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.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
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)