【摘要】 目的:研究组织型转谷氨酰胺酶(tTG)表达与人非小细胞肺癌(NSCLC)细胞株A549和PAα侵袭力的关系,及其与表皮生长因子受体(EGFR)、基质金属蛋白酶-2(MMP-2)等肿瘤侵袭相关基因之间的关系。方法:Transwell法检测细胞株A549和PAα的侵袭力;RT-PCR法、Real-Time PCR法及Western blot检测A549和PAα细胞株中tTG、EGFR和MMP-2的转录和表达水平,观察使用tTG或EGFR抑制剂后PAα细胞中tTG、EGFR、p-EGFR和MMP-2的表达水平变化。结果:PAα细胞株的侵袭能力为A549的165.3倍(P<0.05)。PAα细胞株的tTG、EGFR转录水平为A549细胞株的650.68(P<0.05)和34.4(P<0.001)倍。PAα细胞株的tTG、p-EGFR表达水平高于A549细胞株,MMP-2的转录水平在A549和PAα细胞株之间的差异没有统计学意义(P>0.1),但PAα的MMP-2表达水平高于A549。结论:PAα相对于A549具有高的侵袭力;其机制可能与EGFR的高表达和磷酸化、tTG和MMP-2的高表达有关;tTG可调控MMP-2的水平但它本身不受EGFR信号通路的调控。更多还原

【Abstract】 Objective To explore the relationship between the invasive capacity of human non-small cell lung cancer(NSCLC) cells and level of tissue transglutaminase(tTG) expression or levels of epidermal growth factor receptor(EGFR) and matrix metalloproteinase(MMP-2). Methods Transwell assay was used to detect the invasive capability of A549 cells and PAα cells. Real-time PCR, RT-PCR and Western blotting were used to detect transcription and protein level of tTG, EGFR, and MMP-2 mRNA, and the expression levels of tTG, EGFR, p-EGFR, and MMP-2 after uses of tTG inhibitor or EGFR inhibitor. Results The invasive capacity of PAα was 165.3 times of A549 cells(P < 0.05). The transcription levels of tTG and EGFR mRNA in PAα cells were 650.68 times(P < 0.05) and 34.4 times(P < 0.001) of those in A549 cells. The expression levels of tTG and p-EGFR protein were higher in PAα cells than in A549 cells. There was no significant difference in MMP-2 mRNA transcription level between A549 and PAα cells(P > 0.1), but the expression level of MMP-2 protein was higher in PAα cells than in A549 cells. Conclusions PAα cells has a higher invasive capacity, whose mechanisms are associated with higher expression and phosphorylation of EGFR and higher expressions of tTG and MMP-2. tTG can regulate level of MMP-2 expression, but it is not regulated by the EGFR signaling pathway.更多还原