Ju-Hee Kang1, Jeong-Eun Jang2, Siddhartha Kumar Mishra3, Hee-Ju Lee4, Chu Won Nho4, Dongyun Shin2, Mirim Jin5, Mi Kyung Kim6, Changsun Choi7, Seung Hyun Oh8. 1. Gachon Institute of Pharmaceutical Sciences, Gachon University, Incheon 406-840 Republic of Korea; National Cancer Center, Goyang-si, Gyeonggi-do 410-769, Republic of Korea; Department of Food and Nutrition, School of Food Science and Technology, Chung-Ang University, Anseong-si, Gyeonggi-do 456-756, Republic of Korea. 2. Gachon Institute of Pharmaceutical Sciences, Gachon University, Incheon 406-840 Republic of Korea. 3. Department of Zoology, School of Biological Sciences, Dr. Harisingh Gour Central University, Sagar 470003, India. 4. Korea Institute of Science and Technology (KIST), Gangneung Institute, Gangneung-si, Gangwon-do 210-340, Republic of Korea. 5. College of Korean Medicine, Daejeon University, Daejeon 300-716, Republic of Korea. 6. National Cancer Center, Goyang-si, Gyeonggi-do 410-769, Republic of Korea. 7. Department of Food and Nutrition, School of Food Science and Technology, Chung-Ang University, Anseong-si, Gyeonggi-do 456-756, Republic of Korea. 8. Gachon Institute of Pharmaceutical Sciences, Gachon University, Incheon 406-840 Republic of Korea. Electronic address: eyeball@hanmail.net.
Abstract
AIM OF THE STUDY: In this study, we examined the effect of different fractions and components of Chaga mushroom (Inonotus Obliquus) on viability and apoptosis of colon cancer cells. Among them, one component showed the most effective growth inhibition and was identified as ergosterol peroxide by NMR analysis. We investigated the anti-proliferative and apoptosis mechanisms of ergosterol peroxide associated with its anti-cancer activities in human colorectal cancer (CRC) cell lines and tested its anti-tumor effect on colitis-induced CRC developed by Azoxymethane (AOM)/Dextran sulfate sodium (DSS) in a mouse model. MATERIALS AND METHODS: We used MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays, flow cytometry assays, Western blot analysis, colony formation assays, reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemistry (IHC), and AOM/DSS mouse models to study the molecular mechanism of metastatic activities in CRC cells. RESULTS: Ergosterol peroxide inhibited cell proliferation and also suppressed clonogenic colony formation in HCT116, HT-29, SW620 and DLD-1 CRC cell lines. The growth inhibition observed in these CRC cell lines was the result of apoptosis, which was confirmed by FACS analysis and Western blotting. Ergosterol peroxide inhibited the nuclear levels of β-catenin, which ultimately resulted in reduced transcription of c-Myc, cyclin D1, and CDK-8. Ergosterol peroxide administration showed a tendency to suppress tumor growth in the colon of AOM/DSS-treated mice, and quantification of the IHC staining showed a dramatic decrease in the Ki67-positive staining and an increase in the TUNEL staining of colonic epithelial cells in AOM/DSS-treated mice by ergosterol peroxide for both prevention and therapy. CONCLUSION: Our data suggest that ergosterol peroxide suppresses the proliferation of CRC cell lines and effectively inhibits colitis-associated colon cancer in AOM/DSS-treated mice. Ergosterol peroxide down-regulated β-catenin signaling, which exerted anti-proliferative and pro-apoptotic activities in CRC cells. These properties of ergosterol peroxide advocate its use as a supplement in colon cancer chemoprevention.
AIM OF THE STUDY: In this study, we examined the effect of different fractions and components of Chaga mushroom (Inonotus Obliquus) on viability and apoptosis of colon cancer cells. Among them, one component showed the most effective growth inhibition and was identified as ergosterol peroxide by NMR analysis. We investigated the anti-proliferative and apoptosis mechanisms of ergosterol peroxide associated with its anti-cancer activities in humancolorectal cancer (CRC) cell lines and tested its anti-tumor effect on colitis-induced CRC developed by Azoxymethane (AOM)/Dextran sulfate sodium (DSS) in a mouse model. MATERIALS AND METHODS: We used MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays, flow cytometry assays, Western blot analysis, colony formation assays, reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemistry (IHC), and AOM/DSSmouse models to study the molecular mechanism of metastatic activities in CRC cells. RESULTS:Ergosterol peroxide inhibited cell proliferation and also suppressed clonogenic colony formation in HCT116, HT-29, SW620 and DLD-1 CRC cell lines. The growth inhibition observed in these CRC cell lines was the result of apoptosis, which was confirmed by FACS analysis and Western blotting. Ergosterol peroxide inhibited the nuclear levels of β-catenin, which ultimately resulted in reduced transcription of c-Myc, cyclin D1, and CDK-8. Ergosterol peroxide administration showed a tendency to suppress tumor growth in the colon of AOM/DSS-treated mice, and quantification of the IHC staining showed a dramatic decrease in the Ki67-positive staining and an increase in the TUNEL staining of colonic epithelial cells in AOM/DSS-treated mice by ergosterol peroxide for both prevention and therapy. CONCLUSION: Our data suggest that ergosterol peroxide suppresses the proliferation of CRC cell lines and effectively inhibits colitis-associated colon cancer in AOM/DSS-treated mice. Ergosterol peroxide down-regulated β-catenin signaling, which exerted anti-proliferative and pro-apoptotic activities in CRC cells. These properties of ergosterol peroxide advocate its use as a supplement in colon cancer chemoprevention.
Authors: Yhiya M Amen; Qinchang Zhu; Hai-Bang Tran; Mohamed S Afifi; Ahmed F Halim; Ahmed Ashour; Amira Mira; Kuniyoshi Shimizu Journal: J Nat Med Date: 2016-02-22 Impact factor: 2.343
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