摘要
目的:基于TCGA数据库观察血管生成素样蛋白1(ANGPTL1)在肺癌细胞生物学行为中的作用机制。方法:生物信息数据库TCGA中的肿瘤大数据分析网站GEPIA中查询ANGPTL1在肺癌组织和癌旁正常组织中的表达(癌旁组织52例,肺癌组织503例),同时分析肺癌患者ANGPTL1高表达和低表达与预后的关系。RT-PCR检测ANGPTL1在不同人肺癌细胞和人肺上皮细胞中的表达,检测ANGPTL1在稳转细胞系NCI-H460-ANGPTL1 mimic及阴性对照NCI-H460-NC和空白对照NCI-H460中的表达;MTT及EdU实验检测ANGPTL1对肺癌细胞增殖能力的影响;Transwell检测ANGPTL1对肺癌细胞侵袭能力的影响;划痕实验检测ANGPTL1对肺癌细胞迁移能力的影响;Western blot检测ANGPTL1过表达对肺癌细胞株NCI-H460中Drp1、PGAM5和Bax蛋白表达的影响。结果:TCGA结果显示,肺癌组织中ANGPTL1表达明显低于癌旁正常组织(P<0.05),ANGPTL1高表达肺癌患者生存时间明显长于ANGPTL1低表达患者(P<0.05);MTT实验结果显示,第2、3、4天NCI-H460-ANGPTL1 mimic组细胞增殖率明显低于NCI-H460和NCI-H460-NC组(P<0.05),NCI-H460-ANGPTL1 mimic组48 h EdU阳性率明显低于NCIH460和NCI-H460-NC组(P<0.05);Transwell实验显示,NCI-H460-ANGPTL1 mimic组细胞通过matrigel基质胶的数量明显少于NCI-H460和NCI-H460-NC组(P<0.05);细胞划痕实验结果显示,NCI-H460-ANGPTL1 mimic组细胞迁移率明显低于NCI-H460和NCI-H460-NC组(P<0.05),Western blot检测结果显示,NCI-H460-ANGPTL1 mimic组细胞中PGAM5和Bax蛋白表达明显高于NCI-H460和NCI-H460-NC组,而Drp1表达明显低于NCI-H460和NCI-H460-NC组(P<0.05)。结论:过表达ANGPTL1可能通过抑制Drp1表达激活PGAM5和Bax,抑制肺癌细胞增殖、侵袭和迁移,ANGPTL1是肺癌的新型肿瘤标志物及潜在治疗靶点。
Objective:To observe mechanism of angiopoietin-like protein 1(ANGPTL1)in biological behavior of lung cancer cells based on TCGA database.Methods:Expression of ANGPTL1 in lung cancer tissues and normal tissues adjacent to cancer was inquired from GEPIA,a tumor big data analysis website in biological information database TCGA was queried(including 52 cases adjacent to cancer tissues and 503 cases of lung cancer tissues),and prognostic relationship between high expression and low expression of ANGPTL1 in lung cancer patients was analyzed. RT-PCR was used to detect expression of ANGPTL1 in different human lung cancer cells and human lung epithelial cells,and to detect expression of ANGPTL1 in stable cell line NCI-H460-ANGPTL1 mimic,negative control NCI-H460-NC and blank control NCI-H460. MTT and EdU tests were used to detect effect of ANGPTL1 on proliferation of lung cancer cells. Transwell was used to detect effect of ANGPTL1 on invasion ability of lung cancer cells. Scratch test was used to detect effect of ANGPTL1 on migration ability of lung cancer cells. Effect of ANGPTL1 overexpression on Drp1,PGAM5 and Bax proteins expressions in lung cancer cell line NCI-H460 was detected by Western blot.Results:TCGA results showed that expression of ANGPTL1 in lung cancer tissues was significantly lower than that in normal tissues adjacent to cancer(P<0.05). Survival time of lung cancer patients with high expression of ANGPTL1 was significantly longer than that of patients with low expression of ANGPTL1(P<0.05). MTT assay result showed that cell proliferation rate of NCI-H460-ANGPTL1 mimic group was significantly lower than that of NCI-H460 and NCI-H460-NC group at 2,3 and 4 d(P<0.05). Positive rate of EdU in NCI-H460-ANGPTL1 mimic group at 48 h was obviously lower than that of NCI-H460. Transwell test showed that number of cells passing through matrigel matrix gel in NCI-H460-ANGPTL 1 mic group was significantly lower than that in NCI-H460 and NCI-H460-NC groups(P<0.05). Cell scratch test result showed that cell mobility of
作者
王西勇
丁陈波
戴玉夏
宏林
WANG Xi-Yong;DING Chen-Bo;DAI Yu;XIA Hong-Lin(Suzhou Hospital Affiliated to Anhui Medical University(Suzhou Municipal Hospital),Suzhou 234000,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2021年第11期1366-1370,共5页
Chinese Journal of Immunology
基金
皖北卫生职业学院校级课题(WZK201901)
宿州市科技攻关计划项目(202019)
安徽省高等学校自然科学研究项目(KJ2020A1145)。
关键词
血管生成素样蛋白1
肺癌
增殖
侵袭
迁移
Angiopoietin-like protein 1
Lung cancer
Proliferation
Invasion
Migration
