克霉唑诱导人结肠癌细胞CCL229凋亡的实验研究

Clotrimazole Induced Apoptosis in Human Colon Cancer Cell Line CCL229

背景与目的:国外有研究表明克霉唑可通过持续耗竭细胞内钙库来抑制肿瘤细胞生长。本研究旨在研究克霉唑诱导人结肠癌细胞凋亡的作用。方法:将不同浓度的克霉唑作用于体外培养的人结肠癌CCL229细胞。利用荧光显微镜、电子显微镜观察细胞的形态变化,流式细胞仪检测细胞周期变化及亚二倍体比例,并利用TUNEL法进行凋亡的分子生物学检测。结果:25～35μmol/L的克霉唑均可诱导CCL229细胞凋亡,其诱导作用表现为时间和剂量依赖性。荧光显微镜及电镜下观察到凋亡小体形成等典型的形态学改变。DNA直方图上显示亚二倍体峰,亚二倍体比例随剂量及时间的增加而增加。相应地,G1期细胞比例下降。TUNEL法观察到棕褐色着染的凋亡阳性细胞。结论:克霉唑对人结肠癌细胞系CCL229有明显的凋亡诱导作用,并且能特异地诱导G1期细胞凋亡。

Background &Objective:It was reported that clotrimazole could inhibit the growth of tumor cell by exhausting cellular calcium. This study was designed to investigate whether clotrimazole(CLT)caninduce apoptosis invitro inhumancolon cancer cell line CCL229. Methods:The fluorescent microscope,electron microscopes,flow cytometry and terminal deoxynucleotidyl transferase mediated dUTP biotin nick end labeling(TUNEL)were applied to detect the morphological changes,cells cycle distribution,and DNA fragmentation. Results:Apoptosis can be induced in colon cancer cell CCL229 by CLT at the concentration of 25～35 μmol/L in a dose and time dependent manner. Typical morphological changes of apoptosis such as apoptic body were found under fluorescent microscope and electron microscopes.DNA histograms showed the hypodiploid peak,the ratio of hypodiploid cells was increased with the increase of dose and elongation of time.The ration of cells in G1 phase decrease accordinly. Brown colored positive apoptotic cells were observed with TUNEL method. Conclusion:CLT can induce apoptosis in vitro in human colon cancer cell line CCL 229.The apoptosis inducing effect may be G1 phase specific.