摘要
目的:研究TIRAP在上皮性卵巢癌细胞中的表达情况,及其对上皮性卵巢癌细胞增殖和凋亡能力的影响。方法:通过实时荧光定量PCR(qRT-PCR)和Western blot检测各组细胞中TIRAP的表达。利用CCK-8法、流式细胞技术探究TIRAP的表达对人上皮性卵巢癌细胞增殖和凋亡的影响。利用Western blot、ELISA法探究TIRAP的表达对p-NF-κB p65及下游免疫分子IL-6、TNF-α水平的影响。结果:与正常卵巢上皮细胞(IOSE-80)相比,上皮性卵巢癌细胞(SKOV-3,OVCAR3,A2780)中TIRAP的mRNA和蛋白表达显著升高。转染TIRAP siRNA后,SKOV-3细胞中TIRAP的表达显著降低;CCK-8实验显示:转染培养第96 h时,si-TIRAP组OD值(1.85±0.06)较Blank组(2.42±0.16)及Scramble control组(2.50±0.10)明显降低;细胞凋亡检测显示:si-TIRAP组凋亡率明显高于对照组;Western blot实验显示:LPS+si-TIRAP组p-NF-κB p65的表达与对照组相比显著降低;ELISA实验显示:LPS+si-TIRAP组IL-6、TNF-α的含量与对照组相比显著下降。结论:TIRAP在上皮性卵巢癌细胞中的表达升高,可能是上皮性卵巢癌细胞增殖的机制之一;下调TIRAP的表达可以抑制上皮性卵巢癌细胞增殖,促进细胞凋亡,并且通过抑制NF-κB通路的活化降低其下游免疫分子的表达。
Objective:To investigate the express of TIRAP in human epithelial ovarian cancer cell and the effect of TIRAP on proliferation and apoptosis of epithelial ovarian cancer cell.Methods:The expression level of TIRAP in cell from each treatment group was detected by Real-time PCR(qRT-PCR)and Western blot.CCK-8 method and flow cytometry were used to investigate the effect of TIRAP expression on proliferation and apoptosis of human epithelial ovarian cancer cells.Western blot and ELISA were used to investigate the effect of TIRAP expression on p-NF-κB p65 and the levels of downstream immune molecules IL-6 and TNF-α.Results:Compared with normal ovarian epithelial cells(IOSE-80),the expression of TIRAP in epithelial ovarian cancer cells(SKOV-3,OVCAR3,A2780)were significantly increased.After transfection with TIRAP siRNA,the expression of TIRAP in SKOV-3 cells was significantly decreased.CCK-8 experiment showed that the cell proliferation ability of TIRAP siRNA group was significantly reduced after transfection for 96 h and the OD value of the si-TIRAP group(1.85±0.06)was lower than that in the blank group(2.42±0.16)and the scramble control group(2.50±0.10).Apoptosis detection showed that the apoptosis rate in si-TIRAP group was significantly higher than that in control group.Western blot analysis showed that the expression of p-NF-κB p65 in LPS+si-TIRAP group was significantly lower than that in the control group.ELISA showed that the levels of IL-6 and TNF-αin the LPS+si-TIRAP group were significantly lower than those in the control group.Conclusion:The increased expression of TIRAP in epithelial ovarian cancer cells may be one of the mechanisms of epithelial ovarian cancer proliferation.Down-regulation of TIRAP expression can inhibit epithelial ovarian cancer cell proliferation,promote cell apoptosis and reduce the expression of downstream immune molecules by inhibiting the activation of NF-κB pathway.
作者
王杰
钱景荣
卢莹莹
李文辉
WANG Jie;QIAN Jingrong;LU Yingying;LI Wenhui(Clinical Laboratory,Harbin Medical University Cancer Hospital,Heilongjiang Harbin 150081,China)
出处
《现代肿瘤医学》
CAS
北大核心
2021年第22期3908-3913,共6页
Journal of Modern Oncology
作者简介
王杰(1992—),女,山东济南人,检验师,硕士研究生,主要从事临床检验诊断相关研究,E-mail:1048973913@qq.com;通讯作者:李文辉(1973—),女,黑龙江哈尔滨人,主任医师,硕士生导师,主要从事临床检验诊断相关研究,E-mail:iamliwh@sina.com。
