CM  Vol.4 No.2 , June 2013
The Effects of Ganoderma lucidum on Initial Events Related to the Bacillus Calmette-Guérin Efficacy and Toxicity on High-Risk Uroepithelial Cells: An in Vitro Preliminary Study
Abstract: A novel prophylactic regimen is demanded for preventing bladder cancer recurrence, because of the high side-effect tolls of conventional adjuvant Bacillus Calmette-Guérin (BCG) immunotherapy, in addition to its only moderate efficacy. In vitro and animal studies have demonstrated the anti-cancer properties of a medicinal mushroom called Ganoderma lucidum (GL). In this study, a pre-malignant human uroepithelial cells (HUC-PC) model was utilized to compare the effectiveness between ethanol extract of GL (GLe) and BCG on interleukin-6 (IL-6) secretion and lactate dehydrogenase (LDH) cytotoxicity. Additionally, parameters relevant to the BCG efficacy and safety, including free soluble fibronectin (FN) and cell-surface glycosaminoglycans (GAGs) levels were tested, following the exposure of GLe to the cells. GLe at 100 μg/ml and BCG at 4.8 × 107 CFU were shown to induce equivalent levels of IL-6, suggesting the potential synergism, while the tested concentrations of GLe were non-cytotoxic. During the initial four hours of GLe exposure, the free FN concentrations in harvested media were significantly reduced that might facilitate the binding of BCG for uroepithelial internalization to enhance BCG efficacy. Furthermore, the cell membrane-bound GAGs levels of HUC-PC cells were significant increased in response to GLe to suggest cellular protection from BCG infection. In summary, current findings suggest the potential additive synergism of GLe with the BCG efficacy, as well as its protective effects, and thus reducing the BCG toxicity.
Cite this paper: J. Yuen, M. Gohel and C. Ng, "The Effects of Ganoderma lucidum on Initial Events Related to the Bacillus Calmette-Guérin Efficacy and Toxicity on High-Risk Uroepithelial Cells: An in Vitro Preliminary Study," Chinese Medicine, Vol. 4 No. 2, 2013, pp. 52-61. doi: 10.4236/cm.2013.42009.
References

[1]   Q. Y. Lu, Y. S. Jin, Q. Zhang, Z. Zhang, D. Heber, V. L. Go, et al., “Ganoderma lucidum Extracts Inhibit Growth and Induce Actin Polymerization in Bladder Cancer Cells in Vitro,” Cancer Letters, Vol. 216, No. 1, 2004, pp. 9-20. doi:10.1016/j.canlet.2004.06.022

[2]   J. W. Yuen, M. D. Gohel and D. W. Au, “Telomerase-Associated Apoptotic Events by Mushroom Ganoderma Lucidum on Premalignant Human Urothelial Cells,” Nutrition and Cancer, Vol. 60, No. 1, 2008, pp. 109-119. doi:10.1080/01635580701525869

[3]   J. W. Yuen and M. D. Gohel, “The Dual Roles of Ganoderma Antioxidants on Urothelial Cell DNA under Carcinogenic Attack,” Journal of Ethnopharmacology, Vol. 118, No. 2, 2008, pp. 324-330. doi:10.1016/j.jep.2008.05.003

[4]   J. W. Yuen, M. D. Gohel and C. F. Ng, “The Differential Immunological Activities of Ganoderma lucidum on Human Pre-Cancerous Uroepithelial Cells,” Journal of Ethnopharmacology, Vol. 135, No. 1, 2011, pp. 711-718. doi:10.1016/j.jep.2011.04.005

[5]   W. Oosterlinck, “The Management of Superficial Bladder Cancer,” BJU International, Vol. 87, No. 2, 2001, pp. 135-140. doi:10.1046/j.1464-410x.2001.00948.x

[6]   J. T. Leppert, O. Shvarts, K. Kawaoka, R. Lieberman, A. S. Belldegrun and A. J. Pantuck, “Prevention of Bladder Cancer: A Review,” European Urology, Vol. 49, No. 2, 2006, pp. 226-234. doi:10.1016/j.eururo.2005.12.011

[7]   P. Tyagi, P. C. Wu, M. Chancellor, N. Yoshimura and L. Huang, “Recent Advances in Intravesical Drug/Gene Delivery,” Molecular Pharmaceutics, Vol. 3, No. 4, 2006, pp. 369-379. doi:10.1021/mp060001j

[8]   D. L. Lamm, W. R. McGee and K. Hale, “Bladder Cancer: Current Optimal Intravesical Treatment,” Urologic Nursing, Vol. 25, No. 5, 2005, pp. 323-326, 331-332.

[9]   K. Taniguchi, S. Koga, M. Nishikido, S. Yamashita, T. Sakuragi, H. Kanetake, et al., “Systemic Immune Response after Intravesical Instillation of Bacille Calmette-Guerin (BCG) for Superficial Bladder Cancer,” Clinical & Experimental Immunology, Vol. 115, No. 1, 1999, pp. 131135. doi:10.1046/j.1365-2249.1999.00756.x

[10]   M. Sanchez-Carbayo, M. Urrutia, R. Romani, M. Herrero, J. M. Gonzalez de Buitrago and J. A. Navajo, “Serial Urinary IL-2, IL-6, IL-8, TNF Alpha, UBC, CYFRA 21-1 and NMP22 during Follow-Up of Patients with Bladder Cancer Receiving Intravesical BCG,” Anticancer Research, Vol. 21, No. 4B, 2001, pp. 3041-3047.

[11]   J. J. Patard, F. Saint, F. Velotti, C. C. Abbou and D. K. Chopin, “Immune Response Following Intravesical Bacillus Calmette-Guerin Instillations in Superficial Bladder Cancer: A Review,” Urological Research, Vol. 26, No. 3, 1998, pp. 155-159. doi:10.1007/s002400050039

[12]   T. M. de Reijke, E. C. de Boer, K. H. Kurth and D. H. Schamhart, “Urinary Cytokines during Intravesical Bacillus Calmette-Guerin Therapy for Superficial Bladder Cancer: Processing, Stability and Prognostic Value,” Journal of Urology, Vol. 155, No. 2, 1996, pp. 477-482. doi:10.1016/S0022-5347(01)66424-3

[13]   T. M. de Reijke, P. C. Vos, E. C. de Boer, R. F. Bevers, W. H. de Muinck Keizer, K. H. Kurth, et al., “Cytokine Production by the Human Bladder Carcinoma Cell Line T24 in the Presence of Bacillus Calmette-Guerin (BCG),” Urological Research, Vol. 21, No. 5, 1993, pp. 349-352. doi:10.1007/BF00296835

[14]   K. Esuvaranathan, A. B. Alexandroff, M. McIntyre, A. M. Jackson, S. Prescott, G. D. Chisholm, et al., “Interleukin6 Production by Bladder Tumors Is Upregulated by BCG Immunotherapy,” Journal of Urology, Vol. 154, No. 2, 1995, pp. 572-575. doi:10.1016/S0022-5347(01)67113-1

[15]   Y. Zhang, R. Mahendran, L. L. Yap, K. Esuvaranathan and H. E. Khoo, “The Signalling Pathway for BCG-Induced Interleukin-6 Production in Human Bladder Cancer Cells,” Biochemical Pharmacology, Vol. 63, No. 2, 2002, pp. 273-282. doi:10.1016/S0006-2952(01)00831-0

[16]   R. F. Bevers, E. C. de Boer, K. H. Kurth and D. H. Schamhart, “BCG-Induced Interleukin-6 Upregulation and BCG Internalization in Well and Poorly Differentiated Human Bladder Cancer Cell Lines,” European Cytokine Network, Vol. 9, No. 2, 1998, pp. 181-186.

[17]   R. F. Bevers, K. H. Kurth and D. H. Schamhart, “Role of Urothelial Cells in BCG Immunotherapy for Superficial Bladder Cancer,” British Journal of Cancer, Vol. 91, No. 4, 2004, pp. 607-612.

[18]   L. R. Kavoussi, E. J. Brown, J. K. Ritchey and T. L. Ratliff, “Fibronectin-Mediated Calmette-Guerin Bacillus Attachment to Murine Bladder Mucosa. Requirement for the Expression of an Antitumor Response,” The Journal of Clinical Investigation, Vol. 85, No. 1, 1990, pp. 62-67. doi:10.1172/JCI114434

[19]   T. L. Ratliff, J. O. Palmer, J. A. McGarr and E. J. Brown, “Intravesical Bacillus Calmette-Guerin Therapy for Murine Bladder Tumors: Initiation of the Response by Fibronectin-Mediated Attachment of Bacillus Calmette-Guerin,” Cancer Research, Vol. 47, 1987, pp. 1762-1766.

[20]   J. Aslanzadeh, E. J. Brown, S. P. Quillin, J. K. Ritchey and T. L. Ratliff, “Characterization of Soluble Fibronectin Binding to Bacille Calmette-Guerin,” Journal of General Microbiology, Vol. 135, No. 10, 1989, pp. 2735-2741.

[21]   J. Miyazaki, K. Kawai, T. Oikawa, A. Johraku, K. Hattori, T. Shimazui, et al., “Uroepithelial Cells Can Directly Respond to Mycobacterium Bovis Bacillus Calmette-Guerin through Toll-Like Receptor Signaling,” BJU International, Vol. 97, No. 4, 2006, pp. 860-864. doi:10.1111/j.1464-410X.2006.06026.x

[22]   G. Zhang, F. Chen, Y. Xu, Y. Cao, S. Crist, A. McKerrow, et al., “Autocrine over Expression of Fibronectin by Human Transitional Carcinoma Cells Impairs Bacillus Calmette-Guerin Adherence and Signaling,” Journal of Urology, Vol. 172, No. 4, 2004, pp. 1496-1500. doi:10.1097/01.ju.0000140193.95528.91

[23]   Z. Akbulut, A. E. Canda, A. F. Atmaca, H. I. Cimen, C. Hasanoglu and M. D. Balbay, “BCG Sepsis Following Inadvertent Intravenous BCG Administration for the Treatment of Bladder Cancer Can Be Effectively Cured with Anti-Tuberculosis Medications,” The New Zealand Medical Journal, Vol. 123, No. 1325, 2010, pp. 72-77.

[24]   D. H. Schamhart, E. C. De Boer, R. Vleeming and K. H. Kurth, “Theoretical and Experimental Evidence on the Use of Glycosaminoglycans in BCG-Mediated Immunotherapy of Superficial Bladder Cancer,” Seminars in Thrombosis and Hemostasis, Vol. 20, No. 3, 1994, pp. 301-309. doi:10.1055/s-2007-1001917

[25]   D. H. Schamhart, E. C. de Boer and K. H. Kurth, “Interaction between Bacteria and the Lumenal Bladder Surface: Modulation by Pentosan Polysulfate, an Experimental and Theoretical Approach with Clinical Implication,” World Journal of Urology, Vol. 12, No. 1, 1994, pp. 27-37. doi:10.1007/BF00182048

[26]   F. Chen, G. Zhang, Y. Iwamoto and W. A. See, “BCG Directly Induces Cell Cycle Arrest in Human Transitional Carcinoma Cell Lines as a Consequence of Integrin Cross-Linking,” BMC Urology, Vol. 5, 2005, p. 8. doi:10.1186/1471-2490-5-8

[27]   Y. Zhang, H. E. Khoo and K. Esuvaranatha, “Effects of Bacillus Calmette-Guerin and Interferon-Alpha-2B on Human Bladder Cancer in Vitro,” International Journal of Cancer, Vol. 71, No. 5, 1997, pp. 851-857. doi:10.1002/(SICI)1097-0215(19970529)71:5<851::AID-IJC25>3.0.CO;2-9

[28]   R. W. Farndale, C. A. Sayers and A. J. Barrett, “A Direct Spectrophotometric Microassay for Sulfated Glycosaminoglycans in Cartilage Cultures,” Connective Tissue Research, Vol. 9, No. 4, 1982, pp. 247-248. doi:10.3109/03008208209160269

[29]   R. W. Farndale, D. J. Buttle and A. J. Barrett, “Improved Quantitation and Discrimination of Sulphated Glycosaminoglycans by Use of Dimethylmethylene Blue,” Biochimica et Biophysica Acta, Vol. 883, No. 2, 1986, pp. 173-177. doi:10.1016/0304-4165(86)90306-5

[30]   W. T. Chung, S. H. Lee, J. D. Kim, Y. S. Park, B. Hwang, S. Y. Lee, et al., “Effect of Mycelial Culture Broth of Ganoderma lucidum on the Growth Characteristics of Human Cell Lines,” Journal of Bioscience and Bioengineering, Vol. 92, No. 6, 2001, pp. 550-555.

[31]   H. S. Chen, Y. F. Tsai, S. Lin, C. C. Lin, K. H. Khoo, C. H. Lin, et al., “Studies on the Immuno-Modulating and Anti-Tumor Activities of Ganoderma lucidum (Reishi) Polysaccharides,” Bioorganic & Medicinal Chemistry, Vol. 12, No. 21, 2004, pp. 5595-5601.

[32]   F. Chen, P. Langenstroer, G. Zhang, Y. Iwamoto and W. A. See, “Androgen Dependent Regulation of Bacillus Calmette-Guerin Induced Interleukin-6 Expression in Human Transitional Carcinoma Cell Lines,” Journal of Urology, Vol. 170, No. 5, 2003, pp. 2009-2013.

[33]   F. Chen, G. Zhang, Y. Iwamoto and W. A. See, “Bacillus Calmette-Guerin Initiates Intracellular Signaling in a Transitional Carcinoma Cell Line by Cross-Linking Alpha 5 Beta 1 Integrin,” Journal of Urology, Vol. 170, No. 2, 2003, pp. 605-610.

[34]   P. Musiani, A. Modesti, M. Giovarelli, F. Cavallo, M. P. Colombo, P. L. Lollini, et al., “Cytokines, Tumour-Cell Death and Immunogenicity: A Question of Choice,” Immunology Today, Vol. 18, No. 1, 1997, pp. 32-36.

[35]   A. Sasaki, S. Kudoh, K. Mori, N. Takahashi and T. Suzuki, “Are BCG Effects against Urinary Bladder Carcinoma Cell Line T24 Correlated with Apoptosis in Vitro?” Urology International, Vol. 59, No. 3, 1997, pp. 142-148.

[36]   E. C. Borden, D. S. Groveman, T. Nasu, C. Reznikoff and G. T. Bryan, “Antiproliferative Activities of Interferons against Human Bladder Carcinoma Cell Lines in Vitro,” The Journal of Urology, Vol. 132, No. 4, 1984, pp. 800-803.

[37]   A. M. Jackson, A. B. Alexandroff, D. Fleming, S. Prescott, G. D. Chisholm and K. James, “Bacillus CalmetteGuerin (BCG) Organisms Directly Alter the Growth of Bladder Tumour Cells,” International Journal of Oncology, Vol. 5, No. 3, 1994, pp. 697-703.

[38]   International Agency for Research on Cancer, “Monographs on the Evaluation of Carcinogenic Risk of Chemicals,” World Health Organization, Geneva, 1972.

[39]   S. Y. Wang, M. L. Hsu, H. C. Hsu, C. H. Tzeng, S. S. Lee, M. S. Shiao, et al., “The Anti-Tumor Effect of Ganoderma Lucidum Is Mediated by Cytokines Released from Activated Macrophages and T Lymphocytes,” International Journal of Cancer, Vol. 70, No. 6, 1997, pp. 699-705. doi:10.1002/(SICI)1097-0215(19970317)70:6<699::AID-IJC12>3.0.CO;2-5

[40]   G. J. Zhang, S. A. Crist, A. K. McKerrow, Y. Xu, D. C. Ladehoff and W. A. See, “Autocrine IL-6 Production by Human Transitional Carcinoma Cells Upregulates Expression of the alpha5beta1 Firbonectin Receptor,” The Journal of Urology, Vol. 163, No. 5, 2000, pp. 1553-1559. doi:10.1016/S0022-5347(05)67678-1

[41]   W. Zhao, J. S. Schorey, M. Bong-Mastek, J. Ritchey, E. J. Brown and T. L. Ratliff, “Role of a Bacillus CalmetteGuerin Fibronectin Attachment Protein in BCG-Induced Antitumor Activity,” International Journal of Cancer, Vol. 86, No. 1, 2000, pp. 83-88. doi:10.1002/(SICI)1097-0215(20000401)86:1<83::AID-IJC13>3.0.CO;2-R

[42]   A. Bohle, E. van der Sloot, E. Richter, J. Gerdes, W. G. Wood and J. Gerdes, “Binding to Fibronectin (FN)—A Prerequisite Step? Investigations on the Role of FN in Intravesical BCG Immunotherapy,” Investigative Urology, Vol. 5, 1994, pp. 100-104.

[43]   R. O. Hynes and K. M. Yamada, “Fibronectins: Multifunctional Modular Glycoproteins,” The Journal of Cell Biology, Vol. 95, No. 2, 1982, pp. 369-377. doi:10.1083/jcb.95.2.369

[44]   R. O. Hynes, A. T. Destree, M. E. Perkins and D. D. Wagner, “Cell Surface Fibronectin and Oncogenic Transformation,” Journal of Supramolecular Structure, Vol. 11, No. 1, 1979, pp. 95-104. doi:10.1002/jss.400110110

[45]   H. S. Smith, J. L. Riggs and M. W. Mosesson, “Production of Fibronectin by Human Epithelial Cells in Culture,” Cancer Research, Vol. 39, No. 10, 1979, pp. 4138-4144.

[46]   N. Mutlu, L. Turkeri and K. Emerk, “Analytical and Clinical Evaluation of a New Urinary Tumor Marker: Bladder Tumor Fibronectin in Diagnosis and Follow-Up of Bladder Cancer,” Clinical Chemistry and Laboratory Medicine, Vol. 41, No. 8, 2003, pp. 1069-1074. doi:10.1515/CCLM.2003.165

[47]   E. Ioachim, M. Michael, N. E. Stavropoulos, E. Kitsiou, M. Salmas and V. Malamou-Mitsi, “A Clinicopathological Study of the Expression of Extracellular Matrix Components in Urothelial Carcinoma,” BJU International, Vol. 95, No. 4, 2005, pp. 655-659. doi:10.1111/j.1464-410X.2005.05357.x

[48]   T. Saito, Y. Tomita, T. Kawasaki, V. Bilim and K. Takahashi, “Subsequent Activation of Mitogen-Activated Protein Kinase after Adhesion of Transitional Cell Cancer Cells to Fibronectin,” Urologia Internationalis, Vol. 69, 2002, pp. 125-128. doi:10.1159/000065561

[49]   A. Bohle, A. Jurczok, P. Ardelt, T. Wulf, A. J. Ulmer, D. Jocham, et al., “Inhibition of Bladder Carcinoma Cell Adhesion by Oligopeptide Combinations in Vitro and in Vivo,” The Journal of Urology, Vol. 167, No. 1, 2002, pp. 357-363. doi:10.1016/S0022-5347(05)65468-7

[50]   V. Menendez, A. Fernandez-Suarez, J. A. Galan, M. Perez and F. Garcia-Lopez, “Diagnosis of Bladder Cancer by Analysis of Urinary Fibronectin,” Urology, Vol. 65, No. 2, 2005, pp. 284-289. doi:10.1016/j.urology.2004.09.028

[51]   M. Laufer, I. Kaver, B. Sela and H. Matzkin, “Elevated Urinary Fibronectin Levels after Transurethral Resection of Bladder Tumour: A Possible Role in Patients Failing Therapy with Bacillus Calmette-Guerin,” BJU Internation, Vol. 84, No. 4, 1999, pp. 428-432. doi:10.1046/j.1464-410x.1999.00208.x

[52]   R. E. Hurst, “Structure, Function, and pathology of Proteoglycans and Glycosaminoglycans in the Urinary Tract,” World Journal of Urology, Vol. 12, No. 1, 1994, pp. 3-10. doi:10.1007/BF00182044

[53]   K. H. Kurth, “Glycosaminoglycans and Proteoglycans in the Urinary Tract,” World Journal of Urology, Vol. 12, No. 1, 1994, p. 2.

[54]   C. L. Parsons, “The Role of the Urinary Epithelium in the Pathogenesis of Interstitial Cystitis/Prostatitis/Urethritis,” Urology, Vol. 69, No. 4, 2007, pp. 9-16. doi:10.1016/j.urology.2006.03.084

[55]   J. N'Dow, N. Jordan, C. N. Robson, D. E. Neal and J. P. Pearson, “The Bladder Does Not Appear to Have a Dynamic Secreted Continuous Mucous Gel Layer,” The Journal of Urology, Vol. 173, No. 6, 2005, pp. 2025-2031. doi:10.1097/01.ju.0000158454.47299.ae

[56]   C. L. Parsons, “A Model for the Function of Glycosaminoglycans in the Urinary Tract,” World Journal of Urology, Vol. 12, No. 1, 1994, pp. 38-42. doi:10.1007/BF00182049

[57]   V. B. Lokeshwar, M. G. Selzer, W. H. Cerwinka, M. F. Gomez, R. R. Kester, D. E. Bejany, et al., “Urinary Uronate and Sulfated Glycosaminoglycan Levels: Markers for Interstitial Cystitis Severity,” The Journal of Urology, 2005, Vol. 174, No. 1, pp. 344-349. doi:10.1097/01.ju.0000161599.69942.2e

[58]   C. L. Parsons, S. G. Mulholland and H. Anwar, “Antibacterial Activity of Bladder Surface Mucin Duplicated by Exogenous Glycosaminoglycan (Heparin),” Infection and Immunity, Vol. 24, No. 2, 1979, pp. 552-557.

 
 
Top