Bae S N, Kim J, Lee Y S, Kim J D, Kim M Y, Park L-O
Department of Obstetrics and Gynecology, Division of Gynecologic Oncology, Kangnam St. Mary's Hospital, The Catholic University of Korea, College of Medicine, Seoul 137-040, South Korea.
Placenta. 2007 Jan;28(1):22-30. doi: 10.1016/j.placenta.2006.01.003. Epub 2006 Feb 28.
This study investigated the cytotoxic effect of zinc-citrate compound (CIZAR) on choriocarcinoma cell lines. Primary cultured normal trophoblast cells (NPT), human tumorigenic poorly differentiated trophoblast cell line (HT), and choriocarcinoma cell line (BeWo) were exposed to different concentrations of CIZAR and cultured at different times. Cell viability was determined by CCK-8 assay. The effects on cell cycle progression, population distribution and apoptotic incidence were determined by flow cytometry. The appearance of apoptosis was confirmed by DNA laddering and DAPI staining. The quantitative analysis of telomerase was measured by TRAPeze telomerase detection kit. The molecular mechanism of CIZAR-induced apoptosis was examined with Western blot analysis and colorimetric caspase-3 activity assay. In in vitro condition, CIZAR had a selective cytotoxic effect on choriocarcinoma cell line in dose- and time-dependent patterns. Flow cytometric analysis, DNA laddering, and DAPI staining indicated that BeWo cells only have been induced apoptosis by CIZAR. Shortening of telomere was also observed only in BeWo cells. Results also displayed that CIZAR-induced apoptosis involves the up-regulation of p21(WAF1) and Bax protein and down-regulation of Bcl-2 which were accompanied by the activation of caspase-3. Taken together, our results suggest that CIZAR is an apoptotic inducer in malignant trophoblast cells (BeWo).
本研究调查了柠檬酸锌化合物（CIZAR）对绒毛膜癌细胞系的细胞毒性作用。将原代培养的正常滋养层细胞（NPT）、人致瘤性低分化滋养层细胞系（HT）和绒毛膜癌细胞系（BeWo）暴露于不同浓度的CIZAR，并在不同时间进行培养。通过CCK-8法测定细胞活力。通过流式细胞术测定对细胞周期进程、群体分布和凋亡发生率的影响。通过DNA梯状条带分析和DAPI染色确认凋亡的出现。通过TRAPeze端粒酶检测试剂盒对端粒酶进行定量分析。用蛋白质免疫印迹分析和比色法caspase-3活性测定法检测CIZAR诱导凋亡的分子机制。在体外条件下，CIZAR对绒毛膜癌细胞系具有剂量和时间依赖性的选择性细胞毒性作用。流式细胞术分析、DNA梯状条带分析和DAPI染色表明，BeWo细胞仅被CIZAR诱导凋亡。仅在BeWo细胞中也观察到端粒缩短。结果还显示，CIZAR诱导的凋亡涉及p21（WAF1）和Bax蛋白的上调以及Bcl-2的下调，同时伴有caspase-3的激活。综上所述，我们的结果表明CIZAR是恶性滋养层细胞（BeWo）中的一种凋亡诱导剂。
