化瘀解毒方含药血清通过JAK2/STAT3通路抑制人肺癌H1299细胞侵袭迁移  

Inhibitory Study of Huayu Jiedu Prescription Medicated Serum on Invasion and Migration of Human Lung Cancer Cell Line H1299 by JAK2/STAT3 Pathway

作　　者：魏征[1,2,3] 徐立然[1] 徐江雁[3] 张爱华[2] 李亚峰[2] 张俊萍[2] WEI Zheng;XU Li-ran;XU Jiang-yan;ZHANG Ai-hua;LI Ya-feng;ZHANG Jun-ping(The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China;The Affiliated Hospital of Henan Academy of Chinese Medicine,Zhengzhou 450004,China;Henan University of Chinese Medicine,Zhengzhou 450046,China)

机构地区：[1]河南中医药大学第一附属医院,郑州450000 [2]河南省中医药研究院附属医院,郑州450004 [3]河南中医药大学,郑州450046

出　　处：《中国实验方剂学杂志》2021年第8期74-80,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：中国博士后基金项目(2018M642760,2019T120626);河南省博士后基金项目(2018-066);河南省中医药科学研究专项(2018ZYZD14);河南省重点研发与推广专项(182102311160);河南省中医管理局拔尖人才项目(豫中医科教[2018]35号);河南省中医管理局青苗人才项目(豫中医科教[2018]16号)。

摘　　要：目的:探讨化瘀解毒方含药血清(HJRMS)对人肺癌细胞(H1299细胞)的增殖、侵袭与迁移的影响及其机制。方法:通过细胞增殖与活性检测(CCK-8)法检测化瘀解毒方含药血清对肺癌细胞增殖的抑制能力;利用划痕实验和Transwell小室法检测肺癌细胞的侵袭与迁移作用;蛋白免疫印迹法(Western blot)检测蛋白酪氨酸激酶2(JAK2),信号传导及转录激活因子3(STAT3),磷酸化JAK2(p-JAK2)和磷酸化STAT3(p-STAT3)蛋白表达水平;实时荧光定量聚合酶链式反应(Real-time PCR)检测JAK2,STAT3 mRNA表达水平。结果:(1)与空白组比较,1%~16%化瘀解毒方含药血清分别处理24,48 h,H1299细胞增殖能力受到显著抑制(P<0.01),并呈一定的浓度依赖性。(2)H1299细胞在经化瘀解毒方含药血清处理24 h,与空白组比较,4%,8%化瘀解毒方含药血清组细胞划痕愈合能力受到抑制(P<0.05,P<0.01)。(3)在侵袭实验和迁移实验中,与空白组比较,2%,4%,8%化瘀解毒方含药血清肺癌细胞透膜数均明显减少(P<0.05,P<0.01)。(4)Western blot显示,与空白组比较,4%,8%化瘀解毒方含药血清组均对肺癌H1299细胞中JAK2/STAT3信号通路相关蛋白(JAK2,p-JAK2,STAT3,p-STAT3)表达具有抑制作用(P<0.05,P<0.01)。(5)与空白组比较,使用8%化瘀解毒方含药血清处理肺癌H1299细胞24 h,JAK2,STAT3mRNA表达水平均下降(P<0.05)。结论:化瘀解毒方含药血清能够抑制肺癌H1299细胞的增殖、侵袭与迁移,其作用机制可能与抑制JAK2/STAT3信号通路有关。Objective:To investigate the effect of Huayu Jiedu prescription medicated serum(HJRMS on the proliferation,invasion and migration of human lung cancer cells(H1299 cells)and its mechanism.Method:Cell counting kit-8(CCK-8)method was used to detect the inhibitory effect of HJRMS on the proliferation of lung cancer cells,the effect of HJRMS on the invasion and migration of H1299 cells were determined by Transwell assay and wound healing assay.The protein expressions of Janus kinase 2(JAK2),signal transduction and activation transcription factor 3(STAT3),phosphorylated JAK2(p-JAK2)and phosphorylated STAT3(p-STAT3)were detected by Western blot,the mRNA expression levels of JAK2 and STAT3 were detected by Real-time quantitative polymerase chain reaction(Real-time PCR).Result:(1)Compared with control group,the proliferation of H1299 cells was significantly inhibited after treatment with1%~16%HJRMS serum for 24,48 h,respectively(P<0.01),and showed a certain concentration dependence.(2)After treatment with HJRMS for 24 h,the scratch healing ability of cells in the 4%,8%HJRMS serum groups was inhibited(P<0.05,P<0.01).(3)Compared with control group,the membrane permeability of H1299 cells in invasion and migration experiments in 2%,4%,8%HJRMS serum groups was decreased significantly(P<0.05,P<0.01).(4)Western blot showed that compared with control group,4%,8%HJRMS serum groups inhibited the expression of JAK2/STAT3 signaling pathway related proteins(JAK2,p-JAK2,STAT3,and p-STAT3)in lung cancer H1299 cells(P<0.05,P<0.01).(5)Compared with control group,the mRNA expression levels of JAK2 and STAT3 in lung cancer H1299 cells treated with 8%HJRMS for 24 h decreased significantly(P<0.05).Conclusion:The HJRMS can inhibit the proliferation,invasion and migration of lung cancer H1299 cells,and its mechanism may be related to the inhibition of JAK2/STAT3 signaling pathway.

关 键 词：化瘀解毒方含药血清 肺癌 侵袭 迁移 

分 类 号：R22[医药卫生—中医基础理论] R242[医药卫生—中医学]