microRNA-335对人非小细胞肺癌细胞迁移、侵袭及增殖能力的影响

Effect of microRNA-335 on migration,invasion and proliferation of NSCLC cell

目的:研究microRNA-335(miR-335)在非小细胞肺癌中的表达及其对细胞迁移、侵袭能力与增殖能力的影响。方法:采用荧光实时定量PCR检测miR-335在12对非小细胞肺癌与癌旁正常组织中的表达差异,以及miR-335在非小细胞肺癌SPCA-1细胞与肺正常上皮细胞16HBE中的表达差异;应用Lipofectamine 2000瞬时转染anti-miR-335下调SPCA-1细胞中miR-335的表达,并通过荧光实时定量PCR验证;划痕实验检测miR-335对SPCA-1细胞迁移能力的影响;Transwell侵袭实验检测miR-335对SPCA-1细胞侵袭能力的影响;MTT实验及克隆形成实验检测miR-335对SPCA-1细胞增殖能力的影响。结果:与癌旁正常组织比较,miR-335在非小细胞肺癌组织中显著高表达(P<0.05);与16HBE细胞比较,miR-335在SPCA-1细胞中显著高表达(P<0.05);利用Lipofectamine 2000瞬时转染anti-miR-335入SPCA-1细胞24 h时miR-335表达明显减弱(P<0.01);抑制miR-335表达对SPCA-1细胞迁移和侵袭能力有显著的抑制作用,抑制率分别为(42.8±2.7)%(P<0.01)及(73.25±4.4)%(P<0.01);抑制miR-335表达对SPCA-1细胞的增殖能力无明显影响。结论:miR-335在非小细胞肺癌中呈高表达,miR-335低表达能显著抑制SPCA-1细胞的迁移和侵袭能力,但对SPCA-1细胞的增殖能力无明显影响。

Objective：To explore the expression of microRNA-335 （miR-335） in non-small cell lung cancer（NSCLC） and its effect on migration,invasion and proliferation of NSCLC cell. Methods：Real-time quantitative PCR was used to detecte and compare the expression of microRNA-335 in 12 pairs of non-small cell lung cancer, adjacent normal tissue, NSCLC cell line SPCA-1 and normal epithelium cell line 16HBE. The expression of miR-335 in SPCA-1 cell was down-regulated by anti-miR-335 transient transfection with Lipofeetamine 2000, and the effect was identified by real-time quantitative PCR. Cell migration and invasion was measured in vitro by wound healing assay and transwell assay. The influence of miR-335 on SPCA-1 cell proliferation was detected by MTr rassay and colony formation assay. Results：Compared with adjacent normal tissues, miR-335 in non-small cell lung cancer was significantly up-regulated（P 〈 0.05）. Smilar to that,the expression of miR-335 was significantly increased in SPCA-1 cell compared to 16HBE cell （P 〈 0.05）. The miR-335 expression of SPCA-1 cell was significantly down-regulated 24 hours after anti-miR-335 transfection （P 〈 0.01）. In SPCA-1 cell,decreased miR-335 significantly inhibited cell migration and invasion,with the inhibiting rates of （42.8± 2.7）%（P 〈 0.01） and （73.25 ±4.4）% （P 〈 0.01）,respectively. But,no effect on SPCA-1 cell proliferation was observed when the expession of miR-335 was inhibited. Conclusion：The expression of miR-335 is up-regulated in NSCLC. Down-regulating expression of miR-335 significantly inhibites migration and invasion of SPCA-1 cell while has no effct on cell proliferation.