特异siRNA下调卵巢癌H08910细胞NAC-1基因表达并抑制生长

基础医学与临床 ›› 2012, Vol. 32 ›› Issue (10): 1221-1223.

特异siRNA下调卵巢癌H08910细胞NAC-1基因表达并抑制生长

桂玲¹,王静²,祝爱珍²,刘成成²,刘革修²

1. 湖南省肿瘤医院妇瘤科
2. 暨南大学医学院血液研究所

收稿日期:2011-08-30 修回日期:2012-03-07 出版日期:2012-10-05 发布日期:2012-09-28
通讯作者: 刘革修 E-mail:53483119@qq.com

The inhibitory effect of specific small interfering RNA against nucleus accumbens-1 gene on cell proliferation of ovarian cancer cells

Received:2011-08-30 Revised:2012-03-07 Online:2012-10-05 Published:2012-09-28
Contact: Ge-Xiu LIU E-mail:53483119@qq.com

摘要/Abstract

摘要： 目的：研究小RNA干扰NAC-1 (Nucleus accumbens-1，Nac1 or NAC-1)基因表达对卵巢癌HO8910细胞增殖的影响。方法：设计、合成针对NAC-1基因的特异性小干扰RNA（small interfering RNA，siRNA）及阴性siRNA，并同时设立空白对照，然后用脂质体转染HO8910细胞。通过实时荧光定量RT-PCR 和Western印迹法分别检测转染前后HO8910细胞中NAC-1 mRNA和蛋白表达水平，MTT法和克隆形成实验检测细胞增殖率，FCM法检测细胞周期分布。结果：针对NAC-1基因的特异性siRNA均能抑制NAC-1 mRNA和蛋白的表达，其中NAC-1-siRNA-1的沉默效率最高。转染NAC-1-siRNA-1 48h后，HO8910细胞中NAC-1 mRNA和蛋白表达水平分别下调74%和81%，而且细胞增殖明显受到抑制，细胞周期被阻滞于G1期，与转染阴性siRNA 及未转染组比较，有显著性差异(P?0.05)。结论：NAC-1特异性siRNA能够有效沉默NAC-1基因表达，并显著抑制卵巢癌HO8910细胞增殖。

关键词: NAC-1, siRNA, 卵巢癌, 细胞增殖, 细胞周期

Abstract: Objective: To investigate the effect of specific small interfering RNA (siRNA) against nucleus accumbens-1（Nac1 or NAC-1）gene on the proliferation of ovarian cancer cell line HO8910. Methods: Three specific NAC-1 siRNAs and one negative siRNA were individually transfected into HO8910 cells for 48 h . The expression levels of NAC-1 mRNA and protein were analyzed by real-time fluorescence quantitative PCR (qRT-PCR) and Western blotting, respectively. The cell proliferation was determined by MTT assay and plate clone formation assay. The distribution of cell cycle was assessed by flow cytometry. Results: In comparison with the other two kinds of NAC-1 siRNAs, NAC-1-siRNA-1 was demonstrated to have the strongest silencing effect on NAC-1 gene expression in HO8910 cells at 48 h after transfection. The expression levels of NAC-1 mRNA and protein were reduced by 65% and 80%, respectively. The growth rate of HO8910 cells transfected with NAC-1-siRNA-1 was significantly decreased. Moreover, the cell cycle arrest in the G1 phase was observed when HO8910 cells were transfected with NAC-1-siRNA-1 for 48 h. Conclusion: Specific NAC-1 siRNA can effectively inhibit the expression of NAC-1 gene and significantly suppress cell proliferation of ovarian cancer HO8910 cells.

Key words: ovarian cancer, nucleus accumbens-1, small interfering RNA, Cell proliferation, Cell cycle

中图分类号:

R737.14

桂玲王静祝爱珍刘成成刘革修. 特异siRNA下调卵巢癌H08910细胞NAC-1基因表达并抑制生长[J]. 基础医学与临床, 2012, 32(10): 1221-1223.

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