摘要
目的研究微小RNA(miR)-766-5p对人胃腺癌细胞(AGS)细胞增殖和放射敏感性的影响及潜在作用机制。方法以胃上皮细胞GES-1为对照,实时荧光定量逆转录聚合酶链式反应(qRT-PCR)检测胃癌细胞系AGS、MGC803、MKN-28和SGC-7901中组氨酸磷酸酶(LHPP)mRNA和miR-766-5p的表达,AGS细胞转染anti-miR-766-5p,确定抑制miR-766-5p表达对细胞的影响,克隆形成实验检测不同放射剂量处理后细胞存活分数,四甲基偶氮唑盐比色法(MTT)测定AGS细胞存活率,蛋白质印迹法(Westernblotting)检测细胞中LHPP、细胞周期蛋白D1(CyclinD1)和γ-H2AX蛋白表达,双荧光素酶报告系统验证miR-766-5p和LHPP的调控关系。结果与GES-1相比,miR-766-5p在胃癌细胞系AGS、MGC803、MKN-28和SGC-7901表达量升高[(3.30±0.24)、(2.89±0.19)、(2.05±0.21)、(1.99±0.19)比(1.00±0.10),P<0.05],LHPPmRNA[(0.85±0.09)、(1.01±0.11)、(0.75±0.08)、(0.99±0.10)比(4.46±0.25)]、蛋白表达量[(0.45±0.05)、(0.35±0.05)、(0.30±0.04)、(0.46±0.05)比(1.08±0.12)]降低(P<0.05);抑制miR-766-5p表达可抑制胃癌AGS细胞增殖并增强细胞的放射敏感性;miR-766-5p靶向负调控LHPP的表达;敲减LHPP可逆转anti-miR-766-5p对AGS细胞增殖和放射敏感性的作用。结论miR-766-5p通过靶向促进LHPP表达增强胃癌AGS细胞的放射敏感性并抑制癌细胞增殖。miR-766-5p有望成为胃癌临床放射增敏靶点。
Objective To investigate the effect of miR-766-5p on proliferation and radiosensitivity of gastric cancer AGS cells and its potential mechanism.Methods GES-1 cells were took as the control,the expression levels of LHPP mRNA and miR-766-5p in gastric cancer cell lines AGS,MGC803,MKN-28 and SGC-7901 were detected by qRT-PCR.AGS cells were transfected with anti-miR-766-5p to determine the effect of inhibiting the expression of miR-766-5p on AGS cells.The survival fraction of AGS cells treated with different doses of radiation was measured by colony formation assay.The cell viability was determined by MTT assay.The expression levels of LHPP,CyclinD1 andγ-H2AX protein were detected by Western blot.The relationship between miR-766-5p and LHPP was verified by the dual-luciferase reporter assay system.Results Compared with GES-1,the expression of miR-766-5p in gastric cancer cell lines AGS,MGC803,MKN-28 and SGC-7901 increased[(3.30±0.24),(2.89±0.19),(2.05±0.21),(1.99±0.19)vs.(1.00±0.10),P<0.05],and the expression of LHPP mRNA[(0.85±0.09),(1.01±0.11),(0.75±0.08),(0.99±0.10)vs.(4.46±0.25)]and protein[(0.45±0.05),(0.35±0.05),(0.30±0.04),(0.46±0.05)vs.(1.08±0.12)]decreased(P<0.05).Inhibition of miR-766-5p inhibited the proliferation and enhanced the radiosensitivity of AGS cells.MiR-766-5p negatively regulated the expression of LHPP.Knockdown of LHPP reversed the effects of inhibition of miR-766-5p on the proliferation and radiosensitivity of AGS cells.Conclusions MiR-766-5p enhances the radiosensitivity and promotes the proliferation of gastric cancer AGS cells by targeting LHPP.MiR-766-5p is expected to be a target for clinical radiosensitization of gastric cancer.
作者
刘国成
官一平
王成
黄国军
LIU Guocheng;GUAN Yiping;WANG Cheng;HUANG Guojun(Department of General Surgery,Pidu District People's Hospital,Chengdu,Sichuan 611730,China;Department of Gastroenterology,Pidu District People's Hospital,Chengdu,Sichuan 611730,China;Department of Medical Branch,Pidu District People's Hospital,Chengdu,Sichuan 611730,China;Department of Oncology,Pidu District People's Hospital,Chengdu,Sichuan 611730,China)
出处
《安徽医药》
CAS
2022年第2期274-278,共5页
Anhui Medical and Pharmaceutical Journal
