【摘要】 目的:探讨长链非编码RNA(lncRNA)人浆细胞瘤转化迁移基因1(PVT1)调控表皮生长因子受体(EGFR)/有丝分裂原活性蛋白激酶(MAPK)通路对卵巢癌细胞的影响。方法:将人卵巢癌细胞SKOV-3分为对照组(空白未处理)、si-NC组(转染携带空载体慢病毒)、si-lncRNA PVT1组(转染lncRNA PVT1 siRNA)。另选取人正常卵巢上皮细胞系HOSE为正常组。采用RT-qPCR检测细胞中lncRNA PVT1表达。采用Transwell实验、平板克隆实验分别检测SKOV-3细胞侵袭以及克隆形成能力。采用流式细胞术检测SKOV-3细胞凋亡情况。采用Western blot检测细胞中EGFR、细胞外信号调节蛋白激酶1/2(ERK1/2)、p-ERK1/2蛋白表达。根据细胞分组，将BALB/c裸鼠也相应随机分为对照组、si-NC组、si-lncRNA PVT1组，每组8只。建立裸鼠皮下移植瘤模型。比较各组裸鼠不同时间皮下移植瘤体积及平均瘤体重量。结果:对照组SKOV-3细胞lncRNA PVT1表达水平高于si-lncRNA PVT1组，si-lncRNA PVT1组SKOV-3细胞lncRNA PVT1表达水平高于正常组人卵巢正常上皮细胞系HOSE(均P<0.05)。与对照组及si-NC组比较，si-lncRNA PVT1组SKOV-3细胞克隆形成率、侵袭细胞数、EGFR、p-ERK1/2蛋白表达水平降低，细胞凋亡率升高(均P<0.05)。与对照组及si-NC组比较，si-lncRNA PVT1组裸鼠28 d时皮下移植瘤体积以及平均瘤体重量降低(均P<0.05)。结论:lncRNA PVT1在卵巢癌细胞中高表达，干扰其表达可抑制卵巢癌细胞侵袭、克隆、凋亡以及裸鼠皮下成瘤速度，作用机制可能与调控EGFR/MAPK信号通路有关。更多还原

【Abstract】 Objective:To explore the effect of long non-coding RNA(lncRNA) PVT1 on ovarian cancer cells by regulating epidermal growth factor receptor(EGFR)/mitogen-activated protein kinase(MAPK) pathway.Methods:Human ovarian cancer cells SKOV-3 were divided into control group(blank),si-NC group(transfected with empty carrier lentivirus) and si-lncRNA PVT1 group(transfected with lncRNA PVT1 siRNA).The normal human ovarian epithelial cell line HOSE was selected as the normal group.The expression of lncRNA PVT1 in cells was detected by RT-qPCR.Transwell assay and plate cloning assay were used to detect the invasion and cloning ability of SKOV-3 cells.The apoptosis of SKOV-3 cells was detected by flow cytometry.The expression of EGFR,extracellular signal-regulated protein kinase 1/2(ERK1/2) and p-ERK1/2 proteins were detected by Western blot.According to cell grouping, BALB/c nude mice were randomly divided into control group, si-NC group and si-lncRNA PVT1 group, with 8 mice in each group.The model of subcutaneous tumor transplantation in nude mice was established.The tumor volume and average tumor weight of subcutaneous transplantation were compared in each group.Results:lncRNA PVT1 expression level in SKOV-3 cells of control group was higher than that in si-lncRNA PVT1 group, and lncRNA PVT1 expression level in SKOV-3 cells of si-lncRNA PVT1 group was higher than that in HOSE of human ovarian normal epithelial cell line of normal group(all P<0.05).Compared with the control group and the si-NC group, the clonal formation rate of SKOV-3 cells, the number of invasive cells, the expression levels of EGFR and p-ERK1/2 protein in si-lncRNA PVT1 group were decreased, and the apoptosis rate was increased(all P<0.05).Compared with the control group and the si-NC group, the subcutaneous graft tumor volume and average tumor weight of nude mice in the si-lncRNA PVT1 group at 28 days were decreased(all P<0.05).Conclusion:LncRNA PVT1 is highly expressed in ovarian cancer cells, and interfering with its expression can inhibit the invasion, colony formation, apoptosis of ovarian cancer cells, and the speed of subcutaneous tumor formation in nude mice, which may be related to the regulation of the EGFR/MAPK signaling pathway.更多还原