沉默鞘磷脂合酶1小干扰RNA对卵巢癌HO8910细胞增殖、凋亡及核因子-κB通路的影响  

Effects of sphingomyelin synthase 1 small interfering RNA on proliferation, apoptosis and nuclear factor-kappa B pathway of ovarian cancer HO8910 cells

作　　者：任佳琪 林昌岫 杭月 张璇 胡月 历昕妤 黄成日 REN Jia-qi;LIN Chang-xiu;HANG Yue;ZHANG Xuan;HU Yue;LI Xin-yu;HUANG Cheng-ri(Department of Obstetrics and Gynecology,Yanbian University Hospital,Yanbian 133000,Jilin Province,China;Central Laboratory,Yanbian University Hospital,Yanbian 133000,Jilin Province,China)

机构地区：[1]延边大学附属医院妇产科,吉林延边133000 [2]延边大学附属医院中心实验室,吉林延边133000

出　　处：《中国临床药理学杂志》2020年第22期3683-3686,共4页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81360382);吉林省教育厅“十三五”科学技术课题资助项目(JJKH20191118KJ)。

摘　　要：目的探讨小干扰RNA(siRNA)沉默鞘磷脂合酶1(SMS1)表达对卵巢癌HO8910细胞增殖、凋亡及核因子-κB(NF-κB)通路的影响。方法将体外培养的人卵巢癌HO8910细胞随机分成空白组(不进行转染)、对照组(转染无义序列慢病毒载体)和实验组(转染SMS1 siRNA慢病毒载体)。3组细胞均在37℃、5%CO2饱和湿度条件下用含10%胎牛血清的DMEM培养基进行培养。转染48 h后,用实时荧光定量聚合酶链反应法检测细胞中SMS1 mRNA的表达情况,用噻唑蓝法检测细胞的增殖情况,用流式细胞术检测细胞的凋亡情况,用蛋白质印迹法检测细胞核和细胞质中NF-κB的表达情况。结果培养48 h后,空白组、对照组和实验组SMS1 mRNA表达水平分别为1.03±0.09,1.04±0.10和0.51±0.06,OD值分别为1.50±0.11,1.46±0.12和0.87±0.10,细胞凋亡率分别为(7.18±1.60)%,(7.07±1.54)%和(20.76±3.21)%,细胞核NF-κB蛋白表达水平分别为0.74±0.08,0.76±0.08和0.40±0.06,细胞质NF-κB蛋白表达水平分别为0.25±0.05,0.22±0.04和0.81±0.10,实验组的上述指标与空白组和对照组比较,差异均有统计学意义(均P<0.05)。结论沉默SMS1基因表达可能通过调节NF-κB通路,抑制卵巢癌细胞增殖,诱导其凋亡。Objective To investigate the influences of small interfering RNA(siRNA) silencing sphingomyelin synthase 1(SMS1) expression on proliferation, apoptosis, and nuclear factor-kappa B(NF-κB) pathway of ovarian cancer HO8910 cells. Methods Human ovarian cancer HO8910 cells cultured in vitro were randomly divided into three groups: blank group(no transfection), control group(transfected lentiviral vectors containing nonsense sequences), and experimental group(transfected lentiviral vectors containing SMS1 siRNA). The three groups were cultured in DMEM medium containing 10% fetal bovine serum at 37 ℃, 5%CO2 saturated humidity. After 48 hours of transfection, real-time quantitative polymerase chain reaction was used to detect the expression of SMS1 in cells, methyl thiazolyl tetrazolium assay was used to detect cell proliferation, flow cytometry was used to detect apoptosis, Western Blotting was used to detect the expressions of nucleus NF-κB and cytoplasm NF-κB proteins. Results After 48 hours of transfection,the expressions of SMS1 mRNA in blank group,control group and experimental group were1. 03 ± 0. 09,1. 04 ± 0. 10 and 0. 51 ± 0. 06,the OD values were 1. 50 ± 0. 11,1. 46 ± 0. 12 and 0. 87 ± 0. 10,the apoptosis rates were(7. 18 ± 1. 60) %,(7. 07 ± 1. 54) % and(20. 76 ± 3. 21) %,nucleus NF-κB protein was0. 74 ± 0. 08,0. 76 ± 0. 08 and 0. 40 ± 0. 06,cytoplasm NF-κB protein was 0. 25 ± 0. 05,0. 22 ± 0. 04 and0. 81 ± 0. 10,the differences between blank group and control group,experimental group were statistically significant(all P < 0. 05). Conclusion Silencing the expression of SMS1 gene may inhibit the proliferation and induce apoptosis of ovarian cancer cells by regulating the NF-κB pathway.

关 键 词：卵巢癌 鞘磷脂合酶1 细胞增殖 细胞凋亡 核因子-κB通路 

分 类 号：R97[医药卫生—药品]