大蒜素上调TRAIL表达诱导宫颈癌细胞凋亡机制研究

Mechanism research of allicin inducing apoptosis to cervical cancer cells by upregulation of TRAIL

目的观察大蒜素对人宫颈癌Hela、Caski细胞的生长抑制作用,大蒜素作用前后TRAIL及其受体DR4、DR5的表达以及Caspase-3、8、9活性变化,探讨大蒜素诱导宫颈癌细胞凋亡作用及其机制。方法 (1)2009-2010年于河北医科大学第二医院,体外培养Hela、Caski细胞,MTT法检测不同浓度大蒜素对细胞的生长抑制作用。(2)RT-PCR方法检测大蒜素对细胞TRAILmRNA表达的影响。(3)流式细胞仪检测大蒜素作用前后细胞TRAIL及其受体DR4、DR5的表达。(4)检测大蒜素对HeLa、Caski细胞Caspase-3、8、9活性的影响。结果 (1)大蒜素对细胞有不同程度的生长抑制作用,随着药物浓度的增加以及作用时间的延长,细胞毒作用逐渐增强。(2)大蒜素处理细胞12h后TRAILmRNA表达水平明显升高,实验组与对照组比较差异有统计学意义(P<0.05)。(3)大蒜素处理细胞24h后TRAIL蛋白表达水平上调。(4)大蒜素作用48h后,两种细胞半胱氨酸蛋白酶(Caspase)-3、8、9的活性明显增强。结论大蒜素对Hela和Caski细胞具有生长抑制作用;上调TRAIL表达,进而活化Caspase-3、8、9,是大蒜素诱导细胞凋亡的重要机制。

Objective To observe the growth inhibited effect on Hela and Caski cells induced by allicin, and the expressions of TRAIL, DR4, DILS, Caspase-3,8,9 activity before and after allicin treatments. Furthermore to discuss the mechanism of allicin inducing apoptosis to cervical cancer cells. Methods （1）Hela and Caski cells were trained in vitro, to observe the growth inhibitory rates of cells treated by different dose of allicin. （2）Expressions of TRAIL mRNA beibre and ＇after allicin treatment were tested by RT-PCR. （3）Expressions of TRAIL, DR4, DR5 before and after allicin treatment were determined by FCM. （4）Caspase-3,8,9 activity changes before and ＂after allicin treatment were determined. Results （1）Inhibitory rates of Hela and Caski cells after allicin treatments had a time-and-dose-dependent efiect. （2）TRAIL mRNA in Hela and Caski cells ascended markedly after Allicin treatment 12h （P 〈 0. 05 ）. （3）TRAIL expression were up-regulated after Allicin treatment 24h. （4）Caspase-3,8,9 activities were up-regulated obviously ＇after 48h allicin treatment. Conclusion Allicin could inhibit cell proliferation in cervical cancers cell lines of Hela and Caski in vitro. To up-regulate the expression of TRAIL ,further more to activate Caspase-3,8,9 is the important mechanism of apoptosis induction by allicin.