论文部分内容阅读
目的:探讨南蛇藤总萜(COE)是否通过诱导自噬来抑制结直肠癌细胞增殖。方法:用不同浓度的COE处理体外培养的大肠癌HT-29细胞,噻唑蓝(MTT)法检查细胞存活率,透射电镜(TEM)观察细胞形态,免疫印迹法(Western blot)检测COE对细胞自噬相关蛋白表达的影响,并采用自噬抑制剂3-甲基腺嘌呤(3-MA)联合COE干预HT-29细胞,用吖啶橙染色细胞、MTT法检测3-MA对COE抑制HT-29细胞增殖的影响。结果:COE抑制HT-29细胞的增殖,作用24h的IC50为156μg/m L;随着COE浓度的升高,TEM示细胞中自噬体明显增多,和COE治疗组比较,COE联合3-MA组的细胞生存率升高,而自噬泡(AVO)的数量显著下降,微管相关蛋白-Ⅱ(LC3-Ⅱ)表达也降低。结论:COE可能通过上调Bcl-2同源结构域蛋白抗体-1(Beclin-1)和LC3-Ⅱ的表达来增加HT-29细胞的自噬活性,从而抑制HT-29细胞的增殖作用。
Objective: To investigate whether COE can inhibit the proliferation of colorectal cancer cells by inducing autophagy. Methods: HT-29 cells were cultured with different concentrations of COE. Cell viability was examined by MTT assay. Cell morphology was observed by transmission electron microscopy (TEM) MAH and COE were used to intervene HT-29 cells. Acridine orange was used to stain the cells. MTT assay was used to detect the effect of 3-MA on HT-29 cells induced by COE. HT-29 cells were treated with 3-methyladenine 29 cell proliferation. RESULTS: COE inhibited the proliferation of HT-29 cells with an IC50 of 156 μg / m L for 24 h. With the increase of COE concentration, TEM showed a marked increase of autophagosomes. Compared with COE group, COE combined with 3-MA Group increased the cell survival rate, whereas the number of autophagic vacuoles (AVO) significantly decreased and the expression of microtubule-associated protein-Ⅱ (LC3-Ⅱ) also decreased. CONCLUSION: COE may increase the autophagy activity of HT-29 cells by up-regulating the expression of Beclin-1 and LC3-Ⅱ, thereby inhibiting the proliferation of HT-29 cells.