Analsis on the protective effect and mechanism of Nec-1 on cytotoxicity induced by cyclosporine A / 中国生化药物杂志

Objective To investigate the protective effect and its potential molecular mechanism of Nec-1 on cytotoxicity induced by cyclosporine A.Methods MRTEpiC, glomerular endothelial cell MGEC and mesangial cell line MMC were co-administered with Nec-1 and cyclosporin A in mouse renal tubular epithelial cell line, and then MTT assay and soft agar clone formation assay were used to detect Cell growth curve changes, clonal formation ability.Apoptosis was detected by flow cytometry.The expression of cyclin D1, CDK4, CDK2, Cyclin E and apoptosis-related Caspase 3 were detected by Western blot.Results After cyclosporine A action, the cell growth ability was significantly decreased and the clone formation ability was significantly decreased(P<0.05).Cyclin D1, CDK4, CDK2 and Cyclin E were significantly increased(P<0.05), but the ratio of apoptosis and the expression of Caspase 3 did not change.Nec-1 has obvious protective effect on cytotoxicity induced by cyclosporine A, which can increase the cell growth ability and clone formation ability, and reduce the cell cycle-related proteins Cyclin D1, CDK4, CDK2, Cyclin E.Conclusion Nec-1 has cytotoxic effect on the glomeruli and renal tubular cells by up-regulating the cell cycle-related proteins Cyclin D1, CDK4, CDK2 and Cyclin E, while Nec-1 has protective effect.

Expression of Proteins Related Cell Cycle and Apoptosis during Diethylnitrosamine (DEN)-induced Hepatocarcinogenesis in Rats

PURPOSE: To explore the role of cell cycle and apoptosis regulators during hepatocarcinogenesis, the expression of cell cycle-related proteins (cyclin D1 and p27kip1) and apoptosis-related proteins (p53, survivin, caspase 3). METHODS: Sprague-Dawley rats were given 120 ppm diethylnitrosamine (DEN) as a carcinogen and sequentially sacrificed. The expression of cell cycle and apoptotic related proteins were examined by light microscopy and immunohistochemistry. RESULTS: During the DEN-induced hepatocarcinogenesis, sequential histologic changes from preneoplastic lesions (altered hepatic cellular foci, hyperplastic nodules, and hepatocellular adenomas) and ultimately overt hepatocellular carcinomas and metastatic lesions were noted. The cyclin D1 were progressively increased from preneoplastic lesions to hepatocellular carcinomas. However, the p27kip1 and the survivine proteins did not show any other difference with the increasing degree of carcinogenesis. The p53 and caspase 3 proteins were more significantly increased in hepatocellular carcinomas than preneoplastic lesions. The cyclin D1 protein expression did not show any correlation with the expression of p27Kip1 protein, but the p53 expression was related to the expression of survivin and caspase 3. CONCLUSION: From the above results, over-expression of cyclin D1 plays a role in the early and late stages of hepatocarcinogenesis. In addition p53 and caspase 3 might be useful markers for evaluating the risk of malignant transformation.

Significance of Expression of p16, Cyclin D1, Rb, and p53 Protein and Correlation with Clinicopathologic Prognostic Factors in Invasive Ductal Carcinoma of the Breast

The retinoblastoma (Rb)/cyclin D1/p16 pathway is an important constituent of cell cycle regulation. Perturbations in this pathway due to a variety of genetic aberrations have been reported in many human cancers including breast cancer. We examined the significance of immunoexpression of p16 protein, cyclin D1 protein, Rb protein (pRb), and p53 protein in 128 cases of invasive breast carcinoma. The results were correlated with survival rate and clinicopathological variables, including age, histologic grade, lymph node status, tumor size, estrogen receptor (ER), and progesterone receptor (PR) content. Abnormal expressions of p16 and pRb which were defined as negative staining were seen in 21% and 43% of tumors, respectively. There was a significant inverse relationship between p16 and pRb expression. There was no correlation between p16 staining and any other parameters, including survival rate, cyclin D1, p53, and clinicopathologic variables. Surprisingly, there was a trend for tumors which were positive for pRb to be grade III ductal carcinomas. Cyclin D1 positivity was noted in 46% of cases. The expression of cyclin D1 protein was significantly higher in lower histologic grade, higher ER and PR expression. The expression of p53 protein showed a significant correlation with high tumor grade. In a Cox multivariate analysis, neither p16, pRb, cyclin D1 nor p53 was an independent predictor, but tumor size and lymph node status were independent predictors of patient outcome.