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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。
Objective: To study the relationship between the expression of tissue transglutaminase (tTG) and the invasiveness of human non-small cell lung cancer (NSCLC) cell lines A549 and PAα, and the relationship with the expression of epidermal growth factor receptor (EGFR), matrix metalloproteinase-2 (MMP-2) and other tumor invasion-related genes. Methods: Transwell assay was used to detect the invasiveness of cell lines A549 and PAα. The transcription and expression levels of tTG, EGFR and MMP-2 in A549 and PAα cell lines were detected by RT-PCR, Real-Time PCR and Western blot. Or EGFR inhibitor PAα cells after tTG, EGFR, p-EGFR and MMP-2 expression levels change. Results: The invasion ability of PAα cells was 165.3 times that of A549 (P <0.05). The tTG and EGFR transcriptional levels of PAα cells were 650.68 (P <0.05) and 34.4 (P <0.001) fold higher than that of A549 cells. The expression level of tTG and p-EGFR in PAα cell line was higher than that in A549 cell line. There was no significant difference in the transcription level of MMP-2 between A549 and PAα cell line (P> 0.1) The level is higher than A549. CONCLUSION: PAα has high invasiveness to A549. Its mechanism may be related to the high expression and phosphorylation of EGFR, the high expression of tTG and MMP-2. TTG can regulate the level of MMP-2 but not itself by the EGFR signaling pathway Regulation.