摘要
目的探讨15d-PGJ2和PTEN质粒转染对体外培养的人乳腺癌细胞株MCF-7的抑制作用。方法MCF-7细胞接种96孔板,按2×2析因设计设置4个试验组。转染组用脂质体转染法将pcDNA3.0-PTEN质粒1μg转染MCF-7细胞;联合组给予pcDNA3.0-PTEN转染和过氧化物酶体增殖物激活受体(PPAR)γ天然激动剂15d-PGJ2 40μmol/L;干预组15d-PGJ2 40μmol/L处理细胞;对照组常规培养的MCF-7细胞不做任何处理。采用MTT法观察15d-PGJ2和质粒转染对MCF-7细胞的生长抑制作用;用流式细胞仪检测细胞周期和细胞凋亡的变化。结果MTT法检测细胞吸光度(A)值结果显示,与对照组相比,转染组、干预组均能有效地抑制MCF-7细胞生长。在48、72、96h时点,联用组的效果较其他两组的抑制作用更明显(P<0.05)。对照组细胞在各时间点均未显示出生长抑制;24h时点转染组、联合组、干预组的细胞抑制率分别为2%、0%、0%;在48h时点分别为28%、39%、26%;在72h时点分别为74%、84%、70%;在96h时点分别为85%、89%、80%。细胞周期检测结果显示,与对照组比较,转染组对G0、G1期细胞的阻滞作用较强(P<0.05),干预组作用较弱(P<0.05)。联合组的G0、G1期阻滞效应较转染组弱(P<0.05),但较干预组强(P<0.05)。细胞凋亡检测显示,与对照组相比,转染组、干预组和联合组均不能有效地诱导MCF-7细胞凋亡(P<0.05)。结论PTEN基因转染和靶向药物15d-PGJ2联合作用于体外培养的乳腺癌细胞株MCF-7,能有效抑制细胞生长,但并不诱导细胞凋亡。
Objective To explore the inhibitory effects of 15d-PGJ2 and transfection of PTEN ptasmid on the human breast carcinoma cell line MCF-7 in vitro. Methods MCF-7 cells were seeded on a 96-well plate in the concentration of 5.0×10^6/L. According to the factorial design the cells were assigned into 4 groups.1） Transfection group： MCF-7 calls were transfected with 1μg of pcDNA3.0-PTEN plasmid;2） Combination group： MCF-7 cells were transfected with pcDNA3.0-PTEN plasmid and treated with 40μmol/L 15d-PGJ2, an activator of γ-receptor activated by peroxisome prollferator （PPAR-γ）;3） Interference group： MCF-7 cells were treated with 40μmol/L 15d-PGJ2; and 4） Normal group： normal cells. MTT assay was used to assess the inhibitory effects of 15d PGJ2 and transfection of pcDNA3.0-PTEN on the growth of MCF-7 cells. Flow cytometry was employed to determine the cell cycle and apoptosis. Results It was revealed by the absorbent value （A） determined with MTT that, compared with the normal group, the growth of MCF-7 cells were efficiently suppressed in transfection group and interference group, and the suppression was even more evident in combination group at the time points of 48,72 and 96h （P〈0.05）. In view of the inhibitory rate calculated with A value, the cell growth in normal group was not suppressed at all the time points. At 24h time point, the inhibitory rates in transfection group, combination group and interference group were 2%, 0% and 0%; at 48h time points, it was 28%, 39% and 26%, respectively, and at 72h time points, it was 74%, 84% and 70%, respectively, and at 96h time points, it was 85%, 89% and 80%, respectively. It was shown by the cell cycle detection that, compared with normal group, the growth of more cells was blocked in transfection group, and less cells in interference group in G0 and G1 stages （P〈0.05）. In the combination group, the blocking effect on the cell growth at G0 and G1 stage was less marked compared with that in transfection group （P〈0.05）, but wa
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2008年第12期1434-1437,共4页
Medical Journal of Chinese People's Liberation Army
基金
新疆少数民族科技人才特殊培养计划科研项目(200823114)
新疆维吾尔自治区教育厅高校青年教师启动基金项目(XJEDU2006525)
