Monosaccharide digitoxin derivative sensitize human non-small cell lung cancer cells to anoikis through Mcl-1 proteasomal degradation.
Advanced stage cancers acquire anoikis resistance which provides metastatic potential to invade and form tumors at distant sites. Suppression of anoikis resistance by novel molecular therapies would greatly benefit treatment strategies for metastatic cancers. Recently, digitoxin and several of its novel synthetic derivatives, such as α-l-rhamnose monosaccharide derivative (D6-MA), have been synthesized and studied for their profound anticancer activity in various cancer cell lines. In this study, we investigated the anoikis sensitizing effect of D6-MA compared with digitoxin to identify their anti-metastatic mechanism of action. D6-MA sensitized NSCLC H460 cells to detachment-induced apoptosis with significantly greater cytotoxicity (IC50=11.9 nM) than digitoxin (IC50=90.7 nM) by activating caspase-9. Screening of the Bcl-2 protein family revealed that degradation of anti-apoptotic Mcl-1 protein is a favorable target. Mcl-1 over-expression and knockdown studies in D6-MA and digitoxin exposed cells resulted in rescue and enhancement, respectively, indicating a facilitative role for decreased Mcl-1 expression in NSCLC anoikis. Transfection with mutant Mcl-1S159 attenuated detachment-induced cell death and correlated with a remaining of Mcl-1 level. Furthermore, D6-MA suppressed Mcl-1 expression via ubiquitin proteasomal degradation that is dependent on activation of glycogen synthase kinase (GSK)-3β signaling. In addition, D6-MA also targeted Mcl-1 degradation causing an increased anoikis in A549 lung cancer cells. Anoikis sensitizing effect on normal small airway epithelial cells was not observed indicating the specificity of D6-MA and digitoxin for NSCLC. These results identify a novel cardiac glycoside (CG) sensitizing anoikis mechanism and provide a promising anti-metastatic target for lung cancer therapy.
Cell-Based Drug and Health Product Development Research Unit, Department of Pharmacology and Physiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand.|Department of Basic Pharmaceutical Sciences, West Virginia University, Morgantown, WV 26506, United States.
Varisa
V
Pongrakhananon
Department of Basic Pharmaceutical Sciences, West Virginia University, Morgantown, WV 26506, United States.|Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Morgantown, WV 26505, United States.
Todd A
TA
Stueckle
Department of Chemistry and Chemical Biology, Northeastern University, Boston, MA 02115, United States.
Hua-Yu Leo
HL
Wang
Department of Chemistry and Chemical Biology, Northeastern University, Boston, MA 02115, United States.
George A
GA
O'Doherty
Department of Chemical Engineering, West Virginia University, Morgantown, WV 26506, United States.
Cerasela Zoica
CZ
Dinu
Cell-Based Drug and Health Product Development Research Unit, Department of Pharmacology and Physiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand.
Pithi
P
Chanvorachote
Department of Basic Pharmaceutical Sciences, West Virginia University, Morgantown, WV 26506, United States.|Mary Babb Randolph Cancer Center, West Virginia University, Morgantown, WV 26506, United States.
Yon
Y
Rojanasakul
D023102:Anoikis / Q000187:drug effects*; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D002289:Carcinoma, Non-Small-Cell Lung / Q000378:metabolism / Q000473:pathology; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D004074:Digitoxin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D008175:Lung Neoplasms / Q000378:metabolism / Q000473:pathology; D064549:Myeloid Cell Leukemia Sequence 1 Protein / Q000037:antagonists & inhibitors* / Q000096:biosynthesis
D000970:Antineoplastic Agents; C587796:D6-MA compound; C579095:MCL1 protein, human; D064549:Myeloid Cell Leukemia Sequence 1 Protein; D004074:Digitoxin
Anoikis; Glycogen synthase kinase-3β; Mcl-1; Metastasis; Monosaccharide digitoxin derivative; NSCLC