Duan Zhen-ling, Peng Zhi-lan, Wang Zan-hong
Department of Obstetrics and Gynecology, First Affiliated Hospital of Kunming Medical College, Kunming 650032, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2010 Apr;30(4):672-6.
To explore the effect of Beclin1 overexpression on the growth of ovarian carcinoma cell line SKOV3 in vitro and in vivo.
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.
MTT assay showed that transfection with pcDNA3.1/Beclin1 significantly inhibited the proliferations of SKOV3 cells, with a cell inhibition rate of 58.68% (P<0.05). The transfection also resulted in a cell apoptosis rate of (21.26-/+3.89)%, significantly higher than that of pcDNA3.1 trasnfection (P<0.05). 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%.
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.
探讨Beclin1过表达对卵巢癌细胞系SKOV3体外及体内生长的影响。
构建重组质粒pcDNA3.1/Beclin1,通过脂质体2000转染入SKOV3细胞。采用MTT法评估Beclin1过表达对转染细胞增殖和生长的影响,通过流式细胞术分析其凋亡和自噬情况。将转染了质粒pcDNA3.1/Beclin1或pcDNA3.1的SKOV3细胞皮下接种于裸鼠,评估其在体内的致癌和生长活性。
MTT法显示,转染pcDNA3.1/Beclin1显著抑制SKOV3细胞的增殖,细胞抑制率为58.68%(P<0.05)。转染还导致细胞凋亡率为(21.26±3.89)%,显著高于pcDNA3.1转染组(P<0.05)。流式细胞术显示,pcDNA3.1/Beclin1转染的SKOV3细胞产生的MDC荧光强度明显高于pcDNA3.1转染组。转染载体pcDNA3.1/Beclin1的SKOV3细胞在裸鼠体内的致癌活性也降低,生长抑制率为50.27%。
Beclin1过表达可抑制SKOV3细胞在体外和体内的增殖和生长,提示其在卵巢癌基因治疗中的潜在作用。