大蒜素对人结肠癌HT29细胞增殖影响及作用机制研究_临床医学论文.docVIP

大蒜素对人结肠癌HT29细胞增殖影响及作用机制研究_临床医学论文 大蒜素对人结肠癌HT29细胞增殖影响及作用机制研究_临床医学论文 作者：王旭平　赵玲　王容 郭锋 辛华【摘要】 　　目的：探讨大蒜素对体外培养结肠癌HT29细胞增殖的影响，并探讨其作用机制。方法：采用四甲基偶氮唑蓝（MTT）法测定不同浓度大蒜素对HT29细胞增殖抑制率；流式细胞术测定细胞周期变化及凋亡相关蛋白Bcl-2与Bax的表达。结果：MTT显示大蒜素能抑制人结肠癌HT29细胞的增殖，并具有时间-剂量依赖性；流式细胞术测定结果显示10 μg/ml大蒜素可明显诱导HT29细胞凋亡，并将细胞周期阻滞在G2/M期；可降低癌基因蛋白Bcl表达，而促进Bax表达。结论：大蒜素对人结肠癌HT29细胞增殖具有抑制作用，可能与细胞周期阻滞和凋亡相关蛋白表达有关。 【关键词】 大蒜素　结肠癌细胞系T29　凋亡　流式细胞术 【ABSTRACT】 Objective: To study the affection of Diallyl trisulfide on cultured human colon carcinoma cell line HT-29 and its mechanism. Methods: The viability of cells after various treatments were determined using the method of MTT. Flow cytometry were used to analyze the change of cell cycle and Bcl-2, Bax levels. Results: The proliferation of human colon carcinoma HT-29 cells were inhibited by Diallyl trisulfide. The inhibition was associated with dose and time dependence. Flow cytometry results showed that: 1） When cells were treated with Diallyl trisulfide at the concentration of 10 μg/mL for 12 h and 24 h, the percentage of G0/G1 phase cells was decreased and that of G2/M phase cells was significantly increased compared with those in the control group.2） Diallyl trisulfide down regulated Bcl expression, while up-regulated bax expression. Conclusions: It was suggested that Diallyl trisulfide inhibited HT29 cell proliferation. And these were associated with arrest of cell cycles and down regulation of Bcl2/Bax ratio. 【KEY WORDS】 Diallyl trisulfide·Human colon carcinoma cell line HT-29·Apoptosis·Flow cytometry 大蒜素是一种从大蒜鳞茎中分离出的具有多种生物学活性的化合物，其化学名为二烯丙基三硫化物。现代医学研究证实大蒜素具有抗菌、消炎、提高机体免疫力和抗肿瘤的作用［1-3］。体外研究显示,大蒜素可以通过诱导癌细胞（如胃癌、前列腺癌）生成活性氧（ROS），上调Bax，Bak和激活Caspase 3等方式促进其凋亡［1，4，5］，但大蒜素对结肠癌作用的研究较少。本研究以体外培养的结肠癌HT29细胞为材料，探讨大蒜素对结肠癌细胞增殖的影响并探讨其机制，以期为结肠癌的防治提供实验依据。 1　材料与方法 1.1　材料　人结肠癌HT29细胞由山东大学医学院细胞生物学研究所提供；大蒜素（二烯丙基代磺酸酯）为上海禾丰制药有限公司生产；MTT、二甲基亚砜（DMSO）、Ho33342、碘化丙啶（PI）为美国Sigma产品；DMEM、新生牛血清为美国Gibco产品；Bcl-2、Bax单克隆抗体购自美国BD公司。 1.2　方法 1.2.1　细胞培养　人结肠癌HT29细胞用含10%小牛血清的DMEM培养液，于37 ℃、5%CO2饱和湿度条件下静置培养。选择对