肺癌细胞中miR-182的功能及作用机制

Functions of miR-182 in Lung Cancer Cell and Its Mechanism of Action

目的检测miR-182在肺癌细胞中的表达和对肺癌细胞生物学特性的影响,并观察miR-182对靶基因profilin1(PFN1)表达的影响。方法 qRT-PCR检测miR-182在肺癌细胞中的表达。MTT和流式细胞仪分别检测miR-182对95-D细胞活力、增殖和凋亡的影响。Transwell侵袭小室检测miR-182对95-D细胞侵袭的影响。荧光素酶报告基因系统验证PFN1基因。Western blot方法检测miR-182对细胞中PFN1蛋白表达的影响。结果 qRT-PCR结果显示:miR-182在肺癌细胞中的表达显著上升(P<0.01)。MTT和流式细胞仪分析结果显示:抑制miR-182表达后,细胞活力和增殖指数明显下降(P<0.05);细胞的凋亡显著增加(P<0.01)。Transwell侵袭小室检测结果显示:抑制miR-182表达后,细胞侵袭能力显著降低(P<0.05)。Western blot结果显示:抑制miR-182表达后,靶蛋白PFN1的表达显著上升(P<0.05)。结论 miR-182在肺癌细胞中表达上调,并促进95-D细胞增殖和侵袭,并可负性调节PFN1蛋白的表达。

Objective To determine the expression of miR-182 in human lung cancer cell lines, and to evaluate the effects of miR-182 on biological characteristics and targeting profilin1(PFN1) of the lung cancer cell line.Methods The expression of miR-182 was detected by qRT-PCR in lung cancer cells. The infl uence of miR-182 on the viability, proliferation and apoptosis of 95-D cells was evaluated by MTT and fl ow cytometry assay. The role of miR-182 in the invasive potential of 95-D cells was studied by Transwell chamber assay. A luciferase reporter gene system was used to verify that PFN1 is a target gene for miR-182. The resulting effects of miR-182 on PFN1 protein expression was verified by Western blot analysis. Results By qRT-PCR, the expression of miR-182 was higher in lung cancer cells than that in HBE cells(P<0.01). Following a depression of miR-182 expression, cell viability, proliferation index and invasive potential were decreased(P<0.05), and apoptotic index and PFN1 protein levels were increased(P<0.01); the invasion was signifi cantly decreased(P<0.05).Conclusion The expression of miR-182 is upregulated in lung cancer cells. miR-182 negatively regulates PFN1 protein expression and promotes the proliferation and invasion of lung cancer cells.