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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 effect of allicin on the growth of human cervical cancer Hela and Caski cells, the expression of TRAIL and its receptors DR4 and DR5 and the activity of Caspase-3, 8 and 9 before and after the allicin treatment, and to explore the effect of allicin on the apoptosis of cervical cancer cells The role of death and its mechanism. Methods (1) Hela and Caski cells were cultured in vitro from 2009 to 2010 in the Second Hospital of Hebei Medical University. MTT assay was used to detect the inhibitory effect of different concentrations of allicin on the cell growth. (2) The effect of allicin on TRAIL mRNA expression was detected by RT-PCR. (3) Flow cytometry was used to detect the expression of TRAIL and its receptors DR4 and DR5 before and after the allicin treatment. (4) To detect the effect of allicin on the activity of Caspase-3, 8 and 9 in HeLa and Caski cells. Results (1) Allicin had different degrees of growth inhibition on cells. With the increase of drug concentration and the prolongation of action time, cytotoxicity gradually increased. (2) The expression of TRAILmRNA increased significantly after allicin treatment for 12h, the difference between the experimental group and the control group was statistically significant (P <0.05). (3) Allicin treatment of cells after 24h TRAIL protein expression increased. (4) Allicin significantly enhanced the activity of Caspase-3, 8 and 9 after 48h. Conclusion Allicin can inhibit the growth of Hela and Caski cells. Upregulating the expression of TRAIL and activating Caspase-3, 8 and 9 are important mechanisms of allicin-induced apoptosis.