Methanolic Extracts of Bitter Melon Inhibit Colon Cancer Stem Cells by Affecting Energy Homeostasis and Autophagy

Methanolic Extracts of Bitter Melon Inhibit Colon Cancer Stem Cells by Affecting Energy Homeostasis and Autophagy Deep Kwatra Department of Molecular and Integrative Physiology, University of Kansas Medical Center, 3901 Rainbow Boulevard MS 3040, Kansas City, KS 66160, USA Dharmalingam Subramaniam Department of Molecular and Integrative Physiology, University of Kansas Medical Center, 3901 Rainbow Boulevard MS 3040, Kansas City, KS 66160, USA University of Kansas Cancer Center, University of Kansas Medical Center, 3901 Rainbow Boulevard MS 3040, Kansas City, KS 66160, USA Prabhu Ramamoorthy Department of Molecular and Integrative Physiology, University of Kansas Medical Center, 3901 Rainbow Boulevard MS 3040, Kansas City, KS 66160, USA University of Kansas Cancer Center, University of Kansas Medical Center, 3901 Rainbow Boulevard MS 3040, Kansas City, KS 66160, USA David Standing Department of Molecular and Integrative Physiology, University of Kansas Medical Center, 3901 Rainbow Boulevard MS 3040, Kansas City, KS 66160, USA University of Kansas Cancer Center, University of Kansas Medical Center, 3901 Rainbow Boulevard MS 3040, Kansas City, KS 66160, USA Elizabeth Moran Department of Molecular and Integrative Physiology, University of Kansas Medical Center, 3901 Rainbow Boulevard MS 3040, Kansas City, KS 66160, USA Ravichandiran Velayutham Department of Pharmacognosy, Vels University, Pallavaram, Chennai 600117, India Ashim Mitra Department of Pharmaceutical Sciences, University of Missouri at Kansas City, Kansas City, MO 64108, USA Shahid Umar Department of Molecular and Integrative Physiology, University of Kansas Medical Center, 3901 Rainbow Boulevard MS 3040, Kansas City, KS 66160, USA University of Kansas Cancer Center, University of Kansas Medical Center, 3901 Rainbow Boulevard MS 3040, Kansas City, KS 66160, USA Shrikant Anant Department of Molecular and Integrative Physiology, University of Kansas Medical Center, 3901 Rainbow Boulevard MS 3040, Kansas City, KS 66160, USA University of Kansas Cancer Center, University of Kansas Medical Center, 3901 Rainbow Boulevard MS 3040, Kansas City, KS 66160, USA

Bitter melon fruit is recommended in ancient Indian and Chinese medicine for prevention/treatment of diabetes. However its effects on cancer progression are not well understood. Here, we have determined the efficacy of methanolic extracts of bitter melon on colon cancer stem and progenitor cells. Both, whole fruit (BMW) and skin (BMSk) extracts showed significant inhibition of cell proliferation and colony formation, with BMW showing greater efficacy. In addition, the cells were arrested at the S phase of cell cycle. Moreover, BMW induced the cleavage of LC3B but not caspase 3/7, suggesting that the cells were undergoing autophagy and not apoptosis. Further confirmation of autophagy was obtained when western blots showed reduced Bcl-2 and increased Beclin-1, Atg 7 and 12 upon BMW treatment. BMW reduced cellular ATP levels coupled with activation of AMP activated protein kinase; on the other hand, exogenous additions of ATP lead to revival of cell proliferation. Finally, BMW treatment results in a dose-dependent reduction in the number and size of colonospheres. The extracts also decreased the expression of DCLK1 and Lgr5, markers of quiescent, and activated stem cells. Taken together, these results suggest that the extracts of bitter melon can be an effective preventive/therapeutic agent for colon cancer.
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