Specifically targeting ERK1 or ERK2 kills Melanoma cells

Journal of Translational Medicine

Brian J Nickoloff³  Hong Xin¹  Jianzhong Qin² 

[1] Department of Obstetrics and Gynecology, Northwestern University School of Medicine Feinberg School of Medicine, Chicago, IL 60611, USA;Department of Microbiology and Immunology, University of Illinois at Chicago Medical Center, Chicago, IL 60612, USA;Center for Cancer Genomics and Computational Biology, Laboratory of Cutaneous Oncology, Van Andel Research Institute, 333 Bostwick Ave NE, Room 5008, Grand Rapids, MI 49503-2518, USA

关键词: PLX4032;    BRAF;    Drug resistance;    Melanoma;    ERK;   

Others  :  1207780
DOI  :  10.1186/1479-5876-10-15

 received in 2012-01-09, accepted in 2012-01-25,  发布年份 2012

【 摘 要 】

Background

Overcoming the notorious apoptotic resistance of melanoma cells remains a therapeutic challenge given dismal survival of patients with metastatic melanoma. However, recent clinical trials using a BRAF inhibitor revealed encouraging results for patients with advanced BRAF mutant bearing melanoma, but drug resistance accompanied by recovery of phospho-ERK (pERK) activity present challenges for this approach. While ERK1 and ERK2 are similar in amino acid composition and are frequently not distinguished in clinical reports, the possibility they regulate distinct biological functions in melanoma is largely unexplored.

Methods

Rather than indirectly inhibiting pERK by targeting upstream kinases such as BRAF or MEK, we directly (and near completely) reduced ERK1 and ERK2 using short hairpin RNAs (shRNAs) to achieve sustained inhibition of pERK1 and/or pERK2.

Results and discussion

Using A375 melanoma cells containing activating BRAF^V600E mutation, silencing ERK1 or ERK2 revealed some differences in their biological roles, but also shared roles by reduced cell proliferation, colony formation in soft agar and induced apoptosis. By contrast, chemical mediated inhibition of mutant BRAF (PLX4032) or MEK (PD0325901) triggered less killing of melanoma cells, although they did inhibit proliferation. Death of melanoma cells by silencing ERK1 and/or ERK2 was caspase dependent and accompanied by increased levels of Bak, Bad and Bim, with reduction in p-Bad and detection of activated Bax levels and loss of mitochondrial membrane permeability. Rare treatment resistant clones accompanied silencing of either ERK1 and/or ERK2. Unexpectedly, directly targeting ERK levels also led to reduction in upstream levels of BRAF, CRAF and pMEK, thereby reinforcing the importance of silencing ERK as regards killing and bypassing drug resistance.

Conclusions

Selectively knocking down ERK1 and/or ERK2 killed A375 melanoma cells and also increased the ability of PLX4032 to kill A375 cells. Thus, a new therapeutic window is open for future clinical trials in which agents targeting ERK1 and ERK2 should be considered in patients with melanoma.

【 授权许可】

   
2012 Qin et al; BioMed Central Ltd.

【 图 表 】

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