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目的探讨微小RNA-769-5p(miR-769-5p)在非小细胞肺癌(NSCLC)细胞株中的表达及其对细胞增殖、迁移和侵袭能力的影响。方法采用实时荧光定量PCR(QPCR)技术检测miR-769-5p在5种NSCLC细胞(A549、NCI-H1299、NCI-H157、ANIP-973和GLC-82)中的相对表达量,选择相对表达量最低和最高的细胞株分别转染miR-769-5p mimics/miR-769-5p inhibitor,另设miRNA mimics control/inhibitor control对照组;采用MTS实验、克隆形成实验及Transwell小室实验检测miR-769-5p对NSCLC细胞增殖、克隆形成、侵袭和迁移能力的影响。结果 miR-769-5p在A549、NCI-H1299、NCI-H157、Anip-973及GLC-82细胞中的相对表达量分别为0.06、0.40、0.09、1.04和2.31。在miR-769-5p表达水平最低的A549细胞中瞬时转染miR-769-5p mimics,在miR-769-5p表达水平最高的GLC-82细胞中瞬时转染miR-769-5p inhibitor。MTS结果显示,与对照组比较,miR-769-5p mimics能够抑制A549细胞的增殖(P<0.05),而miR-769-5p inhibitor能够促进GLC-82细胞的增殖(P<0.05)。平板克隆实验结果显示,与对照组比较,miR-769-5p mimics能够抑制A549细胞的平板克隆形成数(124.7±14.7 vs.399.4±46.0,P<0.05);而miR-769-5p inhibitor能够促进GLC-82细胞的平板克隆形成数(555.7±29.7 vs.366.3±28.7,P<0.05)。Transwell实验结果显示,与对照组迁移和侵袭跨膜细胞数比较,miR-769-5p mimics能够抑制A549细胞的迁移和侵袭(94.4±18.1 vs.157.8±22.9,84.4±15.1 vs.135.6±16.7,P<0.05);miR-769-5p inhibitor能够促进GLC-82细胞的迁移和侵袭(226.7±40.3 vs.153.3±38.7,196.7±39.1 vs.138.9±40.4,P<0.05)。结论 miR-769-5p可以抑制NSCLC细胞的增殖、侵袭和迁移,可能成为NSCLC的潜在靶点。
Objective To investigate the expression of microRNA-769-5p (miR-769-5p) in non-small cell lung cancer (NSCLC) cell lines and its effect on cell proliferation, migration and invasion. Methods The relative expression of miR-769-5p in five NSCLC cell lines (A549, NCI-H1299, NCI-H157, ANIP-973 and GLC-82) was detected by real-time fluorescence quantitative PCR (QPCR) The lowest and the highest cell lines were transfected with miR-769-5p mimics / miR-769-5p inhibitor respectively, and another miRNA mimics control / inhibitor control group was established. MTS assay, clone formation assay and Transwell chamber assay were used to detect miR- 5p on the proliferation, colony formation, invasion and migration of NSCLC cells. Results The relative expression levels of miR-769-5p in A549, NCI-H1299, NCI-H157, Anip-973 and GLC-82 cells were 0.06, 0.40, 0.09, 1.04 and 2.31, respectively. The miR-769-5p mimics were transiently transfected into A549 cells with the lowest expression level of miR-769-5p and miR-769-5p transiently transfected into GLC-82 cells with the highest expression level of miR-769-5p. The results of MTS showed that miR-769-5p mimics could inhibit the proliferation of A549 cells (P <0.05) compared with the control group, while miR-769-5p inhibitor could promote the proliferation of GLC-82 cells (P <0.05). The result of plate clone assay showed that miR-769-5p mimics could inhibit the number of A549 cells (124.7 ± 14.7 vs.399.4 ± 46.0, P <0.05) compared with the control group, while miR-769-5p inhibitor promoted Platelet clone formation numbers of GLC-82 cells (555.7 ± 29.7 vs. 366.3 ± 28.7, P <0.05). The results of Transwell assay showed that miR-769-5p mimics inhibited the migration and invasion of A549 cells compared with the control group (94.4 ± 18.1 vs.157.8 ± 22.9, 84.4 ± 15.1 vs.135.6 ± 16.7, P <0.05). MiR-769-5p inhibitor promoted the migration and invasion of GLC-82 cells (226.7 ± 40.3 vs 153.3 ± 38.7, 196.7 ± 39.1 vs.138.9 ± 40.4, P <0.05). Conclusion miR-769-5p can inhibit the proliferation, invasion and migration of NSCLC cells and may be a potential target for NSCLC.