摘要
目的:探讨敲降人附睾蛋白4(HE4)和配对盒基因8抗原(PAX8)基因后对紫杉醇药+铂类药(TC方案)治疗的上皮性卵巢癌OVCAR3细胞增殖、迁移、侵袭及凋亡的影响。方法:分别设计和合成敲降HE4和PAX8的单靶siRNA(HE4-siRNA或PAX8-siRNA)及双靶siRNA(HE4+PAX8-siRNA)和阴性siRNA序列,并与质粒载体pGCsi-H1相连形成重组质粒,分别转染人上皮性卵巢癌OVCAR3细胞形成HE4-siRNA组、PAX8-siRNA组、HE4+PAX8-siRNA组和siRNA-NC组,同时设立空白对照组(无任何处理)。用TC方案(紫杉醇3.13μg/ml+卡铂2.82μg/ml,)分别处理上述5组细胞后,用MTT法、划痕愈合实验、Transwell侵袭实验、流式细胞术检测各组细胞的增殖、迁移、侵袭及凋亡能力的变化。结果:敲降HE4和PAX8基因后,与siRNA-NC组和空白对照组比较,HE4-siRNA组、PAX8-siRNA组、HE4+PAX8-siRNA组卵巢癌OVCAR3细胞的增殖、迁移和侵袭能力显著降低(均P<0.01),细胞凋亡率显著升高(P<0.01),尤以同时敲降HE4和PAX8基因的效果更佳。结论:敲降HE4和PAX8基因均可增强TC方案抑制上皮性卵巢癌细胞增殖、迁移及侵袭能力并促进其凋亡,尤以HE4和PAX8基因同时敲除的效果更好。
Objective:To investigate the effect of human epididymal protein 4(HE4)and paired box gene 8(PAX8)gene knockdown on proliferation,migration,invasion and apoptosis of human epithelial ovarian cancer OVCAR3 cells treated with TC regimen(paclitaxel+carboplatin).Methods:Sequences of single-target siRNA(HE4-siRNA or PAX8-siRNA)and double-target siRNA(HE4+PAX8-siRNA)as well as negative siRNA were respectively designed and synthesized,and then linked with plasmid vector pGCsi-H1 to obtain the recombinant plasmids.The obtained recombinant plasmids were then transfected into human epithelial ovarian cancer OVCAR3 cells,namely HE4-siRNA group,PAX8-siRNA group,HE4+PAX8-siRNA group and siRNA-NC group,respectively.The blank control group was also set up(without any treatment).The cells in above five groups were treated with TC regimen(paclitaxel 3.13 g/ml+carboplatin 2.82μg/ml),and the changes in proliferation,migration,invasion and apoptosis of the cells were detected by MTT,wound-healing assay,Transwell chamber assay,and flow cytometry,respectively.Results:After knocking down the HE4 and PAX8 genes,compared with siRNA-NC group and blank control group,the proliferation,migration and invasion abilities of OVCAR3 cells in HE4-siRNA group,PAX8-siRNA group and HE4+PAX8-siRNA group significantly decreased(all P<0.01),and the apoptosis rate significantly increased(P<0.01),especially in HE4+PAX8-siRNA group.Conclusion:Knockout of either HE4 or PAX8 can enhance the effect of TC regimen on inhibiting proliferation,migration and invasion as well as promoting apoptosis of epithelial ovarian cancer cells,and the effect of simultaneous down-regulation of HE4 together with PAX8 is better.
作者
唐海峰
李贤
刘蓓
TANG Haifeng;LI Xian;LIU Bei(Department of Gynaecology,Shenzhen Hospital,the University of Hong Kong,Shenzhen 518055,Guangdong,China)
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2019年第7期757-761,共5页
Chinese Journal of Cancer Biotherapy
基金
广东省深圳市知识创新计划—基础研究项目(No.20140331130511)~~
关键词
上皮性卵巢癌
OVCAR3细胞
人附睾蛋白4
配对盒基因8抗原
TC方案
增殖
迁移
侵袭
凋亡
epithelial ovarian cancer
OVCAR3 cell
human epididymal protein 4(HE4)
paired box gene 8(PAX8)
TC regimen
proliferation
migration
invasion
apoptosis
