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。
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 PAn 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 PAn was 165.3 times of A549 （.ells （P 〈 0.05）. The transcription levels of tTG and EGFR mRNA in PAot 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 PAn cells than in A549 （＇.ells. There was no significant difference in MMP-2 mRNA transcriplion level between A549 and PAa cells （P 〉 0. 1 ）, but the expression level of MMP-2 protein was higher in PAot ceils than in A549 cells. Conclusions PAot 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.
出处
《实用医学杂志》
CAS
北大核心
2013年第21期3472-3474,共3页
The Journal of Practical Medicine
基金
广州市重点学科项目(编号:B127007)
广州市科技计划项目(编号:2012J4100009)
