LncRNA BANCR对卵巢癌细胞增殖、凋亡的影响及机制

Effect and Mechanism of LncRNA BANCR on Proliferation and Apoptosis of Ovarian Carcinoma Cells

目的研究长链非编码RNA BANCR(LncRNA BANCR)对卵巢癌细胞增殖及凋亡的影响,并探讨其机制。方法以实时荧光定量PCR(qRT-PCR)检测卵巢癌细胞系(A2780、A431、H08910、SKOV3)和正常卵巢上皮细胞系HOSE中LncRNA BANCR的表达。将卵巢癌SKOV3细胞系分为3组:沉默表达组(BANCR siRNA组)、阴性对照组(NT siRNA组)及空白对照组(Blank组),经Lipofectamine^TM 3000分别转染沉默序列pcDNA6.2-GW/BANCR siRNA、阴性对照序列pcDNA6.2-GW/NC scramble及空白质粒pcDNA6.2-GW。MTT法测定细胞活力,流式细胞术测定细胞凋亡,Western blot测定MAPK信号通路蛋白pERK、p-JNK、p-P38及凋亡相关蛋白Cleaved Caspase-3的表达。结果卵巢癌细胞系中LncRNA BANCR相对表达量均显著高于正常卵巢上皮细胞系HOSE。转染后,BANCR siRNA组的LncRNA BANCR表达量较NT-siRNA组和Blank组显著下调(均P<0.01)。转染后48、72及96 h,BANCR siRNA组细胞活力显著低于NT-siRNA及Blank组(P<0.05,P<0.01)。BANCR siRNA组细胞凋亡率显著高于NT-siRNA组及Blank组(均P<0.01)。LncRNA BANCR沉默后pERK、p-JNK蛋白表达下调,Cleaved Caspase-3蛋白表达上调,p-P38表达未受影响。结论沉默LncRNA BANCR表达可抑制卵巢癌细胞增殖、诱导凋亡,其机制可能与调控MAPK信号通路中的ERK和JNK表达有关。

Objective To study the effects of long-chain non-coding RNA BANCR(LncRNA BANCR)on proliferation and apoptosis of ovarian cancer cells,and to explore its mechanism.Methods Fluorescence real-time quantitative PCR(qPCR)was used to detect the expression of LncRNA BANCR in ovarian cancer cell lines(A2780,A431,H08910,SKOV3)and normal ovarian epithelial cell lines HOSE.SKOV3 cell lines were divided into three groups,silent expression group(BANCR siRNA group),negative control group(NT-siRNA group)and blank control group(Blank group).The silent sequence pcDNA6.2-GW/BANCR siRNA,negative control sequence pcDNA6.2-GW/NC scramble and blank plasmid pcDNA6.2-GW were transfected by Lipofectamine^ TM 3000,respectively.MTT assay was used to detect cell proliferation.Flow cytometry was used to detect cell apoptosis.Western blotting was used to detect the expression of pERK,p-JNK,p-P38 and Cleaved Caspase-3,which are the signal pathway proteins of MAPK.Results The relative expression of LncRNA BANCR in ovarian cancer cell lines was significantly higher than that in normal ovarian epithelial cell line HOSE.After transfection,the expression of LncRNA BANCR in BANCR siRNA group was significantly lower than that in NT-siRNA group and Blank group(P<0.01).At 48,72 and 96 h after transfection,A490 nm in BANCR siRNA group was significantly lower than that in NT siRNA and Blank group(P<0.05,P<0.01).The apoptotic rate of BANCR siRNA group was significantly higher than that of NT-siRNA group and Blank group(P<0.01).After LncRNA BANCR silencing,the expression of pERK and p-JNK protein was down-regulated,the expression of Cleaved Caspase-3 protein was up-regulated,and the expression of p-P38 was not affected.Conclusion Silencing the expression of LncRNA BANCR can inhibit the proliferation and induce apoptosis of ovarian cancer cells,which may be related to the regulation of ERK and JNK in the MAPK signaling pathway.