Activation of HERV-K Env protein is essential for tumorigenesis and metastasis of breast cancer cells

// Fuling Zhou 1, 2, * , Ming Li 1, * , Yongchang Wei 1, 3, * , Kevin Lin 4 , Yue Lu 4 , Jianjun Shen 4 , Gary L. Johanning 1 , Feng Wang-Johanning 1 1 Viral Oncology Program, Center for Cancer and Metabolism, SRI International, Menlo Park, California, USA 2 Department of Clinical Hematology, Zhongnan Hospital, Wuhan University, Wuhan, Hubei, China 3 Department of Radiation and Medical Oncology, Zhongnan Hospital, Wuhan University, Wuhan, Hubei, China 4 Department of Epigenetics and Molecular Carcinogenesis, Science Park, The University of Texas MD Anderson Cancer Center, Smithville, Texas, USA * Co-first authors Correspondence to: Feng Wang-Johanning, email: feng.wang-johanning@SRI.com Keywords: HERV-K, breast cancer, shRNA, Ras signaling, tumorigenesis and metastasis Received: April 22, 2016 Accepted: August 11, 2016 Published: August 20, 2016 ABSTRACT Human endogenous retrovirus type K (HERV-K) Env protein was previously demonstrated to be overexpressed in human breast cancer (BC) cells and tissues. However, the molecular pathways driving the specific alterations are unknown. We now show that knockdown of its expression with an shRNA (shRNAenv) blocked BC cell proliferation, migration, and invasion. shRNAenv transduction also attenuated the ability of BC cells to form tumors, and notably prevented metastasis. Mechanistically, downregulation of HERV-K blocked expression of tumor-associated genes that included Ras, p-RSK, and p-ERK. The major upstream regulators influenced by HERV-K knockdown were p53, TGF- β1, and MYC. Of interest, when the HERV-K env gene was overexpressed in shRNAenv-transduced BC cells using an HERV-K env expression vector, Ras/Raf/MEK/ERK pathway signaling was restored. CDK5, which alters p53 phosphorylation in some cancers, was upregulated and p53 was downregulated when HERV-K was overexpressed. CDK5 is also a mediator of TGF-β1-induced epithelial-mesenchymal transition and migration in cancer cells, and is involved in tumor formation. Importantly, reductions in migration, invasion, and transformation of BC cells stably transduced with shRNAenv was reversed after adding back a vector with a synonymous mutation of HERV-K env . Taken together, these results indicate that HERV-K Env protein plays an important role in tumorigenesis and metastasis of BC.