摘要
目的探讨细胞外信号调节激酶(ERK)、血管内皮生长因子(VEGF)和基质金属蛋白酶-9(MMP-9)在4-硝基喹啉1-氧化物(4NQO)诱导的小鼠食管癌细胞中的表达。方法选取雌性BABL/c小鼠200只,分为正常组(50只)和模型组(150只),模型组应用4NQO构建食管癌模型,分为模型10周组、模型15周组和模型20周组,各50只。期间监测体重变化,20周以后均麻醉处死。模型组取食管癌细胞,正常组取正常食管细胞。应用MTT法检测细胞活力,应用RT-PCR及Western blot法检测ERK、VEGF、MMP-9 mRNA和蛋白表达量。结果在造模第2天、第10周、第15周、第20周,正常组小鼠体重比较无统计学差异(P>0. 05),各模型组小鼠体重随造模时间延长显著下降(P<0. 01)。MTT结果显示,在饲养20周后,模型10周组、15周组、20周组食管癌细胞增殖率高于正常食管细胞(P<0. 01);随着造模时间的延长(10周→15周→20周),各模型组食管癌细胞增殖率显著增高,差异有统计学意义(P<0. 01)。RT-PCR法结果显示,随着造模时间延长(10周→15周→20周),模型各组癌细胞内的ERK、VEGF和MMP-9 mRNA相对表达量逐渐增加,且均高于对照组(P<0. 01);Western blot结果显示,随着造模时间延长(10周→15周→20周),模型各组癌细胞内的ERK、VEGF和MMP-9蛋白相对表达量逐渐增加,且均高于对照组(P<0. 01)。结论 ERK、VEGF和MMP-9与食管癌细胞增殖和预后转归直接相关。
Objective To investigate the expression of extracellular signal-regulated kinase(ERK),vascular endothelial growth factor(VEGF) and matrix metalloproteinase-9(MMP-9) in 4-nitroquinoline 1-oxide(4NQO)-induced mouse esophageal cancer cells. Methods A total of 200 female BABL/c mice were randomly divided into 4 groups(normal group,model 10-week group,model 15-week group and model 20-week group;n = 50,each). The 4NQO was used to construct esophageal cancer model in the three model groups. Body weight changes were monitored during the period,and all of them were anesthetized and executed 20 weeks later. The cancerization rate was observed and esophageal cancer cells were taken in the three model groups and esophageal cancer cells were taken from normal group. MTT method was used to detect cell viability,RT-PCR and Western blot were used to detect mRNA and protein expression. Results On the 2nd day,10 th,15th and 20th week,there was no significant difference in the body weight of mice in normal group(P >0. 05),and the weight of mice in three model groups decreased significantly with the time of modeling(P< 0. 01). MTT results showed that after 20 weeks of feeding,the proliferation rate of esophageal cancer cells in the model 10-week group,model 15-week group and model 20-week group was significantly higher than that normal cells(P< 0. 01);with the extension of modeling time(10 weeks → 15 weeks → 20 weeks),the proliferation rate of esophageal cancer cells of each model groups increased significantly(P< 0. 01). The results of RT-PCR showed that expression of ERK、VEGF and MMP-9 mRNA in cancer cells of each model group increased gradually with the prolongation of modeling time(10 weeks→15 weeks→20 weeks)(P< 0. 01). Western blot showed that the expression of ERK,VEGF and MMP-9 protein increased gradually with the prolongation of modeling time(10 weeks→15 weeks→20 weeks)(P< 0. 01). Conclusion ERK、VEGF、MMP-9 may be directly related to the proliferation and prognosis of esophageal cancer cells.
作者
郑银彬
吴国俊
罗晖
汪华
ZHENG Yin-bin;WU Guo-jun;LUO Hui;WANG Hua(Department of Cardiothoracic Surgery,Nanchong Central Hospital,the Second Affiliated Medical College of North Sichuan Medical College,Nanchong,Sichuan 637000,China)
出处
《中国临床研究》
CAS
2020年第2期175-179,共5页
Chinese Journal of Clinical Research
基金
四川省教育厅2018年度科研计划项目(18ZA0207)~~
