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孕激素对人卵巢癌细胞株HO8910细胞增殖及凋亡的影响 |
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ion,expression of intracellular bcl-2 protein was analyzed by flow cytometric indirect immunofluorescent technique. Results Progesterone of 1×10-7～1×10-5mol/L inhibited HO8910 cell growth significantly in a dose-dependent manner （P<0.01）. After treatment with progesterone, the enhanced G0/G1 arrest was accompanied with the enhanced apoptotic peak and percentage, as well apoptotic cells were found more than those in control group (P<0.05). By light and electron microscopy, there were many morphological characteristics of apoptosis including compaction and margination of nuclear chromatin, nuclear fragments, and apoptotic bodies. Analysis on expression of intracellular bcl-2 protein showed that progesterone could down-regulate bcl-2 protein and at concentration of 1×10-5mol/L it could almost block bcl-2 expression. Conclusions It is suggested in the present study that progesterone can inhibit the proliferation of epithelial ovarian cancer cells in vitro and there is an accordant dose-response relationship. Its anticancer effect seems to be due to induction of apoptosis which maybe a result of down-regulation of the anti-apoptotic protein bcl-2.
【Key words】 Ovarian neoplasms; Progesterone; Cystadenocarcinoma, serous; Apoptosis; Tumor cells, cultured
细胞凋亡受阻或缺陷是形成肿瘤的机理之一，设法促进细胞凋亡是治疗肿瘤的方向。细胞凋亡和细胞增殖一样，可受到某些生长因子和激素的调节［1］。卵巢癌的激素疗法是否通过诱导细胞凋亡而起作用，目前国内外报道很少。本研究采用多种检测手段，观察孕激素(P)对卵巢癌细胞株HO8910细胞体外增殖及凋亡的影响，以期寻找治疗卵巢癌的有效的辅助措施。
材料与方法
一、材料
卵巢癌细胞株HO8910来源于人卵巢浆液性囊腺癌，由中国科学院上海细胞生物所提供。P和四甲基偶氮唑蓝（MTT）均为美国Sigma公司产品。 细胞凋亡检测试剂盒终末脱氧核苷酸转移酶介导的dUTP缺口末端标记法(TUNEL)为德国宝灵曼公司产品。鼠抗人bcl-2 单克隆抗体、异硫氰酸荧光素（FITC）标记羊抗鼠IgG抗体购自北京中山生物技术公司。
二、方法
1.细胞的培养:HO8910细胞培养方法参见文献［2］。
2.细胞增殖的测定：按上述培养方法得到的HO8910单细胞悬液，以每孔6×103个细胞加入96孔培养板中，置于37℃、5％二氧化碳培养箱中培养，第2天待大部分细胞贴壁后4℃培养1 h，以促成细胞同步化生长［2］；换培养液，实验组加入不同浓度的P，每孔20 μl，使其终浓度分别为1×10-8、1×10-7、1×10-6、1×10-5mol/L，每一浓度均设8个平行孔，对照组(下同)加入20 μl空白培养液。继上一页 [1] [2] [3] 下一页
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