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

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.