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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 investigate the inhibitory effect of 15d-PGJ2 and PTEN plasmid transfection on human breast cancer cell line MCF-7 cultured in vitro. Methods MCF-7 cells were seeded into 96-well plates, and 4 experimental groups were set up according to 2 × 2 factorial design. The transfected group was transfected with 1μg pcDNA3.0-PTEN plasmid into MCF-7 cells by lipofectamine. The combination group was given pcDNA3.0-PTEN transfection and peroxisome proliferator-activated receptor (PPAR) γ The natural agonist 15d-PGJ2 40μmol / L; intervention group 15d-PGJ2 40μmol / L treatment cells; control group conventional cultured MCF-7 cells without any treatment. The inhibitory effects of 15d-PGJ2 and plasmid transfection on the growth of MCF-7 cells were observed by MTT assay. The changes of cell cycle and apoptosis were detected by flow cytometry. Results The results of MTT assay showed that compared with the control group, the transfection group and the intervention group could effectively inhibit the growth of MCF-7 cells. At 48, 72 and 96h, the combined effect was more obvious than that of the other two groups (P <0.05). The cells in the control group showed no growth inhibition at all time points. The cell inhibitory rates in the transfected group, combined group and the intervention group were 2%, 0% and 0% at 24 hours, 28% at 48 hours, 39% and 26% respectively; at 72h, they were 74%, 84% and 70% respectively; at 96h, they were 85%, 89% and 80% respectively. The results of cell cycle assay showed that the transfection group had a stronger effect of blocking cells in G0 and G1 phase (P <0.05) and the effect of intervention group was weaker (P <0.05) compared with the control group. The blocking effect of G0 and G1 phase in combined group was weaker than that in transfected group (P <0.05), but it was stronger than that in intervention group (P <0.05). Apoptosis detection showed that compared with the control group, the transfection group, the intervention group and the combination group could not effectively induce the apoptosis of MCF-7 cells (P <0.05). Conclusions PTEN gene transfection and target drug 15d-PGJ2 combined with MCF-7 can effectively inhibit cell growth but not apoptosis.