ERCC1反义寡核苷酸对人卵巢癌细胞增殖、周期、药物敏感性及侵袭能力的影响  

Effects of ERCC1 Antisense oligonucleotide on proliferation,cell cycle,and drug sensitivity of human ovarian cancer cell SKOV3/DDP

作　　者：周英琼[1] 梁梦[2] 黎雁英[1] 李运千[1] 陆竞艳[1] 田佳[1] 

机构地区：[1]桂林医学院附属医院病理科,广西桂林541001 [2]浙江中医药大学附属第三医院病理科,浙江杭州310005

出　　处：《中国现代医学杂志》2013年第29期24-29,共6页China Journal of Modern Medicine

基　　金：2008年桂林医学院硕士生导师专项课题(No:院科字[2008]8号);2010年度广西教育厅科研资助项目(No:201012MS173)

摘　　要：目的探讨体外ERCC1反义寡核苷酸(ERCC1-ASODNs)对人卵巢癌细胞耐药株SKOV3/DDP生长周期、增殖及侵袭力改变的影响及其逆转顺铂耐药的机制。方法将设计合成的特异性ERCC1-ASODNs片段以及无义寡核苷酸序列(NSODNs)转染至SKOV3/DDP细胞后实验分组,采用流式细胞学技术(FACS)检测细胞生长周期,MTT法检测转染前后细胞的生长抑制情况及对顺铂敏感性的差异,应用RT-PCR法检测各组细胞ERCC1 mRNA的表达情况;Transwell小室实验检测转染前后其侵袭力的改变。结果 ERCC1-ASODNs可下调SKOV3/DDP细胞的ERCC1基因的表达;抑制其细胞的生长;促使其细胞周期发生改变,S期细胞增多,G0/G1期细胞减少;转染后SKOV3/DDP细胞的侵袭力减弱,穿过基质膜细胞数目减少。结论运用ERCC1反义寡核苷酸能特异、高效抑制靶基因ERCC1的表达;且减弱卵巢癌耐药细胞的增殖及侵袭能力,增强其对顺铂的敏感性。[Objective] To explore the ERCC1 Antisense oligonucleotide on proliferation, cell cycle, and drug sensitivity of human ovarian cancer cell line SKOV3/DDP. [Methods] ERCC1 Antisense oligonucleotide was transfected into SKOV3/DDP cells by transfection reagent X-tremeGENE siRNA transfection reagent. Non- sense oligonucleotide sequence group, untransfected group and transfection reagent were used as control groups. The cell cycle distribution was analyzed by flow cytometry. The proliferation of SKOV3/DDP cells was examined by MTY assay. The expression of ERCC1 mRNA was identified by RT-PCR. Cisplatin （DDP） sensi- tivity experiment was implemented on SKOV3/DDP cells with ERCC1 Antisense oligonucleotide. [Results] The Antisense oligonucleotide can knockdown the expression of ERCC1 in SKOV3/DDP cells obviously, retard the cells in S phase, inhibit the cell＇s proliferation, and weaken its invasion force. The IC50 of DDP in SKOV3/ DDP cells which transfected with ERCC1 antisense oligonucleotide was decreased. [Conclusion] ERCC1 Anti- sense oligonucleotide can partly suppress the expression of ERCC1 in SKOV3/DDP cells, inhibit the cell proliferation, and retard the cells cycle which implies that ERCC1 Antisense oligonucleotide may partly reverse the drug resistance of ovarian cancer.

关 键 词：卵巢癌 ERCC1 反义寡核苷酸 细胞周期 耐药 

分 类 号：Q253[生物学—细胞生物学] R737.31[医药卫生—肿瘤]