蜈蚣提取物对人肝癌HepG2细胞STAT3信号通路的影响  

Effect of Scolopendra subspinipes extracts on STAT3 signaling pathway of human hepatocellular carcinoma HepG2 cells

作　　者：廖柳[1] 刘晓斌[1] 周青[2] 王志琪[3] 田莎[4] 陈园[1] 田雪飞[4] LIAO Liu LIU Xiao-bin ZHOU Qing WANG Zhi-qi TIAN Sh CHEN Yuan TIAN Xue-fei(Post-Graduate School, Hunan University of Chinese Medicine, Changsha 410208, China The First Affiliate Hospital, Hunan University of Chinese Medicine, Changsha 410007, China College of Pharmacy, Hunan University of Chinese Medicine, Changsha 410208, China College of Intergrated Chinese and Western Medicine, Hunan University of Chinese Medicine, Changsha 410208, China)

机构地区：[1]湖南中医药大学研究生学院,湖南长沙410208 [2]湖南中医药大学第一附属医院,湖南长沙410007 [3]湖南中医药大学药学院,湖南长沙410208 [4]湖南中医药大学中西医结合学院,湖南长沙410208

出　　处：《中草药》2017年第5期930-934,共5页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(81473617);湖南省高层次卫生人才"225工程"项目资助(湘卫人发2013-13号)

摘　　要：目的观察蜈蚣提取物作用于肝癌Hep G2细胞后对信号传导与转录激活因子3(STAT3)信号通路与磷酸化相关蛋白的表达以及细胞转移侵袭能力的影响,探讨蜈蚣提取物介导STAT3信号通路抗肝癌侵袭转移的调控作用机制。方法梯度质量浓度(300、600、1 200、2 400μg/m L)的蜈蚣提取物处理人肝癌Hep G2细胞,采用CCK8法检测细胞增殖抑制率,计算半数抑制浓度(IC50)。将人肝癌细胞分为对照组,蜈蚣提取物250、500μg/m L组,5-氟尿嘧啶(5-FU)对照组,蜈蚣提取物处理人肝癌细胞48 h时,Transwell细胞侵袭实验检测Hep G2细胞的侵袭能力变化,Western blotting方法检测STAT3、p-STAT3、血管内皮生长因子(VEGF)、基质金属蛋白酶-2(MMP-2)蛋白的表达情况。结果 CCK8方法检测显示,蜈蚣提取物对人肝癌Hep G2细胞增殖有明显抑制作用(P<0.05),呈剂量依赖性,IC50(48 h)值为508.3μg/m L;Transwell细胞侵袭实验结果显示,蜈蚣提取物处理48 h时,人肝癌Hep G2细胞的侵袭能力明显降低,蜈蚣提取物250、500μg/m L组和5-FU组透膜细胞数显著少于对照组(P<0.05);与250μg/m L蜈蚣提取物组比较,500μg/m L蜈蚣提取物组、5-FU组透膜细胞数更少(P<0.05)。Western blotting实验结果显示,蜈蚣提取物处理48 h时,STAT3通路主要以p-STAT3表达下调为主,250、500μg/m L蜈蚣提取物组p-STAT3均较对照组下调(P<0.05、0.01);500μg/m L蜈蚣提取物组处理后的MMP-2、VEGF蛋白表达下调,与对照组比较差异显著(P<0.05),与250μg/m L蜈蚣提取物组比较,MMP-2表达下调更显著(P<0.05)。结论蜈蚣提取物主要通过降低STAT3磷酸化调控STAT3相关信号通路的过度活化,从而降低下游靶蛋白MMP-2、VEGF的表达,抑制人肝癌细胞的增殖及转移侵袭能力。Objective To observe the effect of Scolopendra subspinipes extracts（SSE） on signal transducer and activator of transcription 3（STAT3） signaling pathway and phosphorylation of its protein expression as well as the regulation mechanisms of HepG2 cells proliferation, invasion, and metastasis after SSE exposure. Methods HepG2 cells were processed with SSE of gradient concentration（300, 600, 1 200, and 2 400 μg/m L）. The inhibitory effect of SSE on HepG2 cell proliferation was evaluated by CCK8 method. Subsequent experimental concentration was set from IC50 result of CCK8 methods. HepG2 cells were divided into control, SSE（250 and 500 μg/m L）, and 5-FU groups. After HepG2 cells were treated with SSE for 48 h, Transwell Chambers detected the invasion of HepG2 cells and Western blotting demonstrated expression and activation of STAT3, p-STAT3 and VEGF, MMP-2 protein. Results CCK8 method showed that SSE had obvious inhibition effect on human HepG2 cells proliferation with dose dependent effect（P 〈 0.05）. The IC50 of SSE was 508.3 μg/m L at 48 h. Transwell result showed invasive ability of human HepG2 cells was significantly reduced compared with control group after SSE worked to cells for 48 h（P 〈 0.05）. Compared with 250 μg/m L SSE group, the number of membrane cells in 500 μg/m L SSE and 5-FU groups were less（P 〈 0.05）. Western blotting analysis showed that STAT3 signaling pathway was mainly down regulated by p-STAT3 expression after SSE worked to cells for 48 h. Compared with control group, the p-STAT3 expression of SSE was down-regulated（P 〈 0.05）. The MMP-2 and VEGF protein expression of 500 μg/m L SSE decreased compared with control group（P 〈 0.05）. The MMP-2 protein expression of 500 μg/m L SSE group had obvious difference compared with the 250 μg/m L SSE group（P 〈 0.05）. Conclusion SSE regulate the activation of human HepG2 cells STAT3 signaling pathway by STAT3 phosphorylation down-regulation and reduce the expression of MMP-2 and VEGF downstream target

关 键 词：蜈蚣 肝癌 信号传导与转录激活因子3信号通路(STAT3) HEPG2 血管内皮生长因子 基质金属蛋白酶-2 

分 类 号：R285.5[医药卫生—中药学]