隐丹参酮诱导人食管癌HCE-4细胞凋亡的机制

目的 探讨隐丹参酮对食管癌HCE-4细胞的药效和药理机制。方法 MTT比色法检测隐丹参酮对人食管癌HCE-4和TE-2细胞的杀伤作用。倒置显微镜观察隐丹参酮处理后HCE-4细胞形态学变化。Annexin V-FITC/PI双染法和流式细胞术检测隐丹参酮对HCE-4细胞的诱导凋亡作用、活性氧水平及加入活性氧清除剂NAC后细胞凋亡情况。Western blot检测细胞凋亡相关蛋白的表达量。结果 MTT比色法检测结果显示隐丹参酮对HCE-4和TE-2细胞均具有良好的杀伤作用。经隐丹参酮处理后，HCE-4细胞呈现细胞核固缩、细胞变圆、悬浮细胞增多的现象。Annexin V-FITC/PI双染法和流式细胞术检测结果表明隐丹参酮可以诱导HCE-4细胞发生凋亡，增加细胞内活性氧的水平，预处理NAC后，隐丹参酮的诱导凋亡作用被抑制。Western blot检测结果显示促凋亡蛋白p-JNK、p-p38、Bad、Caspase-3及PARP表达量升高，抗凋亡蛋白p-ERK、p-AKT及Bcl-2表达量降低。结论 隐丹参酮对人食管癌HCE-4细胞具有良好的杀伤作用，其机制可能是隐丹参酮上调ROS的水平，调控MAPK和AKT信号途径，进而诱导食管癌HCE-4细胞发生凋亡。

Apoptotic Effects of Cryptotanshinone on Human Esophageal Carcinoma HCE-4 Cells and Related Mechanism

Objective To explore the effects and pharmacological mechanism of cryptotanshinone (CT) on human esophageal carcinoma HCE-4 cells. Methods MTT assay was performed to determine the cytotoxic effects of CT on HCE-4 and TE-2 cells. The morphological alterations of HCE-4 cells were demonstrated by inverted microscope. Annexin V-FITC/PI double staining and flow cytometry were used to detect the apoptotic effect and reactive oxygen species (ROS) levels of CT in HCE-4 cells. Western blot was performed to investigate the expression levels of apoptotic and upstream signaling pathway-related proteins. Results MTT assay results suggested that CT significantly inhibited the viabilities of esophageal carcinoma HCE-4 and TE-2 cells in a dose-dependent manner. The apoptotic morphology of HCE-4 cells was found under the inverted microscope, such as cell shrinkage, becoming round and the number of suspended cells were increased. Annexin V-FITC/PI double staining and flow cytometry results showed that CT could induce the apoptosis of HCE-4 cells and increase the accumulation of intracellular ROS. Pre-treatment with N-acetyl-L-cysteine (NAC), an ROS scavenger, inhibited the CT-induced apoptosis. Western blot results showed that CT increased the protein expression levels of p-JNK, p-p38, Bad, Caspase-3 and PARP, and also decreased the protein expression levels of p-ERK, p-AKT and Bcl-2. Conclusion Cryptotanshinone inhibits the proliferation and induces the apoptosis of HCE-4 cells via ROS-mediated regulation of MAPK and AKT signaling pathways.