长链非编码RNASNHG5在卵巢癌组织中表达及对卵巢癌细胞自噬影响  

Expression of long non-coding RNA SNHG5 in ovarian cancer tissues and its effect on autophagy of ovarian cancer cells

作　　者：沈晓萍[1] 张金春[1] SHEN Xiaoping;ZHANG Jinchun(Chongming Branch, Xinhua Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, 202150)

机构地区：[1]上海交通大学医学院附属新华医院崇明分院,202150

出　　处：《中国计划生育学杂志》2020年第11期1731-1735,1935,共6页Chinese Journal of Family Planning

摘　　要：目的:观察卵巢癌组织中长链非编码RNA(lncRNA)核仁小RNA宿主基因5(SNHG5)的表达,并分析SNHG5表达对卵巢癌细胞自噬的影响。方法:选取2018年8月—2019年3月本院卵巢癌手术取得的卵巢癌组织73例(卵巢癌组),卵巢囊肿手术取得卵巢组织82例(囊肿组),实时荧光定量PCR法(qRT-PCR)检测两组组织中SNHG5表达并分析与卵巢癌患者临床病理关系。培养人卵巢癌细胞SKOV3作为对照组,应用Lipofectamine 2000试剂将转染空质粒pcDNA3.1(+)、pcDNA3.1-SNHG5重组质粒的SKOV3细胞分别作为阴性对照组、SNHG5过表达组。细胞计数试剂盒(CCK8)法检测各组SKOV3细胞增殖情况,通过透视电镜、单丹(磺)酰戊二胺(MDC)法检测各组SKOV3细胞自噬水平,应用免疫印迹法(WB)检测LC3-I、LC3-II、Beclin 1蛋白表达水平。结果:lnc RNA-SNHG5相对表达量卵巢癌组(0.49±0.06)低于囊肿组(1.13±0.23),且卵巢癌组组织学分级G3、FIGO分期III^IV期中lnc RNA-SNHG5低表达者占比更高(P<0.05);SNHG5相对表达量SNHG5过表达组(1.97±0.37)高于空白组(1.03±0.20)和阴性对照组(1.05±0.21)(P<0.05);转染48h、72h、96h时SKOV3细胞增殖抑制率SNHG5过表达组(23.64±4.73%、43.57±8.71%、57.82±11.56%)高于空白组和阴性对照组(P<0.05);与空白组和阴性对照组比较,MDC阳性细胞占比SNHG5过表达组(42.38±7.64)最高,LC3-I蛋白表达量(0.16±0.03)降低,LC3-II(0.86±0.18)、beclin 1(0.97±0.18)升高(均P<0.05)。结论:lncRNA-SNHG5在卵巢癌组织中低表达,lncRNA-SNHG5过表达可明显抑制人卵巢癌细胞活力,促进细胞自噬。Objective:To observe the expression of long non-coding RNA(lncRNA)nucleolar small RNA host gene 5(SNHG5)in ovarian cancer tissues,and to analyze the effect of SNHG5 expression on autophagy of ovarian cancer cells.Methods:73 women with ovarian cancer were selected in group A and 82 women with ovarian cyst were selected in group B from August 2018 to March 2019.The expression of SNHG5 in ovarian tissues of women in the two groups was detected by real-time fluorescence quantitative PCR(qRT-PCR),and the relationships between the expression of SNHG5 in ovarian cancer tissue and clinical pathology of women in group A was analyzed.SKOV3 cells were cultured as blank control group,and SKOV3 cells transfected with empty plasmid pcDNA3.1(+)and recombinant plasmid pcDNA3.1-SNHG5 by Lipofectamine 2000 reagent were in negative control group and SNHG5 over expression group respectively.Cell counting kit(CCK8)method was used to detect the proliferation of SKOV3 cells in each group,the autophagy level of SKOV3 cells in each group was detected by fluoroscopy and electron microscopy,and the expression levels of LC3-I,LC3-II and Beclin 1 protein were detected by immunoblotting(WB).Results:Compared with that(1.13±0.23)in group B,the relative expression(0.49±0.06)of LNC RNA-SNHG5 in group A was significant lower(P<0.05).Compared with that in histological grade G1-G2 and FIGO stage I-II,the percentage of patients with high expression of LNC RNA-SNHG5 in histological grade G3 and FIGO stage III-IV was significant higher(P<0.05).Compared with that(1.03±0.20)in blank control group and that(1.05±0.21)in negative control group,the relative expression of SNHG5 in SNHG5 over expression group(1.97±0.37)had increased significantly(P<0.05).The inhibition rates of SKOV3 cell proliferation in SNHG5 over expression group at 48h,72h and 96h after transfection were 23.64±4.73%,43.57±8.71%,and 57.82±11.56%,respectively,which were significant higher than those in blank and negative control group(P<0.05).Compared with those in the blank group or

关 键 词：卵巢癌 长链非编码RNA 核仁小RNA宿主基因5 自噬 微管相关蛋白轻链3-β 

分 类 号：R73[医药卫生—肿瘤]