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Constitutive Overexpression of Bcl-2, Survivin and ER Stress Chaperone GRP-78 Confers Intrinsic Radioresistance in Human Hepatocellular Carcinoma Cells: Insight into the Mechanistic Pathways Involved

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DOI: 10.4236/jct.2013.42A048    3,072 Downloads   5,417 Views   Citations


We present evidence here that abundantly expressed b-catenin-triggered NF-kB-dependent upregulation of inducible nitric oxide synthase(iNOS) found in hepatoma Mahlavu cells (RT-resistant variant designated as RR-Mal), but not in Hep 3B cells (RT-sensitive variant designated as RS-3B) is a key element contribrting to the radioresisitance through the activation of two prominent radioprotective pathways. First, high iNOS expression found in RR-Mal, but not in RS-3B cells was found to perturb calcium homeostasis that triggered ER stress response leading to the overproduction of ER chaperone GRP-78 via robust generation of cleaved ATF-6a (50 kDa) subunits and their nuclear translocation. Meanwhile, both abundantly expressed NF-κB and COX-2 found in RR-Mal cells could also provoke an increased production of PGE2 resulting in robust production of Bcl-2. Interestingly, when RR-Mal cells were treated with PDTC (a NF-κB inhibitor) or celecoxib (a COX-2 inhibitor), a concentration-dependent downregulation of Bcl-2 could be demonstrated implying that Bcl-2 overexpression was indeed mediated through NF-κB/Cox-2/PGE2 pathway. Importantly, we also unveiled that siRNA-mediated silencing of survivin in RR-Mal cells could result in a concomitant downregulation of GRP-78 due to a severe inhibition of ATF-6a (50 kDa) expression. Taken together, our data demonstrate that constitutively overexpressed b-catenin/NF-κB/iNOS and NF-κB/COX-2/PGE2 pathways that overproducing GRP-78, survivin and Bcl-2 expressions are responsible for radioresistance acquisition in RR-Mal cells. Thus, both pathways could be served as potential targets for overcoming radioresistance.

Conflicts of Interest

The authors declare no conflicts of interest.

Cite this paper

C. Wu, Y. Uen, C. Ho, Y. Tseng, T. Liu, J. Chiou and S. Leung, "Constitutive Overexpression of Bcl-2, Survivin and ER Stress Chaperone GRP-78 Confers Intrinsic Radioresistance in Human Hepatocellular Carcinoma Cells: Insight into the Mechanistic Pathways Involved," Journal of Cancer Therapy, Vol. 4 No. 2A, 2013, pp. 399-407. doi: 10.4236/jct.2013.42A048.


[1] G. M. T. Kuo and N. Savaraj, “Roles of Reactive Oxygen Species in Hepatocarcinogenesis and Drug Resistance Gene Expression in Liver Cancers,” Molecular Carcinogenesis, Vol. 45, No. 9, 2006, pp. 701-709. doi:10.1002/mc.20240
[2] D. M. Parkin, F. Bray, J. Ferlay and P. Pisani, “Global Cancer Statistics, 2002,” CA: A Cancer Journal for Clinicians, Vol. 55, No. 2, 2005, pp. 74-108. doi:55/2/74
[3] T. Zhang, H. Liu, C. Zhu, K. Briggs, Y. Kang, J. A. Fleming, and S. A. Curley, “Silencing Thioredoxin Induces Liver Cancer Cell Senescence under Hypoxia,” Hepatology Research, Vol. 42, No. 7, 2012, pp. 706-713. doi:10.1111/j.1872-034X.2012.00973.x
[4] C. C. Lin and M. C. Yin, “B Vitamins Deficiency and Decreased Anti-Oxidative State in Patients with Liver Cancer,” European Journal of Nutrition, Vol. 46, No. 5, 2007, pp. 293-299. doi:10.1007/s00394-007-0665-8
[5] B. I. Carr, “Hepatocellular Carcinoma: Current Management and Future Trends,” Gastroenterology, Vol. 127, No. 5, 2004, pp. S218-S224. doi:10.1053/j.gastro.2004.09.036
[6] M. Garcia-Barros, F. Paris, C. Cordon-Cardo, D. Lyden, S. Rafii, A. Haimovitz-Friedman, Z. Fuks and R. Kolesnick, “Tumor Response to Radiotherapy Regulated by Endothelial Cell Apoptosis,” Science, Vol. 300, No. 5622, 2003, pp. 1155-1159. doi:10.1126/science.1082504
[7] J. F. Chiou, C. J. Tai, M. T. Huang, P. L. Wei, Y. H. Wang, J. An, C. H. Wu, T. Z. Liu and Y. J. Chang, “Glucose-Regulated Protein 78 Is a Novel Contributor to Acquisition of Resistance to Sorafenib in Hepatocellular Carcinoma,” Annals of Surgical Oncology, Vol. 17, No. 2, 2010, pp. 603-612. doi:10.1245/s10434-009-0718-8
[8] C. J. Tai, H. Chin-Sheng, L. J. Kuo, P. L. Wei, H. H. Lu, H. A. Chen, T. Z. Liu, J. J. Liu, D. Z. Liu, Y. S. Ho, C. H. Wu and Y. J. Chang, “Survivin-Mediated Cancer Cell Migration through Grp78 and Epithelial-Mesenchymal Transition (Emt) Marker Expression in Mahlavu Cells,” Annals of Surgical Oncology, Vol. 19, No. 1, 2012, pp. 336-343. doi:10.1245/s10434-011-1692-5
[9] R. J. Bold, P. M. Termuhlen and D. J. McConkey, “Apoptosis, Cancer and Cancer Therapy,” Surgical Oncology, Vol. 6, No. 3, 1997, pp. 133-142. doi:S0960-7404(97)00015-7
[10] L. T. Condon, J. N. Ashman, S. R. Ell, N. D. Stafford, J. Greenman and L. Cawkwell, “Overexpression of Bcl-2 in Squamous Cell Carcinoma of the Larynx: A Marker of Radioresistance,” International Journal of Cancer, Vol. 100, No. 4, 2002, pp. 472-475. doi:10.1002/ijc.10503
[11] O. Gallo, V. Boddi, A. Calzolari, L. Simonetti, M. Trovati and S. Bianchi, “Bcl-2 Protein Expression Correlates with Recurrence and Survival in Early Stage Head and Neck Cancer Treated by Radiotherapy,” Clinical Cancer Research, Vol. 2, No. 2, 1996, pp. 261-267.
[12] P. Nix, L. Cawkwell, H. Patmore, J. Greenman and N. Stafford, “Bcl-2 Expression Predicts Radiotherapy Failure in Laryngeal Cancer,” British Journal of Cancer, Vol. 92, No. 12, 2005, pp. 2185-2189. doi:10.1038/sj.bjc.6602647
[13] E. M. Rosen, S. Fan, S. Rockwell and I. D. Goldberg, “The Molecular and Cellular Basis of Radiosensitivity: Implications for Understanding How Normal Tissues and Tumors Respond to Therapeutic Radiation,” Cancer Investigation, Vol. 17, No. 1, 1999, pp. 56-72.
[14] A. Strasser, A. W. Harris and S. Cory, “Bcl-2 Transgene Inhibits T Cell Death and Perturbs Thymic Self-Censorship,” Cell, Vol. 67, No. 5, 1991, pp. 889-899. doi:0092-8674(91)90362-3
[15] A. Kardosh, N. Soriano, P. Pyrko, Y. T. Liu, M. Jabbour, F. M. Hofman and A. H. Schonthal, “Reduced Survivin Expression and Tumor Cell Survival during Chronic Hypoxia and Further Cytotoxic Enhancement by the Cyclooxygenase-2 Inhibitor Celecoxib,” Journal of Biomedical Science, Vol. 14, No. 5, 2007, pp. 647-662. doi:10.1007/s11373-007-9173-3
[16] S. K. Sandur, A. Deorukhkar, M. K. Pandey, A. M. Pabon, S. Shentu, S. Guha, B. B. Aggarwal and S. Krishnan, “Curcumin Modulates the Radiosensitivity of Colorectal Cancer Cells by Suppressing Constitutive and Inducible Nf-Kappab Activity,” International Journal of Radiation Oncology * Biology * Physics, Vol. 75, No. 2, 2009, pp. 534-542. doi:10.1016/j.ijrobp.2009.06.034
[17] J. F. Liu, G. G. Jamieson, P. A. Drew, G. J. Zhu, S. W. Zhang, T. N. Zhu, B. E. Shan and Q. Z. Wang, “Aspirin Induces Apoptosis in Oesophageal Cancer Cells by Inhibiting the Pathway of Nf-Kappab Downstream Regulation of Cyclooxygenase-2,” ANZ Journal of Surgery, Vol. 75, No. 11, 2005, pp. 1011-1016. doi:10.1111/j.1445-2197.2005.03596.x
[18] M. Pennati, M. Folini and N. Zaffaroni, “Targeting Survivin in Cancer Therapy: Fulfilled Promises and Open Questions,” Carcinogenesis, Vol. 28, No. 6, 2007, pp. 1133-1139. doi:10.1093/carcin/bgm047
[19] K. Asanum, R. Moriai and T. Yajima, “Survivin as a Radioresistant Factor in Pancreatic Cancer,” Japanese Journal of Cancer Research, Vol. 91, No. 11, 2000, pp. 1204-1209.
[20] N. K. Sah, A. Munshi, M. Hobbs, B. Z. Carter, M. Andreeff and R. E. Meyn, “Effect of Downregulation of Survivin Expression on Radiosensitivity of Human Epidermoid Carcinoma Cells,” International Journal of Radiation Oncology * Biology * Physics, Vol. 66, No. 3, 2006, pp. 852-859. doi:10.1016/j.ijrobp.2006.06.049
[21] A. Bandino, A. Compagnone, V. Bravoco, C. Cravanzola, A. Lomartire, C. Rossetto, E. Novo, S. Cannito, L. Valfre di Bonzo, E. Zamara, R. Autelli, M. Parola and S. Colombatto, “Beta-Catenin Triggers Nuclear Factor Kappab-Dependent Up-Regulation of Hepatocyte Inducible Nitric Oxide Synthase,” The International Journal of Biochemistry & Cell Biology, Vol. 40, No. 9, 2008, pp. 1861-1871. doi:10.1016/j.biocel.2008.01.029
[22] V. Fetz, C. Bier, N. Habtemichael, R. Schuon, A. Schweitzer, M. Kunkel, K. Engels, A. F. Kovacs, S. Schneider, W. Mann, R. H. Stauber and S. K. Knauer, “Inducible No Synthase Confers Chemoresistance in Head and Neck Cancer by Modulating Survivin,” International Journal of Cancer, Vol. 124, No. 9, 2009, pp. 2033-2041. doi:10.1002/ijc.24182
[23] K. Krysan, F. H. Merchant, L. Zhu, M. Dohadwala, J. Luo, Y. Lin, N. Heuze-Vourc’h, M. Pold, D. Seligson, D. Chia, L. Goodglick, H. Wang, R. Strieter, S. Sharma and S. Dubinett, “Cox-2-Dependent Stabilization of Survivin in Non-Small Cell Lung Cancer,” The FASEB Journal, Vol. 18, No. 1, 2004, pp. 206-208. doi:10.1096/fj.03-0369fje
[24] S. W. Park, S. G. Lee, S. H. Song, D. S. Heo, B. J. Park, D. W. Lee, K. H. Kim and M. W. Sung, “The Effect of Nitric Oxide on Cyclooxygenase-2 (Cox-2) Overexpression in Head and Neck Cancer Cell Lines,” International Journal of Cancer, Vol. 107, No. 5, 2003, pp. 729-738. doi:10.1002/ijc.11498
[25] H. Yamamoto, C. Y. Ngan and M. Monden, “Cancer Cells Survive with Survivin,” Cancer Science, Vol. 99, No. 9, 2008, pp. 1709-1714. doi:10.1111/j.1349-7006.2008.00870.x
[26] M. V. Backer, J. M. Backer and P. Chinnaiyan, “Targeting the Unfolded Protein Response in Cancer Therapy,” Methods in Enzymology, Vol. 491, 2011, pp. 37-56. doi:10.1016/B978-0-12-385928-0.00003-1
[27] T. Y. Lin, J. T. Chang, H. M. Wang, S. H. Chan, C. C. Chiu, C. Y. Lin, K. H. Fan, C. T. Liao, I. H. Chen, T. Z. Liu, H. F. Li and A. J. Cheng, “Proteomics of the Radio-resistant Phenotype in Head-and-Neck Cancer: Gp96 as a Novel Prediction Marker and Sensitizing Target for Radiotherapy,” International Journal of Radiation Oncology * Biology * Physics, Vol. 78, No. 1, 2010, pp. 246-256. doi:10.1016/j.ijrobp.2010.03.002
[28] M. J. Wu, C. I. Jan, Y. G. Tsay, Y. H. Yu, C. Y. Huang, S. C. Lin, C. J. Liu, Y. S. Chen, J. F. Lo and C. C. Yu, “Elimination of Head and Neck Cancer Initiating Cells through Targeting Glucose Regulated Protein78 Signaling,” Molecular Cancer, Vol. 9, 2010, p. 283. doi:10.1186/1476-4598-9-283
[29] Y. J. Chang, C. J. Tai, L. J. Kuo, P. L. Wei, H. H. Liang, T. Z. Liu, W. Wang, Y. S. Ho, C. H. Wu and M. T. Huang, “Glucose-Regulated Protein 78 (Grp78) Mediated the Efficacy to Curcumin Treatment on Hepatocellular Carcinoma,” Annals of Surgical Oncology, Vol. 18, No. 8, 2011, pp. 2395-2403. doi:10.1245/s10434-011-1597-3
[30] S. P. Ermakova, B. S. Kang, B. Y. Choi, H. S. Choi, T. F. Schuster, W. Y. Ma, A. M. Bode and Z. Dong, “(-)-Epigallocatechin Gallate Overcomes Resistance to Etoposide-Induced Cell Death by Targeting the Molecular Chaperone Glucose-Regulated Protein 78,” Cancer Research, Vol. 66, No. 18, 2006, pp. 9260-9269. doi:10.1158/0008-5472.CAN-06-1586

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