ROS/SRC/FAK信号通路在膀胱癌疾病进展中的作用机制研究  

Role of ROS/SRC/FAK signaling pathway in the progression of bladder cancer

作　　者：罗华荣[1] 王天如[1] 陈晨 黄盛松[1] 卞崔冬[1] 孙祖俊 吴登龙[1] Luo Huarong;Wang Tianru;Chen Chen;Huang Shengsong;Bian Cuidong;Sun Zujun;Wu Denglong(Department of Urology Surgery,Tongji Hospital Affiliated to Tongji University,Shanghai 200333,China;不详)

机构地区：[1]同济大学附属同济医院泌尿外科,上海200333 [2]同济大学附属同济医院检验科,上海200333

出　　处：《疑难病杂志》2021年第9期918-923,共6页Chinese Journal of Difficult and Complicated Cases

基　　金：上海市科学技术委员会科研计划项目(18411961100)。

摘　　要：目的观察活性氧(ROS)/类固醇受体共激活因子(SRC)/黏着斑激酶(FAK)信号通路对膀胱癌疾病进展的影响。方法收集2019年6月—2020年12月同济大学附属同济医院泌尿外科经膀胱镜活检、膀胱部分切除的膀胱癌组织标本69份,同时取癌旁组织作为对照,免疫组化检测组织中SRC、FAK蛋白阳性表达,并比较其在不同病理分化程度、浸润程度中的差异。以膀胱癌T24细胞为研究对象,分为对照组(细胞正常培养)、ROS清除剂N,N’-二甲基硫脲(DMTU)干预组(DMTU浓度20μmol/L)、DMTU+SRC激酶抑制剂(PP2)干预组(DMTU浓度20μmol/L,PP2浓度5μmol/L)。ROS检测试剂盒检测细胞中ROS水平;Transwell实验检测细胞迁移、侵袭情况;蛋白免疫印迹(WB)检测细胞中SRC、磷酸化SRC(p-SRC)、FAK、磷酸化FAK(p-FAK)、基质金属蛋白酶9(MMP-9)、钙蛋白酶2(calpain2)蛋白表达情况。结果SRC阳性表达于细胞膜上,FAK阳性表达于细胞浆中,与癌旁组织比较,膀胱癌组织中SRC、FAK阳性比例升高(χ^(2)/P=44.357/0.000、15.019/0.030)。浸润性膀胱癌组织中SRC、FAK表达阳性率高于非浸润性膀胱癌(χ^(2)/P=18.200/0.000、4.681/0.030)。与对照组比较,DMTU干预组ROS相对含量降低(P<0.05),细胞迁移、侵袭数量及细胞中p-SRC/SRC、p-FAK/FAK、MMP-9、calpain2蛋白水平升高(P<0.05);与DMTU干预组比较,DMTU+PP2干预组ROS相对含量升高(P<0.05),细胞迁移、侵袭数量及细胞中SRC、p-SRC/SRC、p-FAK/FAK、MMP-9、calpain2蛋白水平降低(P<0.05)。结论抑制ROS可激活SRC/FAK信号通路,从而增强膀胱癌细胞迁移、侵袭,加重疾病进展。Objective Observe the effect of reactive oxygen species(ROS)/steroid receptor coactivator(SRC)/focal adhesion kinase(FAK)signaling pathway on bladder cancer.Methods From June 019 to December 2020,69 specimens of cancer tissues were biopsied by cystoscopy and partially resected in the Department of Urology,Tongji Hospital Affiliated to Tongji University.At the same time,adjacent tissues were taken as controls.Immunohistochemistry detected positive SRC and FAK proteins in the tissues Expression,and compare its differences in different clinicopathological degrees of differentiation and infiltration.Taking bladder cancer T24 cells as the research object,the cells are divided into control group(cells are cultured normally),ROS scavenger N,N'-dimethylthiourea(DMTU)intervention group(DMTU concentration 20μmol/L intervention),DMTU+SRC kinase inhibitor(PP2)intervention group(co-intervention with DMTU concentration of 20μmol/L and PP2 concentration of 5μmol/L).ROS detection kit detects the level of ROS in cells;Transwell experiment detects cell migration and invasion;Western blotting(WB)detects SRC,phosphorylated SRC(p-SRC),FAK,phosphorylated FAK(p-FAK),Matrix metalloproteinase 9(MMP-9),calpain 2(calpain2)protein expression.Results SRC is located on the cell membrane and FAK is located in the cytoplasm.Compared with the adjacent tissues,the positive ratio of SRC and FAK in bladder cancer tissues is increased(χ^(2)/P=44.357/0.000,15.019/0.030).The positive rate of SRC and FAK expression in bladder cancer tissue was higher in the degree of invasion than that of non-invasion(χ^(2)/P=18.200/0.000,4.681/0.030).Compared with the control group,the relative content of ROS in the DMTU intervention group decreased(P<0.05),cell migration,number of invasions,p-SRC/SRC,p-FAK/FAK,MMP9,and calpain2 protein levels in cells increased(P<0.05);Compared with the DMTU intervention group,the relative content of ROS in the DMTU+PP2 intervention group increased(P<0.05),cell migration,number of invasions,SRC,p-SRC/SRC,p-FAK/FAK,MMP9,calpain2

关 键 词：膀胱癌 活性氧 类固醇受体共激活因子 黏着斑激酶 作用机制 

分 类 号：R737.14[医药卫生—肿瘤]