Role of PAX8 in the regulation of MET and RON receptor tyrosine kinases in non-small cell lung cancer

BMC Cancer

Ravi Salgia²  Everett E Vokes³  Jay Sharma⁵  Sherven Sharma⁴  Aliya N Husain¹  Maria Tretiakova¹  Immanuel Dhanasingh³  Essam El-Hashani³  Rajani Kanteti³ 

[1]Department of Pathology, University of Chicago Medical Center, Chicago, IL, USA

[2]University of Chicago, Department of Medicine, 5841 S. Maryland Avenue, Chicago, IL 60637, USA

[3]Department of Hematology/Oncology, University of Chicago Medical Center, Chicago, IL, USA

[4]Department of Medicine, University of California, Los Angeles, CA, USA

[5]Department of stem cell Research, Cellprogen Inc, San Pedro, CA, USA

关键词: NSCLC;    RON;    MET;    PAX8;   

Others  :  858971
DOI  :  10.1186/1471-2407-14-185

 received in 2013-08-01, accepted in 2014-02-28,  发布年份 2014

【 摘 要 】

Background

Non-small cell lung cancers (NSCLC) are highly heterogeneous at the molecular level and comprise 75% of all lung tumors. We have previously shown that the receptor tyrosine kinase (RTK) MET frequently suffers gain-of-function mutations that significantly promote lung tumorigenesis. Subsequent studies from our lab also revealed that PAX5 transcription factor is preferentially expressed in small cell lung cancer (SCLC) and promotes MET transcription. PAX8, however, is also expressed in NSCLC cell lines. We therefore investigated the role of PAX8 in NSCLC.

Methods

Using IHC analysis, PAX8 protein expression was determined in archival NSCLC tumor tissues (n = 254). In order to study the effects of PAX8 knockdown on NSCLC cellular functions such as apoptosis and motility, siRNA against PAX8 was used. Confocal fluorescence microscopy was used to monitor the localization of MET, RON and PAX8. The combinatorial effect of PAX8 knockdown and MET inhibition using SU11274 was investigated in NSCLC cell viability assay.

Results

Relative levels of PAX8 protein were elevated (≥ + 2 on a scale of 0–3) in adenocarcinoma (58/94), large cell carcinoma (50/85), squamous cell carcinoma (28/47), and metastatic NSCLC (17/28; lymph node). Utilizing early progenitors isolated from NSCLC cell lines and fresh tumor tissues, we observed robust overexpression of PAX8, MET, and RON. PAX8 knockdown A549 cells revealed abrogated PAX8 expression with a concomitant loss in MET and the related RON kinase expression. A dramatic colocalization between the active form of MET (also RON) and PAX8 upon challenging A549 cells with HGF was visualized. A similar colocalization of MET and EGL5 (PAX8 ortholog) proteins was found in embryos of C. elegans. Most importantly, knockdown of PAX8 in A549 cells resulted in enhanced apoptosis (~6 fold) and decreased cell motility (~45%), thereby making PAX8 a potential therapeutic target. However, the combinatorial approach of PAX8 knockdown and treatment with MET inhibitor, SU11274, had marginal additive effect on loss of NSCLC cell viability.

Conclusion

PAX8 provides signals for growth and motility of NSCLC cells and is necessary for MET and RON expression. Further investigations are necessary to investigate the therapeutic potential of PA8 in NSCLC.

【 授权许可】

   
2014 Kanteti et al.; licensee BioMed Central Ltd.

【 图 表 】

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