大蒜素与化疗药物对食管癌细胞EC109疗效比较的实验研究

Antitumor effect of allicin on esophageal cancer EC109 cells compared with chemotherapeutics in vitro

目的:通过与化疗药物比较的实验研究探讨大蒜素对食管癌的抗肿瘤效果及其可能机制。方法:不同浓度的大蒜素、5-氟尿嘧啶和顺铂作用于食管癌细胞EC109,分别在6 h、12 h、24 h、48 h和72 h应用CCK-8法检测EC109细胞的生长抑制率,并以分析细胞乳酸脱氢酶(LDH)活性反映各药物的细胞毒性;Annexin V/PI双染法检测空白对照组、Z-VAD-FMK(caspase-3活性抑制剂)组、大蒜素组、大蒜素+Z-VAD-FMK组、5-氟尿嘧啶组和顺铂组的细胞凋亡情况。分光光度测定法检测细胞内caspase-3、8、9的活性变化。结果:大蒜素呈浓度依赖性及时间依赖性抑制和杀伤食管癌细胞EC109;与5-氟尿嘧啶组和顺铂组比较,大蒜素组LDH活性明显下降;与对照组相比,大蒜素培养细胞内caspase-3、8的活性增强,而细胞内caspase-9的活性变化差异无统计学显著性。结论:大蒜素可能通过活化caspase-8激活外源性凋亡通路,诱导食管癌细胞EC109凋亡,从而抑制肿瘤细胞的生长,且与5-氟尿嘧啶和顺铂相比,大蒜素的细胞毒副作用明显减弱。

AIM：To investigate the inhibitory effect of allicin on human esophageal cancer cell line EC 109 and its possible mechanism by comparison with chemotherapeutic drugs .METHODS：The EC109 cells were treated with different concentrations of allicin , 5-fluorouracil and cisplatin .The cell viability was detected by CCK 8 assay and the activ-ity of lactic dehydrogenase （LDH） was analyzed at 6 h, 12 h, 24 h, 48 h and 72 h.The apoptosis of the EC109 cells in-duced by Z-VAD-FMK, allicin, allicin＋Z-VAD-FMK, 5-fluorouracil and cisplatin was analyzed by flow cytometry with Annexin V-FITC and PI double staining .The enzyme activity changes of caspase-3, caspase-8 and caspase-9 were detected by spectrophotometry .RESULTS：Allicin had inhibitory effect on the growth of the EC 109 cells and killed them in con-centration-dependent and time-dependent manners .LDH activity was decreased compared with 5-fluorouracil and cisplatin . The increased activity of caspase-3 and caspase-8 in allicin-treated cells was statistically significant , but caspase-9 activity changed without statistical significance .CONCLUSION：Allicin inhibits the growth of EC109 cells in concentration-and time-dependent manners through extrinsic apoptosis pathways activated by caspase -8, and the side effects are weaker com-pared with 5-fluorouracil and cisplatin .