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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 investigate the expression of microRNA-335 (miR-335) in non-small cell lung cancer and its effect on cell migration, invasion and proliferation. Methods: The expression of miR-335 in 12 non-small cell lung cancer tissues and adjacent normal tissues was detected by real-time fluorescence quantitative PCR and the expression of miR-335 in non-small cell lung cancer SPCA-1 cells and normal lung epithelial cells 16HBE The expression of miR-335 in SPCA-1 cells was transiently transfected with anti-miR-335 by Lipofectamine 2000. The expression of miR-335 was verified by real-time quantitative PCR. Scratch assay was used to detect the effect of miR-335 on the migration ability of SPCA- The effect of miR-335 on the invasion ability of SPCA-1 cells was detected by Transwell invasion assay. The effect of miR-335 on the proliferation of SPCA-1 cells was detected by MTT assay and clone formation assay. Results: miR-335 was highly expressed in non-small cell lung cancer (P <0.05) compared with adjacent normal tissues (P <0.05). Compared with 16HBE cells, miR-335 was significantly higher in SPCA-1 cells ; MiR-335 expression was significantly decreased (P <0.01) by transient transfection of anti-miR-335 into SPCA-1 cells with Lipofectamine 2000 for 24 h; inhibition of miR-335 expression significantly inhibited the migration and invasion ability of SPCA-1 cells (42.8 ± 2.7)% (P <0.01) and (73.25 ± 4.4)% respectively (P <0.01). Inhibition of miR-335 expression had no significant effect on the proliferation of SPCA-1 cells. CONCLUSIONS: miR-335 is highly expressed in non-small cell lung cancer. Low expression of miR-335 significantly inhibits the migration and invasion of SPCA-1 cells but has no effect on the proliferation of SPCA-1 cells.