Alpha 9-Ncotinic Acetylcholine Receptors mediate angiogenesis in triple negative breast cancers Sonjid Ochirbat (1), Tzu-Hsuan Huang (2), Tzu-Chun Kan (2), Jungshan Chang (1,2,3*)
1) International Master/Ph.D Program in Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan
2) Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei, Taiwan
3) International Ph.D Program for Cell Therapy and Regeneration Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan
Abstract
Triple-negative breast cancer (TNBC) remains a significant clinical challenge because of its high vascularity and metastatic rates. Due to lack of surface targeted molecules, chemotherapy is the major strategy for treatments of TNBC patients, contributing to adversary side effects. Previous epidemiological cohort studies demonstrated that both active and passive smoking are associated with risk factors for developing breast cancer. Recently, several studies found that nicotinic acetylcholine receptors ɑ-9 (ɑ-9-nAChRs) is highly expressed on breast tumors, especially tumors derived from patients diagnosed at advanced stage. In vitro studies, it demonstrated that activation of ɑ-9-nAChRs promote carcinogenic behavior including prolonging cell survival, proliferation, increased angiogenesis and metastasis. Therefore, we hypothesized that anti-ɑ-9-nAChRs antibodies can be a putative therapeutic drug for TNBC treatment. In this study, we evaluated therapeutic effect of new bispecific antibody with two antigen-binding sites against ɑ-9-nAChR and methoxy-polyethylene glycol (mPEG), which provides the linkage to commercial chemotherapy drug, liposomal doxorubicin (Lipo-Dox). Our results showed that anti-α-9 and anti-mPEG (α--α-9-α-PEG) bispecific antibody decreased nicotine-induced angiogenic ability on both HUVECs and cancer cells. To characterize molecular mechanism in cancer cells, α--α-9-α-PEG antibody inhibits nicotine induced angiogenesis through downregulating VEGF-A and its receptors (VEGFR2) expression. As well as, the impact of ɑ--ɑ-9-ɑ-PEG bispecific antibody was assessed on cancer cell motility. The nicotine-induced MDA-MB-231 cell migration and invasion were significantly decreased in a dose dependent manner by both transwell chamber assay and wound healing assay. Taken together, results showed that ɑ--ɑ-9-ɑ-PEG bispecific antibody inhibits TNBC progression through suppressing the cancer cell migration, invasion and angiogenesis. We suggest that ɑ--ɑ-9-ɑ-PEG bispecific antibody as
