lncRNA TUG1对卵巢癌细胞OVCAR3的增殖迁移及能量代谢的影响

Effects of lncRNA TUG1 on proliferation,migration and energy metabolism of OVCAR3 cells

目的探讨lncRNA TUG1在卵巢癌细胞中的表达及其对卵巢癌OVCAR3细胞增殖迁移及能量代谢的影响。方法采用实时荧光定量PCR(QPCR)法检测TUG1在卵巢上皮癌细胞(A2780、SKOV3、OVCAR3)及正常卵巢上皮细胞IOSE80中的表达。合成TUG1干扰片段转染OVCAR3细胞(干扰组),另设阴性对照组(NC组),QPCR验证TUG1的干扰效率,CCK-8法检测细胞增殖能力,Transwell和划痕实验检测细胞迁移能力,采用Seahorse能量分析仪检测细胞糖酵解压力及线粒体压力变化。结果 QPCR检测结果显示,与正常卵巢细胞IOSE80比较,卵巢癌SKOV3、OVCAR3细胞中的TUG1表达显著增加(P <0. 05)。CCK-8实验结果显示,NC组24、48 h时吸光值(A)值分别为0. 65±0. 01和0. 71±0. 01,显著高于干扰组的0. 48±0. 07和0. 50±0. 08,差异有统计学意义(P <0. 05)。干扰组48 h每个视野下穿膜细胞数为(46±15)个,显著少于NC组的(120±35)个;划痕实验也证实TUG1干扰后OVCAR3细胞迁移能力显著下降。Seahorse结果显示,干扰TUG1表达能显著降低OVCAR3细胞的线粒体呼吸和糖酵解通量。结论 lncRNA TUG1高表达可促进卵巢癌细胞的增殖迁移能力及能量代谢能力。

Objective To investigate the expression of lncRNA TUG1 in ovarian cancer cells and its effects on proliferation,migration and energy metabolism of ovarian cancer OVCAR3 cells.Methods The expression of TUG1 in ovarian epithelial cancer cells(A2780,SKOV3,OVCAR3)and normal ovarian epithelial cells(IOSE80)was detected by quantitative fluorescence polymerase chain reaction(QPCR).TUG1 interference fragment was synthesized and transfected into OVCAR3 cells(interference group),and the negative control group(NC group)was set up.QPCR was used to verify the interference efficiency of TUG1.CCK-8 assay was used to detect the cell proliferation,Transwell and scratch assay were used to detect the cell migration,and seahorse energy analyzer was used to detect the changes of cell glycolysis pressure and mitochondrial pressure.Results QPCR results showed that the expression of TUG1 in ovarian cancer SKOV3,OVCAR3 cells was significantly higher than that in normal ovarian cells IOSE80(P<0.05).CCK-8 test results showed that the absorbency(A)values of NC group at 24 and 48 hours were 0.65±0.01 and 0.71±0.01,which were significantly higher than those of interfering group at 0.48±0.07 and 0.50±0.08(P<0.05).The number of penetrating cells per field of vision in the interference group at 48 hours was 46±15,which was significantly less than 120±35 in the NC group.Scratch test also confirmed that OVCAR3 cell migration ability decreased significantly after TUG1 interference.Seahorse results showed that the decreased expression of TUG1 significantly reduced mitochondrial respiration and glycolysis flux in OVCAR3 cells.Conclusion High expression of lncRNA TUG1 can promote the proliferation,migration and energy metabolism of ovarian cancer cells.