贵州医科大学学报

2015, v.40;No.183(12) 1325-1330

[打印本页] [关闭]
本期目录(Current Issue) | 过刊浏览(Past Issue) | 高级检索(Advanced Search)

SiRNA沉默cdc2基因逆转卵巢癌顺铂耐药的实验研究
Reversal of Cisplatin Resistance in Human Ovarian Carcinoma Cells by RNAi Silencing of cdc2 Gene

任婕;彭芝兰;杨英捷;肖子文;
REN Jie;PENG Zhilan;YANG Yingjie;XIAO Ziwen;Department of Obstrics and Gynecology,Affiliated hospital of Guizhou medical University;Department of Obstetrics and Gynecology,Second Hospital of West China Medical Center of Sichuan University;

摘要(Abstract):

目的:探讨利用RNAi沉默cdc2基因逆转人卵巢上皮性癌细胞顺铂耐药的可行性,为卵巢癌顺铂耐药逆转提供一个可选择的靶点。方法:针对cdc2基因的DNA设计具有特异性的SiRNA,经脂质体转染卵巢癌顺铂耐药细胞A2780/DDP,采用逆转录-聚合酶链式反应(RT-PCR)和免疫印迹法检测转染组cdc2基因表达是否抑制;用MTT法测定转染前后A2780/DDP细胞对顺铂敏感性的变化,流式细胞仪检测细胞凋亡率。结果:(1)转染cdc2-SiRNA在RNA及蛋白水平均有效抑制A2780/DDP细胞中cdc2的表达,抑制率分别为78.2%、76.8%;(2)顺铂化疗IC50,A2780/DDP-S(0.66±0.02)较A2780/DDP(5.02±0.05)显著下降(P<0.01);(3)5 mg/L顺铂作用后,A2780/DDP-S与A2780/DDP比较,各时间点细胞生长抑制率明显增加(P<0.05),细胞凋亡率明显增多(P<0.05)。结论:在A2780/DDP细胞中,SiRNA能有效抑制cdc2基因的表达,并能部分恢复其对顺铂化疗的敏感性。
Objective: To investigate the feasibility of RNAi silencing of the cdc2 gene in human ovarian carcinoma cell line A2780 / DDP to reverse the cisplatin resistancein,and provide a novel therapy target for ovarian chemotherapy resistance. Methods: The specific SiRNA was synthesized for cdc2 and transfected into human ovarian carcinoma cisplatin resistance cell line A2780 / DDP by liposome.RT-PCR and cell immunostaining were used to detect the changes of cdc2 mRNA and protein in A2780 / DDP cell. The drug sensitivity to cisplatin was examined by MTT assay. Apoptosis rates were detected by flow cytometry. Results:( 1) The transfection of cdc2-SiRNA could effectively inhibit the expression of cdc2 RNA and protein in A2780 / DDP cells. The inhibition rate was 78. 2% and 76. 8%respectively.( 2) The IC50 of A2780 / DDP-S( 0. 66 ± 0. 02) was significantly decreased compared with A2780 / DDP( 5. 02 + 0. 05).( 3) After treated by cisplatin( 5 mg / L),the cell growth inhibition rate and apoptosis rate of A2780 / DDP-S was significantly increased than that of A2780 / DDP at each time point( P < 0. 05). Conclusion: The SiRNA could effectively block the cdc2 gene expression in A2780 / DDP cell,thereby reversed resistance to cisplatin in some part.

关键词(KeyWords): 人卵巢癌;RNA干扰;cdc2;顺铂耐药;免疫印迹法;逆转录-聚合酶链式反应
human ovarian carcinoma;RNA interference;cdc2;cisplatin resistance

Abstract:

Keywords:

基金项目(Foundation): 贵州省科技厅资助项目(No:E2009-39)

作者(Author): 任婕;彭芝兰;杨英捷;肖子文;
REN Jie;PENG Zhilan;YANG Yingjie;XIAO Ziwen;Department of Obstrics and Gynecology,Affiliated hospital of Guizhou medical University;Department of Obstetrics and Gynecology,Second Hospital of West China Medical Center of Sichuan University;

Email:

DOI: 10.19367/j.cnki.1000-2707.2015.12.010

参考文献(References):

文章评论(Comment):

序号(No.) 时间(Time) 反馈人(User) 邮箱(Email) 标题(Title) 内容(Content)
反馈人(User) 邮箱地址(Email)
反馈标题(Title)
反馈内容(Content)
扩展功能
本文信息
服务与反馈
本文关键词相关文章
本文作者相关文章
中国知网
分享