摘要
目的:观察输注的异体血小板对人肺癌细胞A549侵袭、转移的影响,初步探讨其作用机制。方法:选取2017年1月至2018年12月于河北医科大学第四医院化疗科就诊输注血小板的89例晚期肺癌患者,实验分为Ctrl组(与培养液共孵育的A549细胞组)、Before组和After组(分别指与输注血小板前和后患者血浆共孵育的A549细胞组)。通过划痕实验和Transwell实验检测与输血小板前后患者血浆共孵育的A549细胞的迁移和侵袭能力,采用Western blotting法检测金属基质蛋白酶(MMPs)及其抑制剂(TIMPs)和上皮-间质转化(EMT)相关蛋白E-cadherin、N-cadherin及Vimentin,以及血管内皮生长因子VEGF及受体2(VEGFR2)的表达水平。结果:After组的A549细胞的划痕愈合率明显高于Before组及Ctrl组([73.67±2.60)%vs(58.33±2.33)%、(35.33±2.03)%,P<0.01或P<0.05],Before组与Ctrl组比较也具有显著差异(P<0.05)。细胞迁移实验结果显示,After组的穿膜细胞数明显高于Ctrl组和Before组([69.67±7.84)vs(18±2.08)、(39.33±2.03)个,均P<0.01]。细胞侵袭实验显示,After组的穿膜细胞数明显高于Ctrl组和Before组([59.34±3.46)vs(18.34±1.56)、(37.58±2.79)个,均P<0.01]。A549细胞与输注血小板前、后的血浆共孵育48 h后,MMP9、MMP2的表达均升高(P<0.05)而其抑制剂TIMP1和TIMP2的水平均下降(P<0.01);EMT相关蛋白N-cadherin、Vimentin表达升高(P<0.05)而E-cadherin表达降低(P<0.01);血管形成相关蛋白VEGF、VEGFR2的表达均升高(P<0.05)。结论:输注异体血小板能促进肺癌A549细胞的侵袭和转移,其作用机制可能与调节EMT、金属基质蛋白酶及血管生长因子相关蛋白的表达有关。
Objective:To observe the effect of allogeneic platelets transfusion on the invasion and metastasis of human lung cancer A549 cells,and to preliminarily explore its mechanism of action.Methods:Eighty-nine patients with advanced lung cancer,who had received platelet transfusion in the Chemotherapy Department of Fourth Hospital of Hebei Medical University between January 2017 and December 2018,were enrolled in this study.The study cells were randomized into Ctrl group(A549 cells co-incubated with culture medium),Before group,and After group(A549 cells co-incubated with plasma Before and After platelet transfusion,respectively).The migration and invasion of A549 cells co-cultured with plasma before and after platelet transfection were detected by Scratch and Transwell experiments.The expression of MMPs,TIMPs and epithelial-mesenchymal transition(EMT)related proteins E-cadherin,N-cadherin and Vimentin,as well as vascular endothelial growth factor(VEGF)and its receptor 2(VEGFR2)were detected by Western blotting(WB)method.Results:The scratch healing ability of A549 cells in After group was significantly higher than that of Ctrl group and Before group[(73.67±2.60)%vs(58.33±2.33)%,(35.33±2.03)%;P<0.01,vs Ctrl group;P<0.05,vs Before group],and there was also a significant difference between Before group and Ctrl group(P<0.05).The results of cell migration experiment showed that the number of transmembrane cells in After group was significantly higher than that in Ctrl group and Before group[(69.67±7.84)vs(18±2.08)and(39.33±2.03),all P<0.01].The cell invasion experiment showed that the number of transmembrane cells in After group was significantly higher than that in Ctrl group and Before group[(59.34±3.46)vs(18.34±1.56)and(37.58±2.79),all P<0.01].When A549 cells were co-incubated with plasma before and after platelet transfusion for 48 h,it was found that the expressions of MMP9 and MMP2 were increased(P<0.05),while their inhibitors TIMP1 and TIMP2 were decreased(P<0.01);the expressions of EMT-related proteins N-cadherin and Vimentin were increased(P<0.05),but E-cadherin was decreased(P<0.01);the expressions of angiogenesis related proteins VEGF and VEGFR2 were increased(P<0.05).Conclusion:Alloplatelets transfusion can promote the invasion and metastasis of lung cancer A549 cells,which may be realized by regulation of the expressions of EMT,metallomatrix protease and vascular growth factor-related proteins.
作者
韩丽娜
赵学涛
马鸣
吴博
赵亮
张璁
单保恩
HAN Lina;ZHAO Xuetao;MA Ming;WU Bo;ZHAO Liang;ZHANG Cong;SHAN Baoen(Department of Blood Transfusion,Fourth Hospital of Hebei Medical University,Shijiazhuang 050011,Hebei,China;Department of Laboratory,Fourth Hospital of Hebei Medical University,Shijiazhuang 050011,Hebei,China;Scientific Research Center,Fourth Hospital of Hebei Medical University,Shijiazhuang 050011,Hebei,China)
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2020年第9期1018-1023,共6页
Chinese Journal of Cancer Biotherapy
基金
河北省医学科学研究重点课题资助项目(No.20170703)。
关键词
血小板
肺癌
A549细胞
侵袭
转移
上皮-间质转化
血管内皮生长因子
金属基质蛋白酶
blood platelet
lung cancer
A549 cell
migrations
invasion
epithelial-mesenchymal transition(EMT)
vascular endothelial growth factor(VEGF)
Matrix metalloproteinase(MMPs)
