X射线对胃癌细胞和人脐静脉内皮细胞LOX家族成员的不同影响  

Effects of X-ray on LOX family in gastric cancer cells and human umbilical vein endothelial cells

作　　者：王欣怡 牛海亚 邓红 杨新燕 刘一凡 王凯博 徐传皓 韩梅 WANG Xinyi;NIU Haiya;DENG Hong;YANG Xinyan;LIU Yifan;WANG Kaibo;XU Chuanhao;HAN Mei(Department of Pathogen Biology and Medical Immunology,School of Basic Medicine,Ningxia Medical University,Yinchuan 750004,Ningxia,China;Department of Radiology,Changji Branch,the First Affiliated Hospital of Xinjiang Medical University,Changji 831100,Xinjiang,China)

机构地区：[1]宁夏医科大学基础医学院病原生物学与医学免疫学系,银川750004 [2]新疆医科大学第一附属医院昌吉分院放射科,昌吉831100

出　　处：《医学研究与战创伤救治》2024年第1期7-13,共7页Journal of Medical Research & Combat Trauma Care

基　　金：国家自然科学基金(31860255);宁夏大学生创新创业训练计划项目(S202110752011)。

摘　　要：目的比较不同剂量X射线对人低分化粘液样胃腺癌细胞MGC-803和人脐静脉内皮细胞(HUVEC)赖氨酰氧化酶(LOX)家族成员的影响,为了解X射线辐射后不同细胞中基因和蛋白的改变提供依据。方法0、2、4、6、8和10 Gy剂量X射线辐射MGC-803和HUVEC细胞,用细胞增殖实验检测细胞致死率并计算细胞半数致死量(LD 50),确定后序测试剂量组;Realtime PCR、Western blot、ELISA及Amplex Red过氧化氢法分别检测辐射后细胞LOX家族成员mRNA丰度、蛋白相对量、分泌量及LOX酶活性变化;Western blot方法分析X射线辐射对细胞p-p38MAPK和p-Erk1/2MAPK影响。结果X射线辐射后的LD 50胃癌MGC-803为8.2 Gy,HUVEC为6 Gy,后序试验选用0、2、4和6 Gy剂量组。MGC-803细胞经各剂量X射线辐射后LOXL1、LOXL2和LOXL4的mRNA均下调,而LOX的mRNA上调(P<0.05);2 Gy组LOX和LOXL4的蛋白和酶分泌水平低于0 Gy组(P<0.05);4 Gy组LOX、LOXL1、LOXL2和LOXL4的蛋白和分泌低于0 Gy组(P<0.05);6 Gy组LOXL1的蛋白和分泌低于0 Gy组(P<0.05);但LOXL3的mRNA、蛋白和分泌在2 Gy组均高于0 Gy组(P<0.05)。HUVEC中,2 Gy组LOXL1和LOXL3的mRNA、蛋白和酶分泌水平均显著高于0 Gy组(P<0.05);4 Gy组LOX、LOXL1、LOXL3和LOXL4的mRNA、蛋白和酶分泌水平均较0 Gy组升高(P<0.05);6 Gy组LOXL2和LOXL4的mRNA、蛋白和酶分泌水平均较0 Gy组升高(P<0.05)。2 Gy和6 Gy组HUVEC的LOX酶活性增强(P<0.05)。MGC-803细胞的2 Gy和6 Gy组p-p38、p-p42、p-p44较0 Gy组上调(P<0.05);HUVEC经各剂量辐射p-p38、p-p42和p-p44均上调(P<0.05)。结论X射线辐射对两种细胞作用不同,一定剂量X射线辐射上调HUVEC的LOX及其家族成员的产生、分泌和酶活性,下调胃癌细胞MGC-803的除LOXL3外其他成员的产生和分泌。Objective This study is aimed to compare the effects of different doses of X-ray on expressions of lysyl oxidase(LOX)family members in human poorly differentiated mucoid gastric adenocarcinoma cell(MGC-803)and human umbilical vein endothelial cell(HUVEC).Methods MGC-803 and HUVEC were exposed to different doses(0 Gy,2 Gy,4 Gy,6 Gy,8 Gy and 10 Gy)of X-ray.The growth of cells was evaluated using cell proliferation assay,then the Lethal Dose of 50%(LD 50)was calculated to determine the dosage for subsequent research.The abundance of mRNA,relative quantity of protein and levels of secretion of LOX family members in two cell lines were detected through real-time PCR,Western blot(WB)and enzyme-linked immunosorbent(ELISA).Enzyme activity of LOX was tested with Amplex Red hydrogen peroxide assay.The changes of p-p38MAPK and p-ERK1/2 MAPK were analyzed by WB after X-ray exposure.Results The LD 50 of X-ray radiation were 8.2 Gy to MGC-803 and 6 Gy to HUVEC.Compared with 0 Gy,the mRNA abundances of LOXL1,LOXL2 and LOXL4 of gastric cancer cells MGC-803 were down-regulated,but the LOX mRNA was up-regulated after radiation by 2 Gy,4 Gy,6 Gy X-ray(P<0.05).The relative quantity of proteins and levels of enzyme secretion of LOX and LOXL4 were down-regulated after radiation by 2 Gy X-ray(P<0.05).The relative quantity of proteins and levels of enzyme secretion of LOX,LOXL1,LOXL2 and LOXL4 were down-regulated after radiation by 4 Gy X-ray(P<0.05).the relative quantity of proteins and levels of enzyme secretion of LOXL1 were down-regulated after radiation by 6 Gy X-ray(P<0.05).The abundance of mRNA,relative quantity of protein and levels of secretion of LOXL3 were up-regulated under 2 Gy radiation(P<0.05).Compared with 0 Gy,the levels of mRNA,protein and secreting enzyme of LOXL1 and LOXL3 of HUVEC were significantly up-regulated after irradiation by 2 Gy X-ray(P<0.05).The levels of mRNA,protein and secreting enzyme of LOX,LOXL1,LOXL3 and LOXL4 were up-regulated after irradiation by 4 Gy X-ray(P<0.05).The levels of mRNA,protein and s

关 键 词：X射线 赖氨酰氧化酶家族 人脐静脉内皮细胞 胃癌细胞 MAPK信号通路 

分 类 号：R445[医药卫生—影像医学与核医学]