Main active components combination of Astragalus membranaceus and Angelica sinensis promotes proliferation of aging hematopoietic stem cells / 中草药

Objective To explore the effects of the combination of main active components of Astragalus membranaceus and Angelica sinensis such as astragaloside IV, formononetin, calycosin, campanulin, ferulic acid on aging hematopoietic stem cells (HSCs), and clarify its mechanism through cell cycle regulation. Methods The aging model of HSCs in mice was established with three butyl hydrogen peroxide (t-BHP) to research the effects of five active components of different concentrations on the senescence and the proliferation of HSCs, and seek the main active components which could promote cell proliferation. Finally, HSCs aging model was used to prepare the drug-containing plasm of A. membranaceus combined with A. sinensis at 1:1 ratio. Furthermore, blank control group, model group, blank plasma group, ferulic acid group, astragaloside IV group, formononetin group, calycosin group, calycosin glycoside group, combination group of main active components, drug-containing plasma group of A. membranaceus combined with A. sinensis at 1:1 ratio were acted on aging cells, HSCs senescence rate was tested by SA-β-galactosidase staining and cell proliferation rate was measured by CCK-8 method, cell cycle distribution was determined by flow cytometry, and the protein expression of Cyclin D1 and cyclin dependent kinase 4 (CDK4) were detected by Western blotting. Results Ferulic acid, astragaloside IV, and formononetin significantly promoted the proliferation of aging HSCs and decreased the positive rate of senescent cells, but the effects of calycosin and calycosin glycoside on HSCs proliferation and the positive rate of senescent cells were not significant. The orthogonal experiment showed that the combination of five active components that ferulic acid, formononetin, astragaloside IV were taken as basic factors, and calycosin and calycosin glycoside were taken as secondary factors, had the strongest effect on promoting cell proliferation and decreasing the positive rate of senescent cells. Ferulic acid, astragaloside IV, formononetin, active component combination and drug-containing plasma decreased the positive rate of senescent cells, down-regulated G0/G1 phase cells while up-regulated G2/M + S phase cells, and increased the expression of Cyclin D1 and CDK4 proteins. The above effects in the active component combination group and the drug-containing plasma group were the best. Conclusion The main active components of A. membranaceus and A. sinensis such as ferulic acid, astragaloside IV, and formononetin can promote the proliferation and improve the senescent of aging HSCs, however, calycosin and calycosin glycoside have no obvious effect. The effect of promoting the proliferation is the strongest on aging HSCs when five active components are combined, and the combination can improve HSCs senescence, enhance the transformation of HSCs from static stage to proliferative stage. The main active components and the combination of A. membranaceus and A. sinensis can promote HSCs proliferation and antagonize HSCs senescent, which may be related to regulating the expression of cell cycle related proteins and promoting the transformation of cell cycle.

Sargentodoxa cuneata combined with chemotherapy drug 5-fluorouracil inhibits growth of liver cancer cells / 中草药

Objective To investigate the effect of Sargentodoxa cuneata extracts combined with 5-fluorouracil on the growth inhibition of human hepatoma HepG2 cells, and explore its mechanism. Methods HepG2 cells were cultured in vitro and treated with different concentrations of S. cuneata extracts. The effect of S. cuneata extracts on cell growth inhibition was detected by MTT assay, and the subsequent experimental concentration was selected. HepG2 cells were randomly divided into four groups: control group, S. cuneata extracts group (40 mg/L), 5-fluorouracil (10 μmol/L) group, and combination group (S. cuneata extracts 40 mg/L + 5-fluorouracil 10 μmol/L). Cell proliferation inhibition rate was detected by MTT assay and cell cycle was detected by flow cytometry. The expression levels of nuclear antigen PCNA, Cyclin D1, and cell cycle-dependent protein kinase CDK4 were detected by Western blotting. Results The results of MTT assay showed that S. cuneata extracts inhibited the proliferation of HepG2 cells in a concentration-dependent manner (P < 0.05), and S. cuneata extracts with a concentration of 40 mg/L was selected for subsequent experiments. Compared with the control group, the cell proliferation inhibition rate of the S. cuneata extracts group and the 5-fluorouracil group was significantly increased, the proportion of G0/G1 phase cells was significantly increased, and the proportion of cells in the S phase and G2/M phase was significantly decreased (P < 0.05). The protein expression levels of PCNA, Cyclin D1, and CDK4 in the cells were significantly decreased (P < 0.05). Compared with the 5-fluorouracil group, the combination group significantly inhibited the proliferation of HepG2 cells, blocked the cell cycle, and inhibited the protein expression of PCNA, Cyclin D1, and CDK4 in the cells (P < 0.05). Conclusion S. cuneata extracts combined with 5-fluorouracil enhances the growth inhibition of hepatocellular carcinoma HepG2 cells, and its mechanism may be related to the inhibition of the protein expression of PCNA, Cyclin D1, and CDK4 related to the expression levels of PCNA, Cyclin D1, and CDK4 in the inhibited cells.

2,3,7,8-Tetrachlorodibenzo-p-dioxin Promotes Proliferation of Astrocyte Cells via the Akt/STAT3/Cyclin D1 Pathway / 生物医学与环境科学（英文）

OBJECTIVE@#The compound 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), a persistent organic pollutant, is harmful to the nervous system, but its effects on the brain are still unclear. This study aimed to investigate the effects of TCDD on astrocytes proliferation and underlying molecular mechanism.@*METHODS@#The cell proliferation was measured by EdU-based proliferation assay and PI staining by flow cytometry. Protein expression levels were detected by Western blotting. Immunofluorescence, cytoplasmic and nuclear fractions separation were used to assess the distribution of signal transducer and activator of transcription 3 (STAT3).@*RESULTS@#C6 cells treated with 10 and 50 nmol/L TCDD for 24 h showed significant promotion of the proliferation of. The exposure to TCDD resulted in the upregulation in the expression levels of phosphorylated protein kinase B (p-Akt), phosphorylated STAT3, and cyclin D1 in a dose- and time-dependent manner. The inhibition of Akt expression with LY294002 or STAT3 expression with AG490 abolished the TCDD-induced cyclin D1 upregulation and cell proliferation. Furthermore, LY294002 suppressed the activation of STAT3. Finally, TCDD promoted the translocation of STAT3 from the cytoplasm to the nucleus, and LY294002 treatment blocked this effect.@*CONCLUSION@#TCDD exposure promotes the proliferation of astrocyte cells via the Akt/STAT3/cyclin D1 pathway, leading to astrogliosis.

Low Doses of Nonylphenol Promote Growth of Colon Cancer Cells through Activation of ERK1/2 via G Protein‒Coupled Receptor 30 / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Nonylphenol (NP) is an endocrine disruptor found in products such as cleaners, plastics, and detergents. It exerts actions similar to endogenous 17β-estradiol (E2) and is reported to influence various cancers. However, its role in colon cancer remains elusive. MATERIALS AND METHODS: Colon cancer cell lines COLO 205 and SW480 were employed in our study. The cells were treated with NP or E2 followed by measurement of apoptosis and proliferation using flow cytometry and MTT assays, respectively. G protein–coupled estrogen receptor 30 (GPR30) expression was visualized using immunofluorescence and Western blot. To investigate the underlying mechanism, the expression levels of GPR30, p-protein kinase A (PKA), c-myc, cyclin D1, and ERK1/2 were analyzed using Western blot. Meanwhile, the GPR30 antagonist G15 was utilized to validate the role of GPR30 in colon cancer progression. Finally, the effect of a GPR30 inhibitor on tumor growth was determined in vivo using tumor xenograft mouse models. RESULTS: NP facilitated the proliferation of colon cancer cells and induced apoptosis failure in vitro. Western blot revealed increased GPR30 expression levels in response to NP treatment. Cyclin D1, p-PKA, c-myc, and proliferating cell nuclear antigen, proteins that regulate the cell cycle, were all upregulated by NP, and NP-mediated ERK1/2 activation and subsequent cell proliferation were abrogated by the GPR30 inhibitor G15. Moreover, colon cancer mice that received G15 administration demonstrated impaired tumor growth in vivo. CONCLUSION: Low dose NP promotes the growth of colon tumors through GPR30-mediated activation of ERK1/2 signaling.