摘要
目的通过MTT细胞活性实验、Western蛋白印记实验,证明E804能够对MGC803等胃癌细胞系产生抑制细胞活性的作用,提高自噬标记物的表达,促进MGC803细胞进行自噬过程,并且探究E804的药理机制.方法正常培养的MGC-803与MKN-45细胞作为阴性对照组,实验组将E804按照相应梯度浓度处理24 h,部分实验加入白介素-6浓度100 ng/mL处理2 h作为阳性对照组.采用MTT实验、蛋白印记实验和裸鼠成瘤模型建立实验来分析两组细胞的细胞活性、细胞的自噬标记物表达升高情况及检测检测移植瘤直径变化.结果随着E804浓度的上升,MGC-803与MKN-45两组细胞的细胞活性均降低,差异有统计学意义(P<0.05),E804处理后到自噬标记物LC3-B及Beclin-1的表达量显著上升,并且呈现明显的剂量依赖效应;比较对照组载瘤鼠与给药组载瘤鼠肿瘤最大径随时间的变化曲线,给药组的肿瘤生长速度显著慢于对照组,有统计学差异(P<0.05).结论 E804通过抑制S t a t3活化促进胃癌细胞自噬活动来抑制胃癌细胞生长,为胃癌的联合治疗提供了新的思路.
AIM To explore the pharmacological mechanism of E804,a derivative of indirubin, by investigating whether it can inhibit the viability of gastric cancer cells, alter the expression of markers of autophagy, and promote the autophagy activity.METHODS MGC-803 and MKN-45 cells were treated with different concentrations of E804. Non-treated MGC-803 and MKN-45 cells were used as negative controls, and those treated with interleukin-6（100 ng/mL） for 2 h were used as a positive control. After treatment, cell viability was detected by MTT assay. The expression of autophagy markers was detected by Western blot. The diameter of transplantable tumor in a subcutaneous xenotransplanted tumor model was also measured.RESULTS The viability of both MGC-803 and MKN-45 cells declined significantly after treatment（P〈0.05）. E804 increased the expression of LC3-B and Beclin-1, two markers of autophagy, in a dose-dependent manner. When comparing the maximal diameters of tumor in the control group（non-treated） and experimental group（treated with E804）, the speed of tumor growth in the experimental group was significantly lower than that of the control group（P〈0.05）.CONCLUSION E804 can promote autophagy of gastric cancer cells to suppress their growth, at least partly by inhibiting the activation of Stat3.
出处
《世界华人消化杂志》
CAS
2017年第36期3184-3190,共7页
World Chinese Journal of Digestology
