| Objective:The purpose of this study was to investigate the expression of DNMT3 A in renal carcinoma tissues on the postoperative pathologic characteristics and renal cell metastasis and recurrence.By overexpressing DNMT3 A gene,we investigated the relationship between DNMT3 A expression and everolimus resistance in 786-o,as well as the regulation mechanism of DNMT3 A expression on apoptosis related proteins Bcl-2 and growth related proteins ERK1/2.Methods:We had collected cancer tissues and adjacent tissues of 58 renal carcinoma patients from January 2015 to December 2017.Which were diagnosed as renal clear cell carcinoma after surgery.These patients were all from the first affiliated hospital of xia men university.To detect the expression of DNMT3 A gene in the cancer tissues and adjacent tissues,we measured the clinical renal clear cell carcinoma specimens by qRT-PCR(quantitative real-time PCR,qRT-PCR).In order to clear the relationship between the DNMT3 A expression level in the tumor and the clinicopathology of renal cell carcinoma,We performed statistical analysis combine clinicopathological results of the patients.To elucidate the mechanism of DNMT3 A promoting renal cell proliferation and the possible signaling mechanism of everolimus resistance at the molecular and cellular biological level,We transfected DNMT3 A overexpressed plasmids and treated 786-o cells with everolimus.Results:We detected the DNMT3 A expression in 58 renal clear cell carcinoma patients by qPCR.,the result showed that the DNMT3 A expression in cancer tissue were striking higher than tissue adjacent to carcinoma(P < 0.05),the expression of DNMT3 A levels were significant correlation with renal clear cell carcinoma pathological staging,pathological grading and tumor maximum diameter(P < 0.05),while there were no significant differences in gender,age,BMI(P > 0.05),these all indicates that the expression of DNMT3 A was associated with tumor pathology classification and renal carcinoma recurrence metastasis(P < 0.05).After overexpressing DNMT3 A in renal cell carcinoma cell line 786-0,the expression of p-ERK1/2 protein associated with tumor growth was increased,and the increase of p-ERK1/2 was more obvious after treatment with 5 ?M of everolimus(P< 0.05);CCK8 test showed that the proliferation level was significantly higher than that of the control group after transfection of the expression plasmid,and it was more obvious after the treatment with everolimus(P<0.01).The monoclonal formation experiment proved that after overexpressing DNMT3 A,The colony formation of DNMT3 A plasmid transfected group was significantly higher than that of the control group,and the difference was statistically significant(P<0.01)Conclusion:The DNMT3 A expression in renal cell carcinoma is obviously higher than adjacent tissues.The expression of DNMT3 A is closely related to the clinicopathological features of renal cell carcinoma.The renal clear cell carcinoma with high expression of DNMT3 A likely have a worse pathological grade.Overexpressing DNMT3 A gene can increase the proliferation of renal cancer cell line 786-o.Under the condition of everolimus treatment,renal cancer cells can mediate the feedback activation of downstream ERK1/2 through the expression of DNMT3 A,and promote the tolerance of renal cancer cells.The medicinal properties and the elucidation of related mechanisms provide new targets and therapeutic strategies for the treatment of renal cancer. |
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