Alternative Therapeutic Approach to Urothelial Cell Carcinoma with Medicinal Mushroom Extracts

Amul Bhalodi¹, Mark Ferretti¹, Matthew Chaimowitz¹, Muhammad Choudhury¹, John Phillips¹ and Sensuke Konno^1,

¹Department of Urology, New York Medical College, Valhalla, New York, USA

Cite this paper:
Amul Bhalodi, Mark Ferretti, Matthew Chaimowitz, Muhammad Choudhury, John Phillips and Sensuke Konno. Alternative Therapeutic Approach to Urothelial Cell Carcinoma with Medicinal Mushroom Extracts. Journal of Cancer Research and Treatment. 2016; 4(5):73-79. doi: 10.12691/jcrt-4-5-1

Abstract

Severe side effects from chemotherapy or immunotherapy often limit their clinical utility in urothelial cell carcinoma of the bladder. To explore alternative therapies, we were particularly interested in one of medicinal mushrooms called Phellinus linteus (PL) with antitumor/anticancer activity. We investigated possible anticancer effects of three distinct extracts or fractions of PL using an urothelial cell carcinoma (UCC) model in vitro. Those fractions included PL-I (crude extract), PL-II (water-extracted), and PL-III (ethanol-extracted), which were tested for their antiproliferative effects on UCC cells. To explore the anticancer mechanism(s) of these fractions, cell cycle analysis and analyses of epigenetic parameters, caspases-3 and -9, and apoptotic regulators, were also performed. Both PL-I and -II induced a maximum growth reduction of ~60% at 700 µg/ml while PL-III led to a ~90% growth reduction at 150 µg/ml in 72 h. Cell cycle analysis indicated that cells treated with 500 µg/ml (IC₅₀) of PL-I and -II or 60 µg/ml (IC₅₀) of PL-III underwent a G₁ cell cycle arrest, accompanied by the up-regulation of p21, a cell cycle-dependent kinase inhibitor. Additionally, histone deacetylase (HDAC) activity was significantly (>60%) lost, while both histones H3 and H4 were highly acetylated, indicating alterations in the chromatin structure. Moreover, both caspases-3 and -9 in cells treated with each IC₅₀ of three PL-fractions were all significantly (p<0.03) activated, implying induction of apoptosis. The present study shows that all three PL-fractions, PL-I, -II, and -III, have anticancer effects on UCC cells, although PL-III appears to be the most potent. Such an anticancer mechanism is attributed to a p21-mediated cell cycle arrest with epigenetic modifications involving HDAC inactivation and hyperacetylation of H3 and H4, ultimately leading to apoptotic cell death. Therefore, PL-fractions may have clinical implications in a safer and improved therapeutic modality for urothelial cell carcinoma.

Keywords:
mushrooms Phellinus linteus cell cycle histones urothelial cell carcinoma

This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

References:

[1]	Ferlay J, Soerjomataram I, Dikshit R, et al. Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer 2015; 136(5): E359-86.
[2]	Siegel R, Ma J, Zou Z, Jemal A. Cancer statistics 2014. CA Cancer J Clin 2014; 64(1): 9-29.
[3]	Kaufman DS, Shipley WU, Feldman AS. Bladder cancer. Lancet 2009; 374(9685): 239-49.
[4]	Grossman HB, Natale RB, Tangen CM, et al. Neoadjuvant chemotherapy plus cystectomy compared with cystectomy alone for locally advanced bladder cancer. N Engl J Med 2003; 349(9): 859-66.
[5]	Zhu T, Kim SH, Chen CY. A medicinal mushroom: Phellinus linteus. Curr Med Chem 2008; 15(13): 1330-5.
[6]	Sliva D. Medicinal mushroom Phellinus linteus as an alternative cancer therapy. Exp Ther Med 2010; 1(3): 407-11.
[7]	Kim GY, Oh YH, Park YM. Acidic polysaccharide isolated from Phellinus linteus induces nitric oxide-mediated tumoricidal activity of macrophages through protein tyrosine kinase and protein kinase C. Biochem Biophys Res Commun 2003; 309(2): 399-407.
[8]	Lee YS, Kim YH, Shin EK, et al. Anti-angiogenic activity of methanol extract of Phellinus linteus and its fractions. J Ethnopharmacol 2010; 131(1): 56-62.
[9]	Lee JW, Baek SJ, Bae WC, Park JM, Kim YS. Antitumor and antioxidant activities of the extracts from fruiting body of Phellinus linteus. Mycobiology 2006; 34(4): 230-5.
[10]	Zhu T, Guo J, Collins L, et al. Phellinus linteus activates different pathways to induce apoptosis in prostate cancer cells. Br J Cancer 2007; 96(4): 583-90.
[11]	Tsuji T, Du W, Nishioka T, Chen L, Yamamoto D, Chen CY. Phellinus linteus extract sensitizes advanced prostate cancer cells to apoptosis in athymic nude mice. PLoS One 2010; 5(3): e9885.
[12]	Guo J, Zhu T, Collins L, Xiao ZX, Kim SH, Chen CY. Modulation of lung cancer growth arrest and apoptosis by Phellinus linteus. Mol Carcinog 2007; 46(2): 144-54.
[13]	Yang BK, Hwang SL, Yun IJ, et al. Antitumor effects and immunomodulating activities of Phellinus linteus extract in a CT-26 cell-injected colon cancer mouse model. Mycobiology 2009; 37(2): 128-32.
[14]	Sliva D, Jedinak A, Kawasaki J, Harvey K, Sliva V. Phellinus linteus suppresses growth, angiogenesis and invasive behaviour of breast cancer cells through the inhibition of AKT signaling. Br J Cancer 2008; 98(8): 1348-56.
[15]	Chen YS, Lee SM, Lin CC, Liu CY. Hispolon decreases melanin production and induces apoptosis in melanoma cells through the downregulation of tyrosinase and microphthalmia-associated transcription factor (MITF) expressions and the activation of caspase-3, -8 and -9. Int J Mol Sci 2014; 15(1): 1201-15.
[16]	Chen YC, Chang HY, Deng JS, et al. Hispolon from Phellinus linteus induces G0/G1 cell cycle arrest and apoptosis in NB4 human leukemia cells. Am J Chin Med 2013; 41(6): 1439-57.
[17]	Tang Y, Simoneau AR, Xie J, Shahandeh B, Zi X. Effects of the kava chalcone flavokawain a differ in bladder cancer cells with wild-type versus mutant p53. Cancer Prev Res (Phila) 2008; 1(6): 439-51.
[18]	Sherr CJ. The Pezcoller lecture: cancer cell cycle revisited. Cancer Res 2000; 60(14): 3689-95.
[19]	Sherr CJ, Roberts JM. CDK inhibitors: positive and negative regulators of G1-phase progression. Genes Dev 1999; 13(12): 1501-12.
[20]	Richon VM, Sandhoff TW, Rifkind RA, Marks PA. Histone deacetylase inhibitor selectively induces p21WAF1 expression and gene-associated histone acetylation. Proc Natl Acad Sci USA 2000; 97(18): 10014-9.
[21]	Villar-Garea A, Esteller M. Histone deacetylase inhibitors: understanding a new wave of anticancer agents. Int J Cancer 2004; 112(2): 171-8.
[22]	de Ruijter AJ, van Gennip AH, Caron HN, Kemp S, van Kuilenburg AB. Histone deacetylases (HDACs): characterization of the classical HDAC family. Biochem J 2003; 370(Pt 3): 737-49.
[23]	Pop C, Salvesen GS. Human caspases: activation, specificity, and regulation. J Biol Chem 2009; 284(33): 21777-81.
[24]	Fiandalo MV, Kyprianou N: Caspase control: protagonists of cancer cell apoptosis. Exp Oncol 2012; 34(3): 165-75.
[25]	Li YG, Ji DF, Zhong S et al: Polysaccharide from Phellinus linteus induces S-phase arrest in HepG2 cells by decreasing calreticulin expression and activating the p27kip1-cyclin A/D1/E-CDK2 pathway. J Ethnopharmacol 2013; 150(1): 187-95.
[26]	Zhong S, Ji DF, Li YG, Lin TB, Lv ZQ, Chen HP. Activation of p27kip1-cyclin D1/E-CDK2 pathway by polysaccharide from Phellinus linteus leads to S-phase arrest in HT-29 cells. Chem Biol Interact 2013; 206(2): 222-9.
[27]	Harb-de la Rosa A, Acker M, Kumar RA, Manoharan M. Epigenetics application in the diagnosis and treatment of bladder cancer. Can J Urol 2015; 22(5): 7947-51.
[28]	Park HJ, Choi SY, Hong SM, Hwang SG, Park DK. The ethyl acetate extract of Phellinus linteus grown on germinated brown rice induces G0/G1 cell cycle arrest and apoptosis in human colon carcinoma HT-29 cells. Phytother Res 2010; 24(7): 1019-26.
[29]	Lim JH, Lee YM, Park SR, Kim DH, Lim BO. Anticancer activity of hispidin via reactive oxygen species-mediated apoptosis in colon cancer cells. Anticancer Res 2014; 34(8): 4087-93.
[30]	Li YG, Ji DF, Zhong S, Zhu JX, Chen S, Hu GY. Anti-tumor effects of proteoglycan from Phellinus linteus by immunomodulating and inhibiting Reg IV/EGFR/Akt signaling pathway in colorectal carcinoma. Int J Biol Macromol 2011; 48(3): 511-7.