大黄酸对非小细胞肺癌A549细胞增殖、迁移和侵袭能力的影响及其机制

目的：探讨大黄酸对非小细胞肺癌（NSCLC）A549细胞增殖、迁移和侵袭能力的影响，并阐明其机制。方法：将NSCLC A549细胞分为对照组和低、中及高剂量大黄酸组，采用不含大黄酸的培养基和含有质量浓度为5、10和20 μmol·L^-1大黄酸的培养基分别培养细胞24、48和72 h。采用MTT法检测各组A549细胞增殖率，流式细胞术检测A549细胞凋亡率以及不同细胞周期A549细胞百分比，Western blotting法检测各组A549细胞中半胱氨酸蛋白酶3（Caspase-3）、半胱氨酸蛋白酶9（Caspase-9）、细胞色素C（Cyt-C）和凋亡诱导因子（AIF）蛋白表达水平，划痕实验和Transwell实验检测各组A549细胞迁移和侵袭能力。结果：大黄酸处理24h后，与对照组比较，低、中和高剂量大黄酸组A549细胞增殖率明显降低（P<0.05），细胞迁移距离明显减少，侵袭细胞数量明显减少；中和高剂量大黄酸组A549细胞凋亡率明显升高（P<0.05）；高剂量大黄酸组G₁期细胞百分比明显降低（P<0.05）。大黄酸处理48 h后，与对照组比较，中和高剂量大黄酸组A549细胞中Caspase-3、Caspase-9、Cyt-C和AIF蛋白表达水平明显升高（P<0.05）。结论：大黄酸可抑制A549细胞的增殖、迁移和侵袭能力，其作用机制可能与上调Caspase-3、Caspase-9、Cyt-C和AIF蛋白表达有关。

Effects of rhein on proliferation,migration and invasion abilities of non-small cell lung cancer A549 cells and their mechanisms

Objective: To investigate the effects of rhein on the proliferation, migration and invasion of the non-small cell lung cancer (NSCLC) A549 cells, and to elucidate the mechanisms. Methods: The NSCLC A549 cells were divided into control group and low, medium and high doses of rhein groups. The cells in each group were cultured for 24, 48 and 72 h in the culture medium without rheic and the culture medium containing 5, 10 and 20 μmol·L^-1 rhein. MTT method was used to analyze the proliferation rates of A549 cells in various groups. Flow cytometry was used to detect the apoptotic rates and the percentages of A549 cells at different cell cycles. Western blotting method was used to detect the expression levels of cysteine protease -3(Caspase-3), cysteine protease -9(Caspase-9), cytochrome C(Cyt-C) and apoptosis-induced factor (AIF) protines in the A549 cells in various groups. Scratch test and Transwell assay were used to detect the abilities of migration and invasion of A549 cells. Results: After treated with rhein for 24 h, compared with control group, the proliferation rates of the A549 cells in low, medium and high doeses of rhein groups were significantly decreased(P<0.05), the migration distances of cells were significantly reduced, and the number of invasion cells was significantly reduced; the apoptotic rates of the A519 cells in medium and high doses of rhein groups were significantly increased(P<0.05);the percentage of A519 cells in G₁ phase in high dose of rhein group was significantly reduced(P<0.05). After treated with rhein for 48 h, compared with control group, the expression levels of Caspase-3, Caspase-9, Cyt-C and AIF proteins in medium and high doses of rhein groups were significantly increased (P<0.05). Conclusion: Rhein can inhibit the proliferation, migration and invasion abilities of A549 cells and its mechanism may be related to increasing the expressions of Caspase-3, Caspase-9, Cyt-C and AIF proteins.