基于JAK2/STAT3通路研究周氏固金消瘤汤含药血清对人肺腺癌A549细胞侵袭迁移能力的影响  

Effect of drug-containing serum of Zhoushi Gujin Xiaoliu decoction on migration and invasion of human lung adenocarcinoma A549 cells based on JAK2/STAT3 pathway

作　　者：文静 王理槐[1] 侯超 WEN Jing;WANG Lihuai;HOU Chao(The First Hospital of Hunan University of Chinese Medicine,Changsha 410007,China)

机构地区：[1]湖南中医药大学第一附属医院,湖南长沙410007 [2]深圳市中医院,广东深圳518033

出　　处：《陕西中医》2024年第4期451-456,共6页Shaanxi Journal of Traditional Chinese Medicine

基　　金：江苏省自然科学基金资助青年项目(BK20180277);湖南省中医肿瘤临床研究中心项目(2021SK4023);湖南省教育厅科学研究项目(21C0247);广东省深圳市医疗卫生三名工程项目(SZZYSM202211003)。

摘　　要：目的:探讨周氏固金消瘤汤(ZSGJXLT)含药血清对人肺腺癌A549细胞侵袭迁移能力的影响及其机制。方法:应用健康SD大鼠制备ZSGJXLT含药血清与空白血清,将0%(空白血清)及5%、10%、20%、30%、40%ZSGJXLT含药血清作用于A549细胞,CCK-8法测算细胞活性抑制率和半数抑制浓度(IC_(50))。用0%ZSGJXLT(空白对照组)、5%ZSGJXLT(ZSGJXLT低剂量组)、10%ZSGJXLT(ZSGJXLT中剂量组)及20%ZSGJXLT(ZSGJXLT高剂量组)含药血清培养A549细胞,细胞划痕实验及高内涵细胞成像技术观察细胞运动情况;Transwell侵袭及迁移实验观察细胞侵袭及迁移情况;Western blot法检测A549细胞JAK2/STAT3通路相关蛋白表达情况。结果:与空白对照组相比,不同浓度ZSGJXLT含药血清组A549细胞活性受到不同程度的抑制,与时间及浓度呈正相关性,24 h的IC_(50)为25.42%。与空白对照组相比,ZSGJXLT中、高剂量组A549细胞划痕愈合率、细胞移动速度均显著下降(均P<0.05),不同浓度ZSGJXLT含药血清组A549细胞侵袭及迁移数量明显减少(均P<0.05)。与空白对照组比较,ZSGJXLT中、高剂量组A549细胞的p-JAK2、p-STAT3、VEGFA蛋白表达水平显著降低(均P<0.05);不同浓度ZSGJXLT含药血清组MMP-2、MMP-9、N-cadherin蛋白表达有下调趋势,而E-cadherin蛋白表达有上调趋势,呈剂量依赖性,其中ZSGJXLT高剂量组与空白对照组比较差异有统计学意义(均P<0.05)。结论:周氏固金消瘤汤含药血清能抑制A549细胞侵袭迁移能力,其机制可能与调控JAK2/STAT3通路有关。Objective:To investigate the effect of Zhoushi Gujin Xiaoliu decoction(ZSGJXLT) medicated serum on the invasion and migration ability of human lung adenocarcinoma A549 cells and its mechanism.Methods:Healthy SD rats were applied to prepared ZSGJXLT containing serum.The serum containing 0%,5%,10%,20%,30% and 40% ZSGJXLT was used to culture A549 cells.The cell activity inhibition rate and IC_(50)was evaluated by CCK-8 method.The drug-containing serum of 0% ZSGJXLT(blank control group),5% ZSGJXLT(low-dose group),10% ZSGJXLT(middle-dose group) and 20%ZSGJXLT(high-dose group) was used to culture A549 cells.The motility ability of cells was observed by wound healing assay and high-content imaging system analysis.The invasion and migration ability of cells was observed by Transwell invasion and migration assays.The expression of JAK2/STAT3 pathway related proteins were detected via Western blot.Results:Compared with the blank control group,the cell activity of A549 cells were inhibited by ZSGJXLT containing serum obviously in a concentration and time-dependent manner,and IC_(50) of 24 h was 25.42%.The wounding healing rate and movement speed of A549 cells decreased obviously cultured with middle-dose and high-dose ZSGJXLT containing serum(all P<0.05).The invasion and migration ability rate of A549 cells decreased obviously cultured with ZSGJXLT containing serum(all P<0.05).The expression of p-JAK2,p-STAT3,and VEGFA proteins decreased obviously cultured with middle-dose and high-dose ZSGJXLT containing serum(all P<0.05).The expression of MMP-2,MMP-9,and N-cadherin proteins decreased,but the expression of E-cadherin protein of A549 cells increased cultured with ZSGJXLT containing serum in a concentration-dependent manner,in which high-dose ZSGJXLT containing serum group had significant difference(all P<0.05).Conclusion:ZSGJXLT containing serum could inhibit the invasion and migration ability of A549 cells,which might be through inhibiting the activity of JAK2/STAT3 signal pathway.

关 键 词：肺腺癌A549细胞 周氏固金消瘤汤 含药血清 侵袭 迁移 JAK2/STAT3 

分 类 号：R734.2[医药卫生—肿瘤]