重组凋亡素真核表达载体诱导人卵巢癌细胞凋亡  

Recombinant Apoptin Eukaryotic Expression Vector Inducing Apoptosis in Human Ovarian Cancer Cells

作　　者：苏海英[1] 曹建国[2] 谢宛玉[1] 

机构地区：[1]南华大学第一附属医院妇产科,湖南衡阳421001 [2]南华大学肿瘤研究所

出　　处：《南华大学学报（医学版）》2005年第4期466-469,共4页Journal of Nanhua University(Medical Edition)

基　　金：湖南省卫生厅课题(编号:Y02-075)

摘　　要：目的观察凋亡素基因(VP3)在卵巢癌细胞株CoC1中诱导凋亡的情况。方法用KpnⅠ、XbaⅠ分别双酶切pMD18-CAVP3和pcDNA3.1(+),分别回收365 bp和5.0 kb大小的片断,然后常规行连接反应,构建重组凋亡素真核表达载体pcDNA3.1-VP3。采用LipofectamineTM2000介导的基因转染法,在体外将pcDNA3.1-VP3转染入卵巢癌细胞株CoC1中,RT-PCR检测凋亡素在细胞中的表达,用流式细胞仪检测细胞凋亡。结果酶切鉴定及测序结果表明重组凋亡素真核表达载体pcDNA3.1-VP3构建成功。RT-PCR证实VP3基因在CoC1中存在并在转录水平表达。转染pcDNA3.1-VP3细胞凋亡率明显高于转染空质粒组(P<0.01)。结论凋亡素可有效诱导卵巢癌细胞CoC1凋亡。Objective To investigate effect of apoptin gene on human ovarian cancer cell （CoCl ）. Methods pMD18 - CAVP3 and pcDNA3.1 （ ＋ ） were respectively double digested by restriction endonuclease Kpn Ⅰ- and Xba Ⅰ , and obtained 365 bp and 5.0 kb fragment, then ligatated them and the recombinant apoptin eukaryotic expression vector pcDNA3.1 - VP3 was constructed. Two plasmids, pcDNA3.1 （ ＋ ） and pcDNA3.1 - VP3, were transfected into human ovarian cancer cells（ CoCl ） by Lipofectamine^TM 2000- mediated gene transfection method in vitro. The expression of apoptin gene at transcription level was proved by RT - PCR. Apoptosis in CoCl was observed by flow cytometry. Results The result of sequencing is consistent with reports in Genbank, apoptin gene was successfully cloned into pcDNA 3.1 （ ＋ ） and recombinant apoptin eukaryotic expression vector pcDNA3.1 - VP3 was successfully constructed. The transient expression of apoptin gene at transcription level had been existed in transfected cells. Apotosis rate in pcDNA3.1 - VP3 - transfected cells was significantly higher than that in pcDNA 3.1 （ ＋ ） - transfected cells. Conclusion Apoptin induces apoptosis in human ovarian cancer CoCl.

关 键 词：鸡贫血病毒 凋亡素 细胞凋亡 卵巢癌 

分 类 号：R737.31[医药卫生—肿瘤] Q786[医药卫生—临床医学]