Study On TET1 Expression In Renal Carcinoma And And Its Regulation On Cancer Cell Behavior

Renal cell carcinoma (RCC) is one of the most common malignant tumors in urinary system. As it has very complicated biological behavior, the mechanism of tumorigenesis and metastasis is still unknown. It has been reported that gene methylation was related to invasion and metastasis. TET1 was documented as a demethylation factor that down-regulated in several tumors, including hepatocellular carcinoma, squamous cell lung carcinoma, prostate cancer and bladder cancer. However, there is limited study on the functions and mechanisms of TET1 in RCC. Therefore, the study involving the investigation of TET1 effect on RCC was an insight to discover effective prognostic biomarker and therapeutic target of RCC.Part 1 Expressions of TET1 in RCC specimanObjective:To determined TET1 levels in renal tumor tissues, matched adjacent non-tumor tissues..Methods:TET1 levels were measured by qRT-PCR, Western bloting and immunohistochemistry in 82 pairs of tumor tissues and matched adjacent non-tumor ones.Results:The expression level of TET1 mRNA was remarkably down-regulated in cancer tissue compared with that in adjacent cancer tissue (p<0.05). The expression level of TET1 protein was significantly lower in cancer tissue than that in adjacent cancer tissue by immuohistochemistry and Western blotting (p<0.05).Conclusion:TET1 is remarkably down-regulated in RCC tumor tissues.Part 2 Clinical significance of TET1 in RCCObjective:To investigate whether TET1 levels correlated with the prognosis of RCC patients.Results:Kaplan-Meier analysis revealed that lower expression of TET1 level was correlated with noorer survival of RCC natientsConclusions:Lower expression of TET1 level in RCC tumor tissue is correlated with poorer survival of RCC patients.Part 3 TET1 expression in RCC cell line and relationship with cell behaviorObjective:To investigate TET1 expression in RCC cell line and its relationship with cell behavior..Methods:The expressions of TET1 protein in RCC cell line ACHN and HEK 293 T were assessed by Western Blotting. CCK-8 and Transwell assay were performed to evaluate cell proliferation and invasion.Results:Compared with 293T cells, the expression of TET1 in ACHN cells was significantly down-regulated. The capability of proliferation and invasion of ACHN cell line were significantly higher than that of 293T cells.Conclusion:RCC cell line had low level of TET1 expression, increased proliferation and invasion than 293 T cell, suggesting that TET1 might be implicated in the transformation and progress of RCC.Part 4 The effect of TET1 over-expression on apoptosis and invasion of RCCsObjective:Analysing the effect of TET1 on apoptosis and proliferation of RCC cells.Methods:RCC cell line ACHN was transfected with TET1 over-expression plasmid and vector, then Western Blotting was performed to validate the efficacy of overexpression. Annexin V and PI stain were conducted to compare the apoptosis of ACHN cells by flow mentry. Transwell assay was used for measurement the invasion capacility of ACHN cells.Results:TET1 over-expression inhibited the RCC cell proliferation and promoted cell apoptosis.Conclusion:TET1 regulates the RCC cell proliferation and apoptosis.