miR-211通过抑制PTEN基因的表达促进肺鳞状细胞癌的增殖及侵袭能力

miR-211 proliferates lung squamous cell carcinoma proliferation and invasion through targeting PTEN gene

目的探讨miR-211是否通过调控PTEN的表达而影响人肺鳞状细胞癌细胞体外增殖和侵袭能力。方法通过双荧光素酶报告证明PTEN为miR-211的下游靶基因,miR-211及其阴性对照(miR-N. C.)转染人肺鳞状细胞癌细胞株NCI-H520,观察细胞中PTEN的mRNA水平和蛋白水平的表达变化;然后通过细胞增殖实验、细胞周期实验和侵袭实验(Trans-well),观察NCI-H520细胞体外增殖和侵袭能力的变化。结果与对照组相比,miR-211可显著降低荧光素酶的表达活性(70. 6%vs100%,χ2=28. 331,P=0. 001)。miR-211靶向作用PTEN可使NCI-H520细胞中mRNA表达水平较对照下调78. 9%(χ2=114. 973,P=0. 045),而蛋白水平则下调71. 4%(χ2=113. 318,P=0. 025)。miR-211处理后第3天,NCI-H520细胞的增殖能力比对照组细胞增高32. 6%(χ2=27. 143,P=0. 031)。而细胞周期实验表明与阴性对照相比,miR-7-5p处理组的NCI-H1975细胞,其S期所占比例显著上调(19. 2%vs32. 7%,χ2=5. 221,P=0. 013),而G0/G1期所占比例则显著降低(67. 8%vs55. 1%,χ2=2. 973,P=0. 026)。相比对照组,miR-211可将NCI-H520细胞的侵袭能力上调约47. 3%(χ2=48. 937,P=0. 035)。结论 MiR-211主要是通过靶向抑制PTEN表达增强人肺鳞状细胞癌的增殖和侵袭能力。

Objective To investigate whether the expression of PTEN regulated by miR-211 affects the pro liferation and invasion of human lung squamous cell carcinoma （LSCC）. Methods PTEN was validated as a downstream target gene of miR-211 by dual luciferase reporter assay. Human LSCC cell line NCI-H520 was transfected with miR-211 and negative control （miR-N. C. ） , then it used RT-RCR and western blot to detect the expression of PTEN in mRNA and protein level. In addition, the proliferation and invasion of NCI-H520 cells was observed through MTS cell proliferation assay, cell cycle assay and Trans-well invasion assay. Results Compared with the control group, miR-211 decreased significantly the luciferase activity of PTEN （70. 6% vs 100% , X2 = 28. 331, P = 0. 001）. The expression of PTEN was reduced by 78.9% and 71.4% in NCI-H520 cells line in mRNA and protein level, respectively. On the third dayafter miR-211 treatment, the proliferation of NCI-H1975 cells was increased about 32. 6% （ χ2 = 27. 143, P = 0. 003 ）. The cell cycle experiments showed that the percentage of S significantly increased （ 19.2% vs 32. 7% , χ2 = 5. 221, P = 0. 013） , and the proportion of GO/G1 phase decreased significantly （67. 8% vs 55.1% , χ2 = 2. 973, P = 0. 026） while compared with the negative control group. MiR-211 inhibited the invasion ability of NCI-H520 cells about 47. 3%. Conclusion miR-211 can inhibit the expression of PTEN resulting in proliferation of the proliferation and invasion of human LSCC.