microRNA-1183通过抑的表达促进胃癌细胞的增殖及转移

来源 :中国医科大学学报 | 被引量 : 0次 | 上传用户:hasfyturnip
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目的 研究microRNA-1183对胃癌细胞增殖及转移的影响,初步探讨microRNA-1183和CBL-B信号转导通路在此过程中的作用.方法将人胃癌细胞MGC803瞬时转染microRNA模拟物microRNA-1183,采用实时PCR法检测胃癌细胞中microRNA-1183的表达,MTT法检测细胞增殖的能力,Transwell法检测细胞的转移.双荧光素酶报告基因检测系统检测microRNA-1183与CBL-B的结合,Western blotting观察蛋白的表达.结果MTT法显示microRNA-1183可促进胃癌细胞MGC803的增殖,Transwell法显示microRNA-118促进了胃癌细胞MGC803的转移.BLAST对比分析结果显示,CBL-B是microRNA-1183的靶基因之一.Western blotting结果显示,过表达microRNA-1183后,CBL-B的表达明显下调.双荧光素酶报告基因检测系统证实CBL-B是microRNA-1183的靶基因.敲除CBL-B后,促进了MGC803的增殖与转移.microRNA-1183通过抑制CBL-B促进了胃癌细胞MGC803的增殖与转移.结论microRNA-1183通过抑制CBL-B的表达,促进胃癌细胞MGC803的增殖及转移.“,”Objective To investigate the effect of microRNA-1183 on proliferation and metastasis on gastric cancer cells and to explore the role of microRNA-1183 and CBL-B signaling pathways in this process. Methods MGC803 cells were transfected with a microRNA-1183 mimic. Real-time PCR detected the expression of microRNA-1183 in gastric cancer cell line MGC803. MTT detected the proliferative effect of microRNA-1183 on MGC803 gastric cancer cells. A Transwell assay detected the effect of microRNA-1183 on the metastasis of MGC803 gastric cancer cells. A dual luciferase reporter assay detected the binding ability between microRNA-1183 and CBL-B. The expression of the protein was tested by Western blotting. Results MTT assay results showed that microRNA-1183 promoted the proliferation of MGC803 cells. Transwell assay results revealed that microRNA-1183 promoted the metastasis of MGC803 cells. The results of BLAST contrast analysis show that CBL-B is one of the target genes of microRNA-1183. Western blotting analysis showed that the mimic microRNA-1183 inhibited the expression of CBL-B. A dual luciferase reporter assay showed that CBL-B was the target gene of microRNA-1183. A CBL-B knockdown promoted the proliferation and metastasis of MGC803 cells. microRNA-1183 promoted the proliferation and metastasis of MGC803 cells by inhibiting the expression of CBL-B. Conclusion microRNA-1183 can inhibit the proliferation and metastasis of gastric cancer cell lines by inhibiting the expression of CBL-B.
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