摘要
目的探讨体外低氧微环境对胃癌细胞侵袭、迁移及大鼠肉瘤蛋白(Ras)/丝裂原活化蛋白激酶(MAPK)/核因子κB(NF-κB)通路的影响。方法体外培养人胃癌细胞MKN-45,分别在含0、0.5、1.0、1.5 mmol/L氯化钴(CoCl2)培养液中进行培养,CCK-8法检测细胞增殖活力;平板细胞克隆形成实验检测菌落形成;划痕实验检测细胞转移能力;Transwell小室系统检测细胞迁移、侵袭能力;蛋白免疫印迹(Western blot)检测缺氧诱导因子(HIF)-1α蛋白和Ras/MAPK/NF-κB通路有关蛋白的表达。结果随着0、0.5、1.0、1.5 mmol/L CoCl2处理剂量的递增,胃癌MKN-45细胞的增殖抑制率递降(P均<0.01);细胞菌落数量递升(P<0.01);细胞转移抑制率递降(P<0.01),细胞迁移、侵袭数量递升(P均<0.01);其HIF-1ɑ、Ras、MAPK、NF-κB蛋白相对表达量递升(P均<0.01)。上述变化均呈剂量依赖性,不同浓度CoCl2处理组与对照组比较差异均有统计学意义(P均<0.05)。结论低氧微环境能够抑制胃癌细胞的增殖,降低其侵袭、迁移能力,其机制可能与抑制Ras/MAPK/NF-κB通路活性有关。
Objective To investigate the effects of hypoxic microenvironment on invasion and migration of gastric cancer cells and the pathway of rat sarcoma protein(Ras)/mitogen activated protein kinase(MAPK)/nuclear factor κB(NF-κB).Methods Human gastric cancer cells MKN-45 were cultured in vitro and cultured in the medium containing 0,0.5,1.0,1.5 mmol/L cobalt chloride(CoCl2),CCK-8 assay was used to detect cell proliferation;the plate cell clone formation assay was use to detect the colony formation;the scratch test was used to detect the cell transfer ability;Transwell cell system was used to detect cell migration and invasion;Western blot was used to detect the expression of HIF-1α protein and Ras/MAPK/NF-κB pathway related proteins.Results With the increasing dose of CoCl2(0,0.5,1.0,1.5 mmol/L),the proliferation inhibition rate of gastric cancer MKN-45 cells decreased(all P<0.01);the number of cell colonies increased(P<0.01);the cell metastasis inhibition rate decreased(P<0.01),the number of cell migration and invasion increased(all P<0.01),and the relative expression of HIF-1α,Ras,MAPK and NF-κB protein increased(all P<0.01).The above changes were dose-dependent,and the differences between the CoCl2 treatment groups and the control group were statistically significant(all P<0.05).Conclusion Hypoxic microenvironment can inhibit the proliferation and decrease the invasion and migration abilities of gastric cancer cells,its mechanism may be related to inhibit the activity of Ras/MAPK/NF-κB pathway.
作者
于鹏杰
燕速
YU Peng-jie;YAN Su(Department of Gastrointestinal Oncology, Qinghai University Affiliated Hospital, Xining, Qinghai 810000, China)
出处
《中国临床研究》
CAS
2019年第7期907-910,914,共5页
Chinese Journal of Clinical Research
基金
青海省卫生计生指导性科研课题(2015-71)~~
关键词
胃癌
低氧
侵袭
迁移
大鼠肉瘤蛋白
丝裂原活化蛋白激酶
核因子ΚB
缺氧诱导因子-1α
Gastric cancer
Hypoxia
Invasion
Migration
Rat sarcoma protein
Mitogen activated protein kinase
Nuclear factor-κB
Hypoxia inducible factor-1α
