摘要
目的研究顺铂（DDP）抑制He La细胞增殖的分子机制。方法用顺铂0.02~75μmol·L-1处理He La细胞24或48 h,CCK-8法检测细胞存活,划痕实验检测细胞迁移力,流式细胞仪检测细胞周期,实时荧光定量PCR（q-PCR）检测转移抑制基因1（MTSS1）基因的表达,Western蛋白质印迹法检测磷酸化细胞外信号调节激酶（p-ERK）、磷酸化丝氨酸/苏氨酸激酶（p-AKT）和转移抑制基因1（MTSS1）蛋白水平。结果不同浓度DDP处理He La细胞24或48 h,可明显抑制He La细胞增殖（P〈0.05）,细胞的半数抑制浓度（IC50）值分别为4.14和11.82μmol·L-1;DDP处理组He La细胞较对照组迁移能力下降（P〈0.05）;DDP使He La细胞周期阻滞于S期;DDP抑制He La细胞MTSS1的基因表达,存在明显的浓度效应关系（r24 h=-0.965,P〈0.01;r48 h=-0.953,P〈0.01）;p-ERK,p-AKT及胞内MTSS1蛋白表达水平均随DDP浓度增加显著下降（pERK：r24 h=-0.875,P〈0.01;r48 h=-0.966,P〈0.01;p-AKT：r24 h=-0.831,P〈0.01;r48 h=-0.863,P〈0.01;MTSS1：r24 h=-0.969,P〈0.01;r48 h=-0.988,P〈0.01）。结论 DDP可能通过下调MTSS1的表达水平并抑制ERK和AKT信号通路的激活而抑制宫颈癌He La细胞增殖和迁移。
OBJECTIVE To study the molecular mechanism of cisplatin（DDP）by which He La cell growth and proliferation are inhibited. METHODS Cultured He La cells were treated with DDP 0.02-75 μmol · L- 1for 24 or 48 h. CCK- 8 assay was used to determine the cell proliferation. The wound scratch assay was used to detect the cell migration and invasion. Flow cytometry was used to detect the cell cycle arresting. q- PCR was used to test the expression of metastasis suppressor gene 1（MTSS1）m RNA. Western blot was used to determine protein levels of MTSS1,phosphorylated-extracel ular signal-regulated kinase（p-ERK）and phosphorylated-serine-threonine kinase（p-AKT）. RESULTS Following the treatment with DDP for 24 or 48 h,the proliferation of He La cells was inhibited significantly（P〈0.05）,the value of the half inhibitory concentration（IC50）of cells was 4.14 and 11.82 μmol · L- 1.Migration and invasion activity of He La cells were reduced according to the wound scratch assay（P〈0.05）. Flow cytometry results showed that the cel cycle was arrested at S phase. q-PCR results showed that MTSS1 m RNA expression changed with DDP in a concentration-dependent manner（r24 h=-0.965,P〈0.01;r48 h=-0.953,P〈0.01）. Western blot showed that the protein levels of MTSS1,p-ERK and p-AKT expression declined significantly with the increase in DDP concentrations（p-ERK：r24 h=-0.875,P0.01;r48 h=-0.966,P〈0.01. p-AKT：r24 h=-0.831,P〈0.01;r48 h=-0.863,P〈0.01. MTSS1：r24 h=-0.969,P〈0.01;r48 h=-0.988,P〈0.01）. CONCLUSION DDP treatment inhibits He La growth and proliferation by interfering with the MTSS1 expression and disturbing the activation of ERK and AKT signaling pathways.
出处
《中国药理学与毒理学杂志》
CAS
CSCD
北大核心
2016年第4期350-355,共6页
Chinese Journal of Pharmacology and Toxicology
基金
国家自然科学基金(31070996)
空军总医院科研基金课题(kz2013011)~~
