摘要
目的:研究RNA干扰抑制HMGB1的表达对高侵袭转移能力的卵巢癌细胞亚系S1,A1和HO8910PM增殖和侵袭能力的影响。方法:构建靶向HMGB1基因的慢病毒vshRNA载体,转染具有高侵袭转移能力的S1,A1和HO8910PM,同时利用细胞爬片、real-time RT-PCR、Western blot等方法检测RNA干扰抑制HMGB1表达的效果。绘制生长曲线,软琼脂培养克隆形成实验和Boyden chamber侵袭移动实验检测RNA干扰抑制HMGB1基因表达对卵巢癌细胞增殖和侵袭能力的影响。结果:细胞爬片、real-time RT-PCR和Western blot均证实慢病毒HMGB1 shRNA载体成功抑制HMGB1在S1,A1和HO8910PM中的mRNA和蛋白表达。生长曲线,软琼脂培养克隆形成实验和Boydenchamber侵袭移动试验结果显示HMGB1的表达下调抑制卵巢癌细胞的增殖和侵袭能力。结论:HMGB1的表达下调可明显抑制卵巢癌细胞的体外增殖和侵袭能力,为卵巢癌治疗提供了新思路。
Objective：To study the effects of HMGB1 small interfering RNA（siRNA）on invasion and proliferation of human ovarian cancer cells.Methods：Lentivirus vector with HMGB1 shRNA was constructed and infected the highly invasive ovarian cancer cell line S1,A1 and HO8910PM.After viral infection,more than 80% of cells were GFP-positive.real-time RT-PCR,Western blot and IHC were performed to investigate HMGB1 mRNA and protein expression.Cell proliferation was determined by using MTT assay and anchorage independent soft agar colony formation assay.Cell invasion and migration were determined by cell invasion assay and migration assay using Boyden chamber.Results：real-time RT-PCR,Western blot and IHC results confirmed the down-regulation of HMGB1 expression by its shRNA in the highly invasive cell line S1,A1 and HO8910PM.HMGB1 expression was reduced by 80%~90% at both the mRNA and protein levels.MTT assay and soft agar colony formation assay results showed that HMGB1 knockdown significantly reduced cell proliferation of S1,A1 and HO8910PM.Cell invasion assay and migration assay results showed that HMGB1 knockdown significantly reduced cell invasion and migration of S1,A1 and HO8910PM.Conclusion：Knockdown of HMGB1 expression can significantly suppress ovarian cancer cell proliferation and inhibite cell migration and invasion.HMGB1 may serve as a new therapeutic target for the treatment of ovarian cancer in the future.
出处
《现代妇产科进展》
CSCD
2012年第4期241-245,共5页
Progress in Obstetrics and Gynecology
基金
山东省优秀中青年科学家科研奖励基金(No:2007BS03025)
关键词
HMGB1
卵巢肿瘤
侵袭转移
HMGB1
Ovarian neoplasms
Invasion and metastasis
