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论著:胃癌组织中microRNA 650的表达及其生物学作用和临床意义
Role of MicroRNA 650 in Gastric Cancer and Its Clinical Significance
霍守俊 薛锋 孔凡志 李芳 单远州 周联明 张学利
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作者单位:苏州大学
中文关键字:胃癌;凋亡;增殖;MicroRNA 650;G 蛋白信号调节因子7
英文关键字:Gastric cancer; Apoptosis; Proliferation; MicroRNA 650; Regulator of G protein signaling 7
中文摘要:目的:研究microRNA 650(miR 650)在胃癌组织中的表达及其与胃癌细胞增殖、凋亡及转移的关系。方法:采用逆转录聚合酶链式反应(reverse transcription polymerase chain reaction, RT PCR)检测miR 650在胃癌组织标本及胃癌细胞株中的表达量,胃癌细胞株转染miR 650模拟剂以及miR 650抑制剂 LNA寡核苷酸后采用CCK 8试剂盒检测细胞增殖情况,采用流式细胞仪检测细胞凋亡情况,采用LC MS/MS系统定量分析筛选miR 650靶蛋白,采用蛋白质印迹检测miR 650靶蛋白RGS7的表达水平。采用裸鼠异种移植模型验证miR 650对胃癌细胞的生物学作用及其抑制剂的生物疗效。结果:胃癌细胞株KATO III、NUGC4及SGC 7901高表达miR 650,而NCI N87及MKN 45仅微量表达miR 650。在转染miR 650抑制剂 LNA寡核苷酸后胃癌细胞增殖水平显著下降,凋亡水平显著上升。G 蛋白信号调节因子7(regulator of G protein signaling 7, RGS7)是miR 650下游靶点之一,靶蛋白RGS7表达水平与miR 650表达水平呈负相关, miR 650抑制剂可抑制肿瘤生长。结论:胃癌细胞株中miR 650的高表达可促进胃癌细胞增殖及转移并抑制其凋亡,机制可能是通过抑制靶蛋白RGS7的表达。
英文摘要:Objective: To explore the role of microRNA 650(miR 650) in the development and prognosis of gastric cancer and its clinical significance. Methods:Real time quantitative reverse transcription polymerase chain reaction was used to detect miR 650 expression levels in five gastric cancer cell lines and gastric cancer tissues. After the miR 650 mimic and inhibitor were transfected into gastric cancer cells, the cellular growth activity was estimated by CCK 8 kit; the apoptosis activity was tested by flow cytometry; the candidate targets of miR 650 were analysed using the LC MS/MS based techniques; western blot was used to test the expression levels of reglator of G protein signaling 7(RGS7) which acted as a target of miR 650 in gastric cancer cells. Nude mouse xenograft model was used to explore the biological effect of miR 650 in gastric cancer cells and the curative effect of miR 650 inhibitor. Results: High levels of miR 650 have been detected in KATO III, NUGC4 and SGC 7901 gastric cancer cell lines, while NCI N87 and MKN 45 gastric cancer cell lines only displayed low expression levels of miR 650. After transfection of LNA inhibitor, gastric cancer cell lines proliferation was significantly inhibited, while apoptosis was promoted. RGS7 was one of the functional downstream targets of miR 650, and the expression level of RGS7 was inversely correlated with the expression level of miR 650. MiR 650 inhibitor can suppress tumor growth. Conclusions: The up regulation of miR 650 promotes proliferation, metastasis and inhibits apoptosis of gastric cancer cells, and possibly through suppressing the expression of RGS7.
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