lncRNA MAFG-AS1靶向miR-3180-3p调控口腔鳞癌HSC4细胞增殖和凋亡

lncRNA MAFG-AS1 regulates proliferation and apoptosis of oral squamous cell carcinoma HSC4 cells by targeting miR-3180-3p

目的:探讨长链非编码RNA(lncRNA)MAFG反义RNA1(MAFG-AS1)靶向miR-3180-3p对口腔鳞癌细胞增殖和凋亡的影响。方法:实时定量PCR检测口腔鳞癌组织和对应正常黏膜中MAFG-AS1和miR-3180-3p表达。以口腔鳞癌细胞HSC4为研究对象,分别构建干扰MAFG-AS1、过表达miR-3180-3p或抑制miR-3180-3p表达的口腔鳞癌细胞株,CCK-8、平板克隆实验、流式细胞术、蛋白印迹法检测细胞活力、克隆形成数、凋亡率及cleaved-caspase 3、cleaved-caspase 9表达。荧光素酶报告实验验证MAFG-AS1和miR-3180-3p的靶向关系。结果:与正常黏膜比较,口腔鳞癌组织MAFG-AS1表达显著升高(P<0.05),miR-3180-3p表达显著降低(P<0.05)。干扰MAFG-AS1或过表达miR-3180-3p均可降低HSC4细胞活力和克隆形成数(P<0.05),促进细胞凋亡(P<0.05),上调cleaved-caspase 3和cleaved-caspase 9蛋白表达(P<0.05)。miR-3180-3p是MAFG-AS1的直接靶点,MAFG-AS1负调控miR-3180-3p表达。抑制miR-3180-3p可增加HSC4细胞活力和克隆形成数(P<0.05),抑制细胞凋亡(P<0.05),下调cleaved-caspase 3和cleaved-caspase 9蛋白表达(P<0.05),进而削弱干扰MAFG-AS1对HSC4细胞的增殖抑制和凋亡促进作用(P<0.05)。结论:干扰MAFG-AS1通过上调miR-3180-3p表达抑制口腔鳞癌HSC4细胞增殖,促进细胞凋亡。

Objective:To investigate effect of long non-coding RNA(lncRNA)MAFG antisense RNA1(MAFG-AS1)targeting miR-3180-3p on proliferation and apoptosis of oral squamous cell carcinoma.Methods:MAFG-AS1 and miR-3180-3p expressions in oral squamous cell carcinoma tissues and corresponding normal mucosa were detected by real-time quantitative PCR.Oral squamous cell carcinoma HSC4 was used as research object to construct oral squamous cell carcinoma lines that interfered with MAFG-AS1,overexpressed miR-3180-3p or miR-3180-3p inhibition.Cell viability,clonal formation numbers,apoptosis rate,cleaved cleaved-caspase 3 and cleaved-caspase 9 expressions were detected by CCK-8,plane cloning experiment,flow cytometry and Western blot,respectively.Targeting relationship between MAFG-AS1 and miR-3180-3p was assessed with luciferase reporter experiment.Results:Compared with normal mucosa,MAFG-AS1 expression was remarkably increased(P<0.05),while miR-3180-3p expression was remarkably decreased(P<0.05)in oral squamous cell carcinoma tissues.Interference with MAFG-AS1 or miR-3180-3p overexpression could reduce HSC4 cell viability and clonal formation numbers(P<0.05),promote cell apoptosis(P<0.05),and up-regulate expressions of cleaved-caspase 3 and cleaved-caspase 9 proteins(P<0.05).miR-3180-3p was direct target of MAFG-AS1,and MAFG-AS1 negatively regulates miR-3180-3p expression.miR-3180-3p inhibition could increase HSC4 cell viability and colony formation number(P<0.05),inhibit cell apoptosis(P<0.05),and down-regulate cleaved-caspase 3 and cleaved-caspase 9 protein expressions(P<0.05),thereby weakening effect of interference with MAFG-AS1 on proliferation inhibition and apoptosis promotion on HSC4 cells(P<0.05).Conclusion:Interfering with MAFG-AS1 can inhibit proliferation and promote apoptosis of oral squamous cell carcinoma cells HSC4 by up-regulating miR-3180-3p expression.