肝细胞生长因子对非小细胞肺癌侵袭和迁移的影响

摘要

Objective To explore that how hepatocyte growth factor ( HGF) effects metastasis on lung cancer and its possible molecular mechanisms .Methods Non-small cell lung cancer ( NSCLC) cells were induced with HGF to established drug-resistance A549/DDP cell line.Real-time PCR and Western blot were applied to detect the expression of HGF on epithelial-mesenchymal transition(EMT)-related markers, including Vimentin and E -cadherin, which in lung cancer cell lines A549 and A549/DDP with different drug -resistance.Cell wound scratch assay and Transwell assay were applied to evaluate the effects of HGF on A 549/DDP and A549 cells′invasion and migration ability before and after the stimulation-induced.Results The expression of HGF in the lung cancer cell A 549/DDP was (2.82 ±0.23) times higher than that in A549 cells (P<0.05).Improving the level of HGF on the cellular microenvironment in A 549/DDP and A549 could stimulate the EMT makers Vimentin enhanced while E -cadherin descend , which the difference was sta-tistically significant (the genetic level P<0.01, protein level P<0.05).The cell wound scratch assay showed that the scratch width of A549/DDP group was significantly higher than that in A549 group (P<0.05).Transwell assay showed that the invasion ability of A549/DDP cells after inducing with HGF was significantly increased than A 549 group ( P<0.01).Conclusion HGF increase invasion and migration ability of A549/DDP and A549 cells via regulationg the ex-pression of EMT-related genes .%目的 探讨肝细胞生长因子(HGF)对非小细胞肺癌(NSCLC)转移的影响及其可能的分子机制.方法 以HGF进行诱导培养NSCLC细胞株A549及其耐药株A549/DDP,应用Real time PCR及Western blot检测诱导前后细胞内上皮间质转化(EMT)相关标志物波形蛋白(vimentin)、钙黏蛋白E(E-cadhe-rin)表达的变化.将细胞种植在细胞培养板,设置未外源性添加HGF的为对照组,外源性添加HGF进行细胞培养的为HGF诱导组,应用细胞划痕以及Transwell实验检测诱导前后两株细胞迁移侵袭能力的变化.结果 HGF在A549/DDP细胞的mRNA表达水平较A549细胞显著升高,差异有统计学意义(P<0.05);提高A549/DDP及A549细胞微环境中HGF水平可诱导细胞vimentin增加、E-cadherin减少,差异均有统计学意义(基因水平P<0.01,蛋白水平P<0.05).细胞划痕实验显示,A549/DDP细胞愈合较A549细胞快,差异有统计学意义(P<0.05),HGF诱导可使细胞侵袭能力增强;Transwell结果显示,HGF诱导组的A549/DDP、A549细胞相对未诱导前细胞的侵袭能力增强,差异均有统计学意义(均P<0.01).结论 HGF促进NSCLC细胞A549/DDP及A549的侵袭迁移,其机制可能与通过调控EMT相关基因的表达相关.