一种新型查耳酮衍生物C13通过ErbB4/PI3K/AKT信号通路抑制人胃癌细胞的生长

A novel chalcone derivative C13 inhibits the growth of human gastric cancer cells through suppressing ErbB4/PI3K/AKT signaling pathway

3ʹ-羟基-4ʹ-甲氧基-2-羟基-5-溴查耳酮(以下简称C13)是本课题组对龙血竭抗肿瘤活性化合物DHMMF进行结构修饰过程中获得的一个新型查耳酮衍生物。本研究探讨了C13对人胃癌HGC-27、AGS细胞增殖、凋亡的影响及其潜在的作用机制。首先采用噻唑蓝(methyl thiazolyl tetrazolium,MTT)、集落形成实验、5-乙炔基-2′-脱氧尿苷(5-ethynyl-2ʹ-deoxyuridine,EdU)染色法发现C13能显著抑制人胃癌HGC-27、AGS细胞的增殖能力。采用流式细胞术和免疫印迹法检测发现C13能诱导人胃癌HGC-27、AGS细胞发生凋亡,并上调剪切的聚腺苷酸二磷酸核糖转移酶(cleaved-poly ADP-ribose polymerase,cleaved-PARP)的蛋白水平。转录组测序分析结果显示C13可能通过调控人表皮生长因子受体4/磷脂酰肌醇3-激酶/蛋白激酶B(Erb-B2 receptor tyrosine kinase 4/phosphoinositide 3-kinase/AKT,ErbB4/PI3K/AKT)信号通路发挥抗胃癌作用。最后免疫印迹实验结果表明,C13处理后能下调人胃癌细胞中ErbB4总蛋白及磷酸化水平,并能够抑制其下游PI3K及AKT蛋白磷酸化水平,进一步验证了转录组测序结果。综上所述,新型查耳酮衍生物C13能够显著抑制人胃癌HGC-27、AGS细胞增殖并诱导其发生凋亡,其机制可能与下调ErbB4/PI3K/AKT信号通路有关。本研究能够为胃癌治疗提供一个候选研究药物。

3ʹ-Hydroxy-4ʹ-methoxy-2-hydroxy-5-bromochalcone(hereinafter referred to as C13)is a novel chalcone derivative obtained in the process of structural modification of DHMMF,the antitumor active compound of Resina Draconis,in our laboratory.In this study,we investigated the effects of C13 on the proliferation and apoptosis of human gastric cancer HGC-27 and AGS cells and its potential mechanism of action.Firstly,through methyl thiazolyl tetrazolium(MTT),colony formation assay,and 5-ethynyl-2'-deoxyuridine(EdU)staining,we found that C13 inhibited the proliferation ability of human gastric cancer HGC-27 and AGS cells.Using flow cytometry and Western blot,it was found that C13 induced apoptosis in human gastric cancer HGC-27 and AGS cells,and up-regulated the protein level of cleaved poly ADP-ribose polymerase(cleaved-PARP).The results of RNA sequencing analysis showed that the Erb-b2 receptor tyrosine kinase 4/phosphoinositide 3-kinases/AKT(ErbB4/PI3K/AKT)signaling pathway may be involved in anti-gastric cancer activity of C13.Finally,the results of immunoblotting assay showed that C13 treatment down-regulated the protein levels of ErbB4 and phospho-ErbB4,as well as down-regulated the phosphorylation levels of PI3K and AKT in human gastric cancer HGC-27 and AGS cells,which verified the results from RNA-seq analysis.In conclusion,C13 inhibited the proliferation and induced apoptosis of human gastric cancer cells,which may be related to the down-regulation of ErbB4/PI3K/AKT signaling pathway.This study may provide a candidate drug for the treatment of gastric cancer.