重组蛋白LHRH-PE40对卵巢癌细胞增殖与凋亡的影响

摘要

Objective;To study the effects of recombinant protein LHRH-PE40 on ovarian cancer cell proliferation and apoptosis. Methods proliferation inhibition assay was taken in vitro cultured ovarian cancer SK cell. Different concentration of LHRH-PE40 was added into cell culture solution. The effects of LHRH-PE40 on ovarian cancer cell proliferation were analyzed by enzyme micro-plate reader 1-4 days after adding LHRH-PE40. The effects of LHRH-PE40 on proliferative factor Ki-67, anti-apoptosis gene Bcl-2 and cell cycle in ovarian cancer SK cells were detected by immunohistochemical staining and Cytometry. Results: LHRH-PE40 could inhibit the growth of in vitro cultured epithelial ovarian cancer SK cell, which presented time and concentration dependency. The best action time was 48-72 hours after administration of drugs. Under the concentration of medicine ipo/ml, the positive cell rate of Ki-67 (the rate before using medication was 73%, after was 25%) and Bcl-2 (the rate before using medication was 92 %, after was 43%) was decreased after medication, and the ratio of apoptosis was increased (from 0.8% to 3.0%). Under 0.5 ng/ml concentration, the cell ratio of Go/G1 was also increased, in which the cell ratio of S phase was increased and the ratio of G2/M decreased. Conclusions:LHRH-PE40 can kill ovarian cancer cells and promote apoptosis of ovarian cancer cell.%目的:研究重组蛋白LHRH-PE40对卵巢癌细胞增殖及凋亡的影响.方法:采用体外培养的人卵巢癌SK细胞进行细胞增殖抑制实验,在培养液中加入不同浓度的LHRH-PE40,于加药后1～4天用酶标分析仪分析LHRH-PE40对卵巢癌细胞增殖的影响.用免疫组化方法及流式细胞仪检测LHRH-PE40对卵巢癌SK细胞的增殖相关因子Ki-67、抗凋亡基因Bcl-2及细胞周期的影响.结果:LHRH-PE40可以有效抑制体外培养卵巢癌SK细胞的生长,呈时间和浓度的依赖性.最佳作用时间为加药后48～72小时.在药物浓度为1μg/ml的条件下,用药后癌细胞中的Ki-67(用药前73％,用药后25％)及Bcl-2(用药前92％,用药后43％)阳性细胞比率均下降.凋亡细胞比率增加(由用药前0.8％增加到用药后3.6％).在0.5 μg/ml浓度下,细胞的G0/G1期细胞比率增加；S期细胞比率增加,而G2/M期减少.结论:LHRH-PE40可杀灭卵巢癌细胞及促进卵巢癌细胞凋亡.