miR-9对人脑胶质瘤LN229细胞增殖的影响

Influence of miR-9 on the proliferation of LN229 glioma cells

目的探讨miR-9对人脑胶质瘤LN229细胞增殖的影响及可能机制。方法对数生长期人脑胶质瘤LN229细胞,随机分为miR-9抑制剂组(转染30 mg/L miR-9小干扰RNA质粒5μL)、空白对照组(转染空载质粒)、乳酸脱氢酶A(labeling recombinant lactate dehydrogenase A,LDHA)激动剂组(转染30 mg/L miR-9小干扰RNA质粒5μL及4 mg/L LDHA激动剂5μL)。转染后培养48 h,3组采用CCK-8法检测细胞增殖率;实时荧光定量PCR法检测miR-9 mRNA、LDHA mRNA相对表达量;氧化酶法测定葡萄糖消耗量;比色测定法测定乳糖产量。结果转染后培养48 h,miR-9抑制剂组、空白对照组、LDHA激动剂组细胞增殖率吸光度值(0.60±0.01、0.90±0.02、1.15±0.05)依次增高(P<0.05);miR-9 mRNA相对表达量在miR-9抑制剂组(0.52±0.01)、LDHA激动剂组(0.53±0.01)低于空白对照组(1.67±0.01)(P<0.05),miR-9抑制剂组与LDHA激动剂组比较差异无统计学意义(P>0.05);miR-9抑制剂组、空白对照组、LDHA激动剂组LDHA mRNA相对表达量(0.64±0.01、0.87±0.01、2.77±0.01)、葡萄糖消耗量[(0.02±0.01)、(0.36±0.01)、(0.77±0.01)nmol/L]、乳糖产量[(43.02±4.01)、(73.02±3.11)、(1562.00±5.44)nmol/L]均依次增高(P<0.05)。结论miR-9通过促进LDHA表达,增强肿瘤细胞有氧糖酵解,促进人脑胶质瘤LN229细胞增殖。

Objective To investigate the influence of miR-9 on the proliferation of LN229 glioma cells and its possible mechanism.Methods LN229 cells in logarithmic growth phase were randomly divided into miR-9 inhibitor group(transfected with 5μL of 30 mg/L miR-9 small interfering RNA plasmid),blank control group(transfected with empty vector plasmid),and labeling recombinant lactate dehydrogenase A(LDHA)agonist group(transfected with 5μL of 30 mg/L miR-9 small interfering RNA plasmid and 5μL of 4 mg/L LDHA agonist).After 48 h of culture,CCK-8 method was used to detect the proliferation rate,real-time fluorescence quantitative PCR technique was used to detect the relative expressions of miR-9 mRNA and LDHA mRNA,oxidase method was used to detect the glucose consumption,and colorimetric method was used to detect the lactose production.Results After 48 h of culture,the optical density values of cell proliferation rate increased gradually in turn in miR-9 inhibitor group,blank control group and LDHA agonist group(0.60±0.01,0.90±0.02,1.15±0.05)(P<0.05).The relative expression of miR-9 mRNA was lower in miR-9 inhibitor group(0.52±0.01)and LDHA agonist group(0.53±0.01)than that in blank control group(1.67±0.01)(P<0.05),and showed no significant difference between miR-9 inhibitor group and LDHA agonist group(P>0.05).The relative expressions of LDHA mRNA(0.64±0.01,0.87±0.01,2.77±0.01),glucose consumption((0.02±0.01),(0.36±0.01),(0.77±0.01)nmol/L)and lactose production((43.02±4.01),(73.02±3.11),(1562.00±5.44)nmol/L)increased gradually in turn in miR-9 inhibitor group,blank control group and LDHA agonist group(P<0.05).Conclusion miR-9 can promote the proliferation of LN229 glioma cells by promoting the expression of LDHA and aerobic glycolysis of tumor cells.