白花蛇舌草提取物通过Hippo-YAP信号通路抑制胰腺癌SW1990细胞上皮细胞间质转化

Hedyotis diffusa extracts inhibits epithelial-mesenchymal transition of pancreatic cancer SW1990 cells through Hippo-YAP signaling pathway

目的:探讨白花蛇舌草提取物对胰腺癌SW1990细胞上皮细胞间质转化(EMT)的影响,分析其与Hippo-Yap相关蛋白(YAP)信号通路活化的关系。方法:体外培养胰腺癌SW1990细胞,试验分为7组:对照组(不添加任何药物处理),低剂量组(白花蛇舌草提取物20 mg/ml),中剂量组(白花蛇舌草提取物50 mg/ml),高剂量组(白花蛇舌草提取物100 mg/ml),抑制剂组(XMU-MP-1抑制剂),高剂量+抑制剂组,阳性对照组(冬凌草甲素15μmol/L)。采用CCK-8法检测各组细胞增殖情况;流式细胞仪检测SW1990细胞凋亡情况;Western blot检测YAP及其磷酸化蛋白(p-YAP)、E钙黏附蛋白(E-cadherin)、N-钙黏附蛋白(N-cadherin)、波形蛋白(Vimentin)、α平滑肌动蛋白(α-SMA)、转录因子(Snail)表达情况。结果:与对照组相比,白花蛇舌草提取物不同剂量组SW1990细胞增殖抑制率均显著升高(P<0.05),且高剂量组、阳性对照组均显著高于中剂量组、低剂量组(P<0.05);与对照组相比,抑制剂组SW1990细胞凋亡率、p-YAP及E-cadherin蛋白表达量均显著降低(P<0.05),而高剂量组、阳性对照组及高剂量+抑制剂组均显著升高(P<0.05),且高剂量组、阳性对照组显著高于高剂量+抑制剂组(P<0.05);与对照组相比,抑制剂组YAP、N-cadherin、Vimentin、α-SMA、Snail蛋白表达量显著升高(P<0.05),而高剂量组、阳性对照组及高剂量+抑制剂组均显著降低(P<0.05),且高剂量组、阳性对照组显著低于高剂量+抑制剂组(P<0.05)。结论:白花蛇舌草提取物可能通过活化Hippo-YAP信号通路抑制胰腺癌SW1990细胞EMT的发生。

Objective:To investigate the effect of Hedyotis diffusa extracts on epithelial mesenchymal transformation(EMT)of pancreatic cancer SW1990 cells,and to analyze its relationship with activation of Hippo-Yep associated protein(YAP)signaling pathway.Methods:Pancreatic cancer SW1990 cells were cultured in vitro and divided into 7 groups:control group(without any medicine),low-dose group(20 mg/ml of Oldenlandia diffusa extracts),medium-dose group(50 mg/ml of Oldenlandia diffusa extracts),high-dose group(100 mg/ml of Oldenlandia diffusa extracts),inhibitor group(XMU-MP-1 inhibitor),high dose+inhibition group,and positive control group(15μmol/L Ori).Cell proliferation was detected by CCK-8;the apoptosis of SW1990 cells was detected by flow cytometry;the expressions of YAP and its phosphorylated protein(p-YAP),E-cadherin,N-cadherin,Vimentin,alpha smooth muscle actin(α-SMA),and transcription factor(Snail)were detected by Western blot.Results:Compared with the control group,the proliferation inhibition rate of SW1990 cells in different doses of Oldenlandia diffusa extracts was significantly increased(P<0.05),and the high dose group and positive group was significantly higher than the middle dose group and low dose group(P<0.05);compared with the control group,SW1990 cell apoptosis rate,p-YAP and E-cadherin proteins expressions in the inhibitor group were significantly decreased(P<0.05),while the high-dose group,positive group and the high-dose+inhibitor group were significantly increased(P<0.05),the high dose group and positive group was significantly higher than the high dose+inhibitor group(P<0.05);compared with the control group,the expressions of YAP,N-cadherin,Vimentin,α-SMA,and Snail proteins in the inhibitor group increased significantly(P<0.05),while the high-dose group,positive group and the high-dose+inhibitor group were decreased significantly(P<0.05),and the high-dose group and positive group was significantly lower than the high-dose+inhibitor group(P<0.05).Conclusion:Oldenlandia diffusa extracts may inhibit the occurrence of EMT in pancreatic cancer SW1990 cells by activating Hippo-YAP signaling pathway.