摘要
目的探讨芦荟大黄素对大肠癌LOVO细胞自噬相关通路Akt/mTOR、凋亡及迁移的影响。方法将大肠癌LOVO细胞分为对照组（正常培养）、低剂量组（LOVO细胞＋10 μmol/L芦荟大黄素培养30 min）、中剂量组（LOVO细胞＋30 μmol/L芦荟大黄素培养30 min）和高剂量组（LOVO细胞＋50 μmol/L芦荟大黄素培养30 min）。采用CCk-8法检测各组细胞培养24、48、72 h细胞增殖抑制率；流式细胞仪检测各组细胞凋亡率；采用Transwell小室检测各组细胞迁移能力；蛋白质印迹法（Western blot）检测各组细胞IC3-Ⅰ、IC3-Ⅱ、Beclin-1、mTOR、p-mTOR、Akt及p-Akt的表达。结果低剂量组LOVO细胞增殖抑制率、迁移能力、凋亡率与对照组比较差异无统计学意义（P〉0.05），中、高剂量组LOVO细胞增殖抑制率、迁移能力降低，凋亡率增加，与对照组比较差异有统计学意义（P〈0.05），并呈剂量依赖性（P〈0.05）。低剂量组细胞检测IC3-Ⅰ、IC3-Ⅱ、Beclin-1、mTOR、p-mTOR、Akt及p-Akt表达与对照组比较差异无统计学意义（P〉0.05），中、高剂量组IC3-Ⅰ、IC3-Ⅱ、Beclin-1表达增加（0.43 ± 0.03、0.59 ± 0.04比0.16 ± 0.00，0.57 ± 0.07、1.06 ± 0.17比0.34 ± 0.02，0.37 ± 0.02、0.49 ± 0.01比0.13 ± 0.00），mTOR、p-mTOR、Akt及p-Akt表达减少（0.85 ± 0.08、0.37 ± 0.02比2.08 ± 0.07，1.42 ± 0.09、1.19 ± 0.02比1.97 ± 0.11，0.97 ± 0.00、0.84 ± 0.06比1.19 ± 0.02，0.43 ± 0.02、0.31 ± 0.01比0.98 ± 0.08），与对照组比较差异有统计学意义（P〈0.05），并呈剂量依赖性（P〈0.05）。结论芦荟大黄素可能通过降低Akt/mTOR信号通路表达及其磷酸化水平，达到诱导大肠癌LOVO细胞自噬、凋亡，并抑制其生长、迁移能力的作用。
ObjectiveTo investigate the effect of aloe-emodin on Akt/mTOR, apoptosis and migration of autophagy-related pathway in colorectal cancer LOVO cells.MethodsThe LOVO cells were divided into control group （normal culture）, low dose group （LOVO cells ＋ 10 μmol/L aloe-emodin culture 30 min）, medium dose group （LOVO cells ＋ 30 μmol/L aloe-emodin culture 30 min） and high dose group （LOVO cells ＋ 50 μmol/L aloe veratin culture 30 min）. The cell proliferation rate was measured by CCk-8 method at 24, 48 and 72 h respectively. The apoptotic rate of each group was detected by flow cytometry. The migration ability of each group was detected by Transwell chamber. Western blot was used to detect the expression of IC3-Ⅰ, IC3-Ⅱ, Beclin-1, mTOR, p-mTOR, Akt and p-Akt.ResultsCompared with that of the control group, the cell rate, migration ability and apoptosis rate of the low dose group was not significantly changed （P 〉 0.05）. The cell rate and migration ability of the medium and high dose groups was significantly decreased and the apoptosis rate was significantly increased：0.43 ± 0.03, 0.59 ± 0.04 vs. 0.16 ± 0.00; 0.57 ± 0.07, 1.06 ± 0.17 vs. 0.34 ± 0.02; 0.37 ± 0.02, 0.49 ± 0.01 vs. 0.13 ± 0.00, P 〈 0.05, there was dose dependent （P 〈 0.05）. Compared with those of the control group, the expressions of IC3-Ⅰ, IC3-Ⅱ, Beclin-1, mTOR, p-mTOR, Akt and p-Akt in the low dose group showed no significant change （P 〉 0.05）, those were decreased in middle and high dose groups which showed significant differences： 0.85 ± 0.08, 0.37 ± 0.02 vs. 2.08 ± 0.07; 1.42 ± 0.09, 1.19 ± 0.02 vs. 1.97 ± 0.11; 0.97 ± 0.00, 0.84 ± 0.06 vs. 1.19 ± 0.02; 0.43 ± 0.02, 0.31 ± 0.01 vs. 0.98 ± 0.08, P 〈 0.05, there were dose dependent （P 〈 0.05）.ConclusionsAloe-emodin can promote autophagy, apoptosis there inhibit the growth, migration of colorectal cancer LOVO cells by reducing the expression of Akt/mTOR signaling pathway and reducing its phosphorylation level.
作者
郑建军
蔡沣
刘开渊
Zheng Jianjun;Cai Feng;Liu Kaiyuan.(Department of Gastroenterology, 95 Military Hospital of China, Fujian Putian 351100, China)
出处
《中国医师进修杂志》
2018年第2期144-148,共5页
Chinese Journal of Postgraduates of Medicine
关键词
芦荟大黄素
结直肠肿瘤
自噬
增殖
迁移
Aloe-emodin
Colorectal neoplasm
Autophagy
Proliferation
Migration
