摘要
目的探究mi R-449c-5p对卵巢癌细胞紫杉醇(paclitaxel,PTX)耐药影响及潜在调控机制。方法q RT-PCR检测mi R-449c-5p表达,蛋白免疫印迹分析ABCC1的蛋白表达。将mi R-449c-5p、mi R-NC、ABCC1过表达载体以及pc DNA 3.1(+)载体转染PTX耐药的卵巢癌细胞A2780/PTX和SKOV3/PTX,利用CCK-8法以及transwell实验分析细胞表型变化。双荧光素酶报告实验鉴定mi R-449c-5p和ABCC1之间的联系。移植瘤鼠模型实验体内分析mi R-449c-5p和PTX联合处理对肿瘤生长的影响。结果与HOSEPi C细胞相比,mi R-449c-5p表达在A2780和SKOV3细胞中较低(P<0.05),在A2780/PTX和SKOV3/PTX细胞中最低(P<0.05)。mi R-449c-5p在A2780/PTX和SKOV3/PTX细胞中靶向ABCC1。与mi R-NC转染组相比,mi R-449c-5p转染组中IC50值降低、侵袭细胞数减少(P<0.05);和mi R-449c-5p和pc DNA共转染组比较,mi R-449c-5p和ABCC1共转染组的PTX的IC50值显著增加、侵袭细胞数增加(P<0.05)。和mi R-NC+PBS组比较,鼠模型移植瘤体积在mi R-NC+PTX组和mi R-449c-5p+PBS组显著降低(P<0.05),与mi R-NC+PTX组比较,移植瘤体积在mi R-449c-5p+PTX组显著降低(P<0.05)。结论Mi R-449c-5p通过调控ABCC1提高卵巢癌对PTX的敏感性。
Objective To explore the effects of miR-449c-5p on the paclitaxel(PTX)sensitivity of PTX-resistant ovarian cancer cells and the potential regulatory mechanism.Methods MiR-449c-5p expression was analyzed by qRTPCR,and ABCC1 protein expression was detected by western blotting assay.PTX-resistant ovarian cancer cells including A2780/PTX and SKOV3/PTX were transfected with miR-449c-5p,miR-NC,ABCC1 overexpression vector and pcDNA 3.1(+)vector,and then cell phenotypic changes were analyzed by CCK-8 assay and transwell assay.Dual-luciferase reporter assay was used to identify the association between miR-449c-5p and ABCC1.The effect of combination treatment of miR-449c-5p and PTX on tumor growth was analyzed in vivo by a xenograft mouse model assay.Results Compared with HOSEPiC cells,miR-449c-5p expression was lower in A2780 and SKOV3 cells(P<0.05),and was the lowest in A2780/PTX and SKOV3/PTX cells(P<0.05).MiR-449c-5p interacted with ABCC1 in A2780/PTX and SKOV3/PTX cells.Compared with the miR-NC transfection group,the miR-449c-5p transfection group showed reduced IC50 value of PTX and the decreased number of invasive cells(P<0.05).Compared with the miR-449c-5p and pcDNA co-transfection group,the IC50 value of PTX and the number of invasive cells in the miR-449c-5p and ABCC1 co-transfection group was significantly increased(P<0.05).Compared with the miR-NC+PBS group,the transplanted tumor volume in the mouse model were significantly decreased in the miR-NC+PTX group and the miR-449c-5p+PBS group(P<0.05),and compared with the miR-NC+PTX group,the transplanted tumor volume was significantly decreased in the miR-449c-5p+PTX group(P<0.05).Conclusion MiR-449c-5p enhanced the sensitivity of ovarian cancer to PTX by interacting with ABCC1.
作者
高玉霞
王文翔
李章欢
刘宇
李力
何乾
韩琛
GAO Yuxia;WANG Wenxiang;LI Zhanghuan;LIU Yu;LI Li;HE Qian;HAN Chen(Department of Gynecology and Oncology,Xinxiang Central Hospital,Fourth Clinical College,Xinxiang Medical College,Xinxiang Henan 453000,China;Obstetrics Department,Xinxiang Central Hospital,Fourth Clinical College,Xinxiang Medical College,Xinxiang Henan 453000,China)
出处
《中国妇产科临床杂志》
CSCD
北大核心
2024年第5期404-407,共4页
Chinese Journal of Clinical Obstetrics and Gynecology
基金
河南省医学科技攻关计划联合共建项目(LHGJ20210914)
新乡市科技攻关项目(GG2020041)。
