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目的:探讨白术内酯Ⅱ对人大肠癌Lovo细胞的抑制作用及其机制。方法:体外培养人大肠癌Lovo细胞,采用不同浓度的白术内酯Ⅱ分别作用于细胞,噻唑蓝(MTT)比色法检测其对细胞生长抑制率,流式细胞术(FCM)检测白术内酯Ⅱ对Lovo细胞凋亡的影响,蛋白免疫印迹法(Western blot)检测白术内酯Ⅱ对大肠癌细胞聚腺苷酸二磷酸核糖转移酶-1(poly ADPribose polymerase1,PARP1),含半胱氨酸的天冬氨酸蛋白水解酶-3(cysteinyl aspartate specific proteinase-3,Caspase-3)蛋白表达水平的影响。结果:与空白组比较,白术内酯Ⅱ质量浓度达到150 mg·L~(-1)时可明显抑制Lovo细胞增殖(P<0.05),随着白术内酯Ⅱ浓度的增高,细胞存活率逐渐下降,当质量浓度超过300 mg·L~(-1)时,细胞存活率降到半数以下;白术内酯Ⅱ(300 mg·L~(-1))可明显促进Lovo细胞的凋亡(P<0.01);白术内酯Ⅱ100 mg·L~(-1)可使PARP1和Caspase-3开始发生剪切,随着药物质量浓度的增加,其剪切作用明显增强。白术内酯Ⅱ可抑制大肠癌Lovo细胞的生长和増殖,并能调节PARP1,Caspase-3蛋白表达,促进细胞凋亡。结论:白术内酯Ⅱ能显著抑制Lovo细胞的增殖并诱导其发生凋亡,其分子机制与促进PARP1的剪切,激活Caspase-3的表达相关。
Objective: To investigate the inhibitory effect of atractyloside Ⅱ on human colorectal cancer Lovo cells and its mechanism. Methods: Human colorectal cancer cell line Lovo was cultured in vitro. Atractyloside II was administered to cells at different concentrations. MTT assay was used to detect cell growth inhibition rate. Flow cytometry (FCM) Ⅱ on the apoptosis of Lovo cells. Western blotting was used to detect the effects of atractylenolide Ⅱ on the expression of poly ADPribose polymerase1 (PARP1), cysteine (Cysteinyl aspartate specific proteinase-3, Caspase-3) protein expression levels. Results: Compared with the blank group, atractylenolide II concentration of 150 mg · L -1 could significantly inhibit the proliferation of Lovo cells (P <0.05). With the increase of atractylenolide Ⅱ concentration, the cell viability gradually increased (P <0.05), and the cell viability decreased to less than half when the concentration was over 300 mg · L -1; atractylenolide Ⅱ (300 mg · L -1) significantly increased the apoptosis of Lovo cells (P <0.01). Atractylenolide Ⅱ 100 mg · L -1 could induce the cleavage of PARP1 and Caspase-3. With the increase of drug concentration, the cleavage of PARP1 and Caspase-3 increased significantly. Atractyloside II can inhibit the growth and proliferation of colorectal cancer Lovo cells, and regulate PARP1, Caspase-3 protein expression and promote apoptosis. CONCLUSION: Atractylenolide Ⅱ can significantly inhibit the proliferation and induce the apoptosis of Lovo cells. The molecular mechanism is related to the cleavage of PARP1 and the activation of Caspase-3 expression.