自噬基因Beclin1对上皮性卵巢癌细胞SKOV3生长的抑制作用

Growth inhibitory effects of Beclin1 gene on epithelial ovarian cancer SKOV3 cells

目的观察自噬基因Beclin1在人卵巢癌细胞株SKOV3中过表达对肿瘤细胞在体内、体外生长活性的影响。方法脂质体法将重组质粒pcDNA3.1/Beclin1转染人卵巢癌细胞株SKOV3,MTT法分析外源性Beclin1过表达对SKOV3增殖的影响,流式细胞仪检测转染后肿瘤细胞的凋亡及自噬情况。将重组质粒pcDNA3.1/Beclin1转染SKOV3细胞后,接种到裸鼠皮下,观察体内致瘤活性及生长情况;免疫组化检测瘤组织Beclin1蛋白的表达。结果Beclin1在SKOV3中的过表达后抑制肿瘤细胞在体外的生长,抑制率为58.68%;pcDNA3.1/Beclin1转染后SKOV3的凋亡率为(21.26±3.89)%,高于空质粒pcDNA3.1转染组和未转染组,差异有显著性(P<0.05);流式细胞仪检测pcDNA3.1/Beclin1转染后SKOV3细胞MDC平均荧光强度高于pcDNA3.1转染组和未转染组。重组质粒pcDNA3.1/Beclin1使SKOV3细胞在裸鼠体内成瘤时间延长,瘤体体积及质量明显小于空质粒组和空白对照组,抑瘤率为50.27%。结论自噬基因Beclin1在人卵巢癌细胞SKOV3中过表达可抑制SKOV3在体内、体外的增殖,针对自噬基因Beclin1的卵巢癌基因治疗可能具有可行性。

Objective To explore the effect of Beclin1 overexpression on the growth of ovarian carcinoma cell line SKOV3 in vitro and in vivo.Methods The recombinant plasmid pcDNA3.1/Beclin1 was constructed and transfected into SKOV3 cells via lipofectamine 2000.MTT assay was used to evaluate the effect of Beclin1 overexpression on the proliferation and growth of the transfected cells,whose apoptosis and autophagy were analyzed by flow cytometry.SKOV3 cells transfected with the plasmids pcDNA3.1/Beclin1 or pcDNA3.1 were inoculated subcutaneously in nude mice,and their carcinogenic and growth activities in vivo were evaluated.Results MTT assay showed that transfection with pcDNA3.1/Beclin1 significantly inhibited the proliferations of SKOV3 cells,with a cell inhibition rate of 58.68% （P0.05）.The transfection also resulted in a cell apoptosis rate of （21.26±3.89）%,significantly higher than that of pcDNA3.1 trasnfection （P0.05）.Flow cytomerty showed that pcDNA3.1/Beclin1 transfection of SKOV3 cells produced a significantly higher MDC fluorescent intensity than pcDNA3.1 transfection.The SKOV3 cells transfected with vector pcDNA3.1/Beclin1 also showed decreased carcinogenic activity in nude mice,with a growth inhibition rate of 50.27%.Conclusion Beclin1 overexpression can inhibit the proliferation and growth of SKOV3 cells in vitro and vivo,suggesting its potential role in gene therapy of ovarian carcinoma.