沉默XIST通过靶向miR-185抑制人胶质瘤U87细胞增殖

Knockdown of XIST inhibits proliferation of human U87 glioma cell by targeting miR-185

目的探讨沉默长链非编码RNA XIST对人胶质瘤U87细胞增殖的影响及可能机制。方法 Lipo3000将XIST沉默质粒载体转染胶质瘤U87细胞后,Real-Time PCR检验转染效率;CCK-8法检测沉默XIST对U87细胞增殖的影响;Real-Time PCR检测转染XIST沉默载体后,U87细胞中miR-185的表达水平变化;双荧光素酶报告基因实验验证XIST与miR-185存在靶向结合;Lipo3000将miR-185过表达和沉默质粒载体分别转染U87细胞,Real-Time PCR检验转染效率;CCK-8法检测转转染miR-185后,U87细胞增殖能力的变化;Lipo3000将miR-185过表达和沉默质粒载体分别转染建立完成的XIST沉默U87细胞系,应用CCK-8法检测U87细胞的增殖水平变化。结果 XIST沉默显著降低了U87细胞的增殖能力,显著提高了miR-185在U87细胞中的表达水平。验证了XIST和miR-185存在着靶向结合。miR-185沉默显著提高了U87细胞增殖能力;显著阻断了沉默XIST下调U87细胞增殖能力的作用;miR-185过表达则作用相反。结论沉默XIST能够通过靶向miR-185,抑制胶质瘤U87细胞增殖。

Objective To study the effect and potential mechanism of glioma U87 cells proliferation regulated by knocking down long non-coding RNA XIST. Methods Lipo3000 was used to transfect Glioma U87 cells with XIST silencing plasmid vector, the efficiency of transfection had been tested by real-time PCR, CCK-8 was used to evalutate proliferation of U87 glioma ; Real-Time PCR assay was used to assess the expression of miR-185 in U87 glioma changed with knocking down XIST. Dual-Luciferase Reporte Assay was used to verify the combination of XIST and miR- 185. miR- 185 overexpression and and silencing plasmid vectors were transfected to U87 glioma cell respectiverly, the efficience of transfection had been tested by Real-time PCR, CCK-8 was used to detect proliferation of U87 glioma cell. miR-185 overexpression and and silencing plasmid vectors were transfected to XIST silencing U87 glioma cell line, CCK-8 was used to detect the variation of proliferation in U87 glioma. Results Silencing XIST significantly inhibited the proliferation of U87 cells, obviously improved the expression level of miR-185 in U87 glioma; verified the combination of XIST and miR-185; silencing miR-185 markedly improved the proliferation of glioma U87 ceils and obviously blocked the effect of inhibiting proliferation of U87 ceils by silencing XIST. Conclusion Knockdown of XIST inhibits proliferation of human U87 glioma cell by targeting miR-185.