沉默HIF-1α调控MAPK/ERK信号通路抑制NSCLC转移的分析

Silencing HIF-1αregulates MAPK/ERK pathway and inhibits metastasis of NSCLC

目的探讨HIF-1α在体内及体外对非小细胞肺癌(NSCLC)细胞转移的影响及其作用机制。方法采用实时荧光定量PCR(RT-qPCR)检测HIF-1α在癌旁正常组织、肺鳞癌组织(LUSC)、肺腺癌组织(LUAD)、A549细胞和HEB细胞中的表达量。上调HIF-1α表达后,通过MTT、细胞侵袭和Western blot实验检测过表达HIF-1α对A549细胞增殖、侵袭和EMT的影响。下调HIF-1α表达后,Western blot检测A549细胞ERK和p-ERK蛋白的表达量。THBQ激活MAPK/ERK通路后,分析A549细胞增殖、侵袭和EMT能力。将细胞株植入裸鼠体内,构建移植瘤模型,最后测量裸鼠移植瘤的体积和重量。结果HIF-1α在NSCLC组织的表达水平明显高于癌旁正常组织(P<0.05)。A549细胞中HIF-1α表达水平明显高于HEB细胞(P<0.01),过能够促进A549细胞的增殖和侵袭,N-cadherin蛋白表达量明显上升,E-cadherin蛋白表达量明显下降。下调HIF-1α能够抑制A549细胞内MAPK/ERK通路,且抑制了A549细胞的增殖、侵袭和EMT。下调HIF-1α使裸鼠移植瘤的重量和体积明显减小。结论沉默HIF-1α通过MAPK/ERK信号通路在体内及体外抑制NSCLC转移。

Objective To investigate the effects of HIF-1αon metastasis of non-small cell lung cancer and its mechanism in vitro and in vivo.Methods Quantitative real-time PCR(RT-qPCR)was used to detect the expression of HIF-1αin normal paracancer tissues(Normal),lung squamous cell carcinoma(LUSC),lung adenocarcinoma(LUAD),A549 celland HEB cell.HIF-1αexpression was upregulated,and the effects of overexpression of HIF-1αon the proliferation,invasion,and EMT of A549 cells were detected by MTT,cell invasion,and Western blot.HIF-1αexpression was down-regulated,ERK and p-ERK protein expressions in A549 cells were detected by Western blot.After THBQactivated the MAPK/ERK pathway,the proliferation,invasion and EMT abilities of A549 cells were analyzed.Cell lines were implanted into nude mice to construct a xenograft model.And the volume and weight of the xenograft tumor were measured.Results HIF-1αexpression in non-small cell lung cancer tissues was significantly higher than that in adjacent normal tissues(P<0.05).The expression level of HIF-1αin A549 cells was significantly higher than that in HEB cells(P<0.01).Overexpression of HIF-1αpromoted the proliferation and invasion of A549 cells,and the expression level of N-cadherin protein is significantly increased,while that of E-cadherin is significantly decreased.Down-regulation of HIF-1αinhibited the MAPK/ERK pathway in A549 cells and inhibited the proliferation,invasion and EMT of A549 cells.Downregulation of HIF-1αsignificantly reduced the weight and volume of transplanted tumors in nude mice.Conclusion Silencing HIF-1αinhibited metastasis of non-small cell lung cancer in vivo and in vitro via the MAPK/ERK signaling pathway.