基于IL-6/JAK2/STAT3通路探讨斑蝥酸钠对人胃癌细胞增殖、凋亡及炎症免疫的影响  

Effect of sodium norcantharidate on the proliferation,apoptosis and inflammatory immune responses of gastric cancer cells in humans based on IL-6/JAK2/STAT3 pathway

作　　者：王婷 葛尧 张小瑞 WANT Ting;GE Yao;ZHANG Xiao-rui(Shaanxi Energy Institute,Xianyang Shaanxi 712000;School of Basic Medical Sciences,Xi’an Jiaotong University,Xi’an 710061;Department of Pharmacy,The PLA 986th Hospital,Xi’an 710000)

机构地区：[1]陕西能源职业技术学院,陕西咸阳712000 [2]西安交通大学医学部基础医学院,西安710061 [3]中国人民解放军空军第986医院药剂科,西安710000

出　　处：《中南药学》2024年第9期2314-2319,共6页Central South Pharmacy

基　　金：陕西省自然科学基础研究项目青年专项项目(No.2021JQ-887)。

摘　　要：目的探讨去甲斑蝥酸钠(SNCTD)基于白细胞介素-6(IL-6)/酪氨酸蛋白激酶2(JAK2)/信号转导及转录激活因子3(STAT3)通路对胃癌细胞增殖、凋亡和炎症免疫的影响。方法以人胃癌HGC-27细胞为研究对象,加入不同浓度SNCTD(0.25、0.5、1、2、4、-8、16μmol·L^(-1))分别处理24、48h后,用CCK-8法检测各浓度组细胞生长抑制率。不同浓-度SNCTD(1、2、4μmol·L^(-1))作用细胞24h后,ELISA法检测细胞上清液中IL-6水平;流式细胞术检测各浓度组细胞凋亡率;Western blot法和RT-qPCR法分别对IL-6/JAK2/STAT3通路基因(IL-6、GP130、JAK2、STAT3)、细胞增殖和凋亡相关基因(c-Myc、Bcl-2、Bax)进行mRNA和蛋白水平上的表达情况检测。结果与对照组相比,SNCTD组细胞生长抑制率、凋亡率均明显升高(P<0.05);细胞上清液中IL-6水平下降(P<0.01);细胞c-Myc、Bcl-2的mRNA和蛋白表达量均明显下降(P<0.05),而Bax的mRNA和蛋白表达量则明显上升(P<0.05);细胞IL-6、GP130、p-JAK2/JAK2、p-STAT3/STAT3蛋白表达量及JAK2、STAT3 mRNA表达明显下降(P<0.05)。结论SNCTD能有效抑制胃癌细胞增殖并诱导凋亡,其机制可能与SNCTD降低胃癌细胞微环境中炎症因子IL-6的表达有关,通过抑制IL-6/JAK2/STAT3通路的激活,达到调控特定基因的转录与翻译,包括下调c-Myc和Bcl-2、上调Bax的表达。Objective To determine the effect of sodium norcantharidate(SNCTD)on the proliferation,apoptosis and inflammatory immunity of gastric cancer cells,and the role of IL-6/JAK2/STAT3 pathway in these effects.Methods Human gastric cancer HGC-27 cells were used as objects.CCK-8 assay was used to measure the rate of cell proliferation inhibition after the treatment with-various concentrations of SNCTD[0.25,0.5,1,2,4,8,and 16μmol·L^(-1)]for 24 and 48 h.After-the treatment with various concentrations of SNCTD[1,2,and 4μmol·L^(-1)]for 24 h,ELISA was used to measure the concentration levels of IL-6 in the cell supernatant.Flow cytometry was used to measure the apoptosis rate of cells.Western blot and RT-qPCR were used to measure the expressions at mRNA and protein expressions of IL-6/JAK2/STAT3 pathway genes(IL-6,GP130,JAK2,STAT3)and the cell proliferation and apoptosis genes(c-Myc,Bcl-2,Bax)respectively.Results Compared with the control group,the growth inhibition rate and apoptosis rate of the SNCTD groups were significantly increased(P<0.05).The level of IL-6 in the supernatant was decreased(P<0.01).The mRNA and protein expression levels of c-Myc and Bcl-2 were significantly decreased(P<0.05),while Bax mRNA and protein expression increased significantly(P<0.05).The expressions of IL-6,GP130,p-JAK2/JAK2,and p-STAT3/STAT3 protein JAK2 and STAT3 mRNA was decreased(P<0.05).Conclusion SNCTD can effectively inhibit the proliferation and induce the apoptosis of gastric cancer cells,whose mechanism of action may be connected with the fact that SNCTD can reduce the expression of inflammatory factor IL-6 in the tumor microenvironment.The activation of IL-6/JAK2/STAT3 signaling pathway is inhibited to regulate the transcription and translation of specific genes,namely down-regulating the expression of c-Myc and Bcl-2 and up-regulating the expression of Bax.

关 键 词：胃癌 斑蝥酸钠 增殖 凋亡 炎症因子 IL-6/JAK2/STAT3通路 

分 类 号：R285[医药卫生—中药学]