藤梨根提取物对大肠癌LoVo细胞增殖的抑制作用及诱导凋亡的影响

目的:研究藤梨根提取物(ethanol extract from radix of actinidia chinensis,EERAC)对人大肠癌LoVo细胞增殖和凋亡的影响.方法:提取藤梨根抗癌有效活性成分(EERAC),按浓度分为4处理组(10、40、160、320mg/L)和空白对照组(0mg/L).各实验组经作用24、48、72h后,进行一般形态学和AO/EB荧光染色观察;MMT法检测细胞增殖的抑制情况;免疫组织化学(immunohistochemistry,IHC)法测定LoVo细胞中凋亡相关基因Bcl-2、Bax、Caspase-3的蛋白表达变化.结果:与空白对照组比较,一般形态学显示EERAC处理组能使细胞密度减低,增殖变慢;细胞逐渐变大,细胞间接触变松,胞浆中颗粒增多,细胞脱壁现象和周围碎片增多;荧光染色观察可见处理组细胞呈橙红色荧光,细胞核出现碎片状或固缩状的凋亡特征学形态改变,凋亡现象与EERAC的浓度呈正相关性;MTT法检测显示,EERAC处理组对LoVo细胞的最佳作用时间为72h,最大抑制率为79.48%,具有浓度和时间的依赖性(P<0.01);IHC检测结果显示EERAC作用LoVo细胞24h后,Bcl-2表达明显减弱,Bax、Caspase-3表达水平明显增高,Bcl-2/Bax比值下降,差异具有统计学意义(P<0.05),其效应与浓度相关.结论:EERAC具有明显抑制LoVo细胞增殖的作用,其机制可能与降低Bcl-2表达,上调Bax、Caspase-3的表达水平,激活线粒体凋亡途径有关.

Ethanol extract from radix of Actinidia chinensis inhibits cell proliferation and induces apoptosis in human colon carcinoma cell line LoVo

AIM: To explore the regulatory effect of ethanol extract from radix of Actinidia chinensis (EERAC) on cell proliferation and apoptosis in human colon carcinoma cell line LoVo. METHODS: LoVo cells were divided into four treatment groups (10, 40, 160, 320 mg/L EERAC) and blank control group. After treatment for different durations, the morphological alterations of LoVo cells were observed by inverse microscopy and AO/EB fluorescent staining, cell proliferation was detected by MTT assay, and the expression of apoptosis-related proteins Bcl-2, Bax and Caspase-3 was detected by immunohistochemistry. RESULTS: Compared to the control group, EERAC treatment reduced cell density, slowed down cell growth, gradually increased cell size, and resulted in an increase in the number of particles in the cytoplasm and the occurrence of cell detachment. Fluorescence staining showed that treated cells emitted an orange-red fluorescence and showed typical apoptotic features such as nuclear fragmentation and pyknotic nuclei. Apoptosis was positively correlated with EERAC concentrations. MTT assay showed that the opti- mal duration of action was 72 h in EERAC treatment group, and the maximal inhibition rate was 79.48%. The inhibitory effect was concentration- and time-dependent. ICH results showed that the expression of Bcl-2 and the ratio of Bcl-2/Bax were obviously decreased, while the expression levels of Bax and Caspase-3 were significantly increased after treatment for 24 h (P < 0.05). CONCLUSION: EERAC can significantly inhibit LoVo cell proliferation possibly via mechanisms associated with decreasing Bcl-2 expression and increasing Bax and Caspase-3 expression.