IJMPCERO  Vol.3 No.3 , August 2014
Carbon-Ion Beams Efficiently Induce Cell Killing in X-Ray Resistant Human Squamous Tongue Cancer Cells
ABSTRACT

In order to see whether carbon ion (C-ion) beams have a biological advantage over X-rays, studies were designed to examine the effects of C-ion beams on radiosensitivity in X-ray resistant cells. Clinically relevant X-ray resistant SAS-R cells derived from human tongue cancer SAS cells were used. The cells were exposed to X-rays or Spread-Out Bragg peak (SOBP) beam C-ions. Cell survival was measured using a modified high-density survival assay. Cell survival signaling and cell death signaling were analyzed using flow cytometry. The cells were labeled with putative cancer stem cell markers such as CD44 and CD326. SAS-R cells were 1.6 times more radioresistant than SAS cells after exposure to X-rays. Cell survival was similar in each cell line after exposure to C-ion beams. SAS-R cells displayed enhanced cell survival signaling when compared to SAS cells under normal conditions. On the other hand, the phosphorylation of AKT-related proteins decreased and polycaspase activities were enhanced when cells were irradiated with C-ion beams in both cell lines. More CD44 and CD326 positive cells were seen in SAS-R cells than in SAS cells. Moreover, the marker positive cell numbers significantly decreased after exposure to C-ion beams when compared to X-rays at iso-survival doses in SAS-R cells. C-ion beams efficiently induced cell killing in X-ray resistant cells which displayed activated cell survival signaling and contained more numerous cancer stem-like cells. 


Cite this paper
Takahashi, A. , Ma, H. , Nakagawa, A. , Yoshida, Y. , Kanai, T. , Ohno, T. , Kuwahara, Y. , Fukumoto, M. and Nakano, T. (2014) Carbon-Ion Beams Efficiently Induce Cell Killing in X-Ray Resistant Human Squamous Tongue Cancer Cells. International Journal of Medical Physics, Clinical Engineering and Radiation Oncology, 3, 133-142. doi: 10.4236/ijmpcero.2014.33019.
References
[1]   Joiner, M.C., van der Kogel, A.J. and Steel, G.G. (2009) Introduction: The Significance of Radiobiology and Radiotherapy for Cancer Treatment. In: Joiner, M. and van der Kogel, A., Eds., Basic Clinical Radiobiology Fourth Edition, Hodder Arnold Publication, London, 1-10.
http://dx.doi.org/10.1201/b13224-2

[2]   Kim, J.J. and Tannock, I.F. (2005) Repopulation of Cancer Cells during Therapy: An Important Cause of Treatment Failure. Nature Reviews Cancer, 5, 516-525.

[3]   Kuwahara, Y., Li, L., Baba, T., Nakagawa, H., Shimura, T., Yamamoto, Y., Ohkubo, Y. and Fukumoto, M. (2009) Clinically Relevant Radioresistant Cells Efficiently Repair DNA Double-Strand Breaks Induced by X-Rays. Cancer Science, 100, 747-752.

[4]   Kuwahara, Y., Mori, M., Oikawa, T., Shimura, T., Ohtake, Y., Mori, S., Ohkubo, Y. and Fukumoto, M. (2010) The Modified High-Density Survival Assay Is the Useful Tool to Predict the Effectiveness of Fractionated Radiation Exposure. Journal of Radiation Research, 51, 297-302.
http://dx.doi.org/10.1269/jrr.09094

[5]   Shimura, T., Kakuda, S., Ochiai, Y., Nakagawa, H., Kuwahara, Y., Takai, Y., Kobayashi, J., Komatsu, K. and Fukumoto, M. (2010) Acquired Radioresistance of Human Tumor Cells by DNA-PK/AKT/GSK3 Beta-Mediated Cyclin D1 Overexpression. Oncogene, 29, 4826-4837.

[6]   Kuwahara, Y., Oikawa, T., Ochiai, Y., Roudkenar, M.H., Fukumoto, M., Shimura, T., Ohtake, Y., Ohkubo, Y., Mori, S., Uchiyama, Y. and Fukumoto, M. (2011) Enhancement of Autophagy Is a Potential Modality for Tumors Refractory to Radiotherapy. Cell Death and Disease, 30, e177.

[7]   Shimura, T., Noma, N., Oikawa, T., Ochiai, Y., Kakuda, S., Kuwahara, Y., Takai, Y., Takahashi, A. and Fukumoto, M. (2012) Activation of the AKT/Cyclin D1/Cdk4 Survival Signaling Pathway in Radioresistant Cancer Stem Cells. Oncogenesis, 1, e12.

[8]   Shimura, T., Ochiai, Y., Noma, N., Oikawa, T., Sano, Y. and Fukumoto, M. (2013) Cyclin D1 Overexpression Perturbs DNA Replication and Induces Replication-Associated DNA Double-Strand Breaks in Acquired Radioresistant Cells. Cell Cycle, 12, 773-782.

[9]   Kuwahara, Y., Mori, M., Kitahara, S., Fukumoto, M., Ezaki, T., Mori, S., Echigo, S., Ohkubo, Y. and Fukumoto, M. (2014) Targeting of Tumor Endothelial Cells Combining 2 Gy/Day of X-Ray with Everolimus Is the Effective Modality for Overcoming Clinically Relevant Radioresistant Tumors. Cancer Medicine, 3, 310-321.

[10]   Moretti, L., Cha, Y.I., Niermann, K.J. and Lu, B. (2007) Switch between Apoptosis and Autophagy: Radiation-Induced Endoplasmic Reticulum Stress? Cell Cycle, 6, 793-798.
http://dx.doi.org/10.4161/cc.6.7.4036

[11]   Kroemer, G., Galluzzi, L., Vandenabeele, P., Abrams, J., Alnemri, E.S., Baehrecke, E.H., Blagosklonny, M.V., El-Deiry, W.S., Golstein, P., Green, D.R., Hengartner, M., Knight, R.A., Kumar, S., Lipton, S.A., Malorni, W., Nuñez, G., Peter, M.E., Tschopp, J., Yuan, J., Piacentini, M., Zhivotovsky, B. and Melino, G. (2009) Classification of Cell Death: Recommendations of the Nomenclature Committee on Cell Death 2009. Cell Death and Differentiation, 16, 3-11.

[12]   Chang, L., Graham, P.H., Hao, J., Ni, J., Bucci, J., Cozzi, P.J., Kearsley, J.H. and Li, Y. (2013) Acquisition of Epithelial-Mesenchymal Transition and Cancer Stem Cell Phenotypes Is Associated with Activation of the PI3K/ Akt/mTOR Pathway in Prostate Cancer Radioresistance. Cell Death and Disease, 4, e875.

[13]   Pardal, R., Clarke, M.F. and Morrison, S.J. (2003) Applying the Principles of Stem-Cell Biology to Cancer. Nature Reviews Cancer, 3, 895-902.

[14]   Bao, S., Wu, Q., McLendon, R.E., Hao, Y., Shi, Q., Hjelmeland, A.B., Dewhirst, M.W., Bigner, D.D. and Rich, J.N. (2006) Glioma Stem Cells Promote Radioresistance by Preferential Activation of the DNA Damage Response. Nature, 444, 756-760.

[15]   Baumann, M., Krause, M. and Hill, R. (2008) Exploring the Role of Cancer Stem Cells in Radioresistance. Nature Reviews Cancer, 8, 545-554.

[16]   Nguyen, G.H., Murph, M.M. and Chang, J.Y. (2011) Cancer Stem Cell Radioresistance and Enrichment: Where Frontline Radiation Therapy May Fail in Lung and Esophageal Cancers. Cancers, 3, 1232-1252.
http://dx.doi.org/10.3390/cancers3011232

[17]   Kanai, T., Furusawa, Y., Fukutsu, K., Itsukaichi, H., Eguchi-Kasai, K. and Ohara, H. (1997) Irradiation of Mixed Beam and Design of Spread-Out Bragg Peak for Heavy-Ion Radiotherapy. Radiation Research, 147, 78-85.
http://dx.doi.org/10.2307/3579446

[18]   Tsujii, H. and Kamada, T. (2007) A Review of Update Clinical Results of Carbon Ion Radiotherapy. Japanese Journal of Clinical Oncology, 42, 670-685.
http://dx.doi.org/10.1093/jjco/hys104

[19]   Durante, M. and Loeffler, J.S. (2010) Charged Particles in Radiation Oncology. Nature Reviews Clinical Oncology, 7, 37-43.
http://dx.doi.10.1038/nrclinonc. 2009.183

[20]   Ohno, T. (2013) Particle Radiotherapy with Carbon Ion Beams. The EPMA Journal, 4, 9.
http://dx.doi.org/10.1186/1878-5085-4-9

[21]   Nakagawa, Y., Takahashi, A., Kajihara, A., Yamakawa, N., Imai, Y., Ota, I., Okamoto, N., Mori, E., Noda, T., Furusawa, Y., Kirita, T. and Ohnishi, T. (2012) Depression of p53-Independent Akt Survival Signals in Human Oral Cancer Cells Bearing Mutated p53 Gene after Exposure to High-LET Radiation. Biochemical and Biophysical Research Communications, 423, 654-660.
http://dx.doi.org/10.1016/j.bbrc.2012.06.004

[22]   Ohno, T., Kanai, T., Yamada, S., Yusa, K., Tashiro, M., Shimada, H., Torikai, K., Yoshida, Y., Kitada, Y., Katoh, H., Ishii, T. and Nakano, T. (2011) Carbon Ion Radiotherapy at the Gunma University Heavy Ion Medical Center: New Facility Set-Up. Cancers, 3, 4046-4060.
http://dx.doi.org/10.3390/cancers3044046

[23]   Mirzayans, R., Andrais, B., Scott, A., Tessier, A. and Murray, D. (2007) A Sensitive Assay for the Evaluation of Cytotoxicity and Its Pharmacologic Modulation in Human Solid Tumor-Derived Cell Lines Exposed to Cancer-Therapeutic Agents. Journal of Pharmacy and Pharmaceutical Sciences, 10, 298s-311s.

[24]   Han, J., Fujisawa, T., Husain, S.R. and Puri, R.K. (2014) Identification and Characterization of Cancer Stem Cells in Human Head and Neck Squamous Cell Carcinoma. BMC Cancer, 14, 173.
http://dx.doi.org/10.1186/1471-2407-14-173

[25]   Waldron, N.N., Barsky, S.H., Dougherty, P.R. and Vallera, D.A. (2014) A Bispecific EpCAM/CD133-Targeted Toxin Is Effective against Carcinoma. Targeted Oncology, in press.

[26]   Ando, K. and Kase, Y. (2009) Biological Characteristics of Carbon-Ion Therapy. International Journal of Radiation Biology, 85, 715-728.
http://dx.doi.org/10.1080/09553000903072470

[27]   Suzuki, M., Kase, Y., Yamaguchi, H., Kanai, T. and Ando, K. (2000) Relative Biological Effectiveness for Cell-Killing Effect on Various Human Cell Lines Irradiated with Heavy-Ion Medical Accelerator in Chiba (HIMAC) Carbon-Ion Beams. International Journal of Radiation Oncology, Biology, Physics, 48, 241-250.
http://dx.doi.org/10.1016/S0360-3016(00)00568-X

[28]   Hirayama, R., Furusawa, Y., Fukawa, T. and Ando, K. (2005) Repair Kinetics of DNA-DSB Induced by X-Rays or Carbon Ions under Oxic and Hypoxic Conditions. Journal of Radiation Research, 46, 325-332.
http://dx.doi.org/10.1269/jrr.46.325

[29]   Takahashi, A., Yamakawa, N., Kirita, T., Omori, K., Ishioka, N., Furusawa, Y., Mori, E., Ohnishi, K. and Ohnishi, T. (2008) DNA Damage Recognition Proteins Localize along Heavy Ion Induced Tracks in the Cell Nucleus. Journal of Radiation Research, 49, 645-652.
http://dx.doi.org/10.1269/jrr.08007

[30]   Akino, Y., Teshima, T., Kihara, A., Kodera-Suzumoto, Y., Inaoka, M., Higashiyama, S., Furusawa, Y. and Matsuura, N. (2009) Carbon-Ion Beam Irradiation Effectively Suppresses Migration and Invasion of Human Non-Small-Cell Lung Cancer Cells. International Journal of Radiation Oncology, Biology, Physics, 75, 475-481.
http://dx.doi.org/10.1016/j.ijrobp.2008.12.090

[31]   Takahashi, A., Ohnishi, K., Ota, I., Asakawa, I., Tamamoto, T., Furusawa, Y., Matsumoto, H. and Ohnishi, T. (2001) p53-Dependent Thermal Enhancement of Cellular Sensitivity in Human Squamous Cell Carcinomas in Relation to LET. International Journal of Radiation Biology, 77, 1043-1051.
http://dx.doi.org/10.1080/09553000110066095

[32]   Takahashi, A., Matsumoto, H., Yuki, K., Yasumoto, J., Kajihara, A., Aoki, M., Furusawa, Y., Ohnishi, K. and Ohnishi, T. (2004) High-LET Radiation Enhanced Apoptosis But Not Necrosis Regardless of p53 Status. International Journal of Radiation Oncology, Biology, Physics, 60, 591-597.
http://dx.doi.org/10.1016/j.ijrobp.2004.05.062

[33]   Takahashi, A., Matsumoto, H., Furusawa, Y., Ohnishi, K., Ishioka, N. and Ohnishi, T. (2005) Apoptosis Induced by High-LET Radiations Is Not Affected by Cellular p53 Gene Status. International Journal of Radiation Biology, 81, 581-586.
http://dx.doi.org/10.1080/09553000500280484

[34]   Vakifahmetoglu, H., Olsson, M. and Zhivotovsky, B. (2008) Death through a Tragedy: Mitotic Catastrophe. Cell Death and Differentiation, 15, 1153-1162.
http://dx.doi.org/10.1038/cdd.2008.47

[35]   Vitale, I., Galluzzi, L., Castedo, M. and Kroemer, G. (2011) Mitotic Catastrophe: A Mechanism for Avoiding Genomic Instability. Nature Reviews Molecular Cell Biology, 12, 385-392.
http://dx.doi.org/10.1038/nrm3115

[36]   Shintani, T. and Klionsky, D.J. (2004) Autophagy in Health and Disease: A Double-Edged Sword. Science, 306, 990-995.
http://dx.doi.org/10.1126/science.1099993

[37]   Jin, X., Liu, Y., Ye, F., Liu, X., Furusawa, Y., Wu, Q., Li, F., Zheng, X., Dai, Z. and Li, Q. (2014) Role of Autophagy in High Linear Energy Transfer Radiation-Induced Cytotoxicity to Tumor Cells. Cancer Science, 105, 770-778.
http://dx.doi.org/10.1111/cas.12422

[38]   Nicholson, K.M. and Anderson, N.G. (2002) The Protein Kinase B/Akt Signalling Pathway in Human Malignancy. Cell Signaling, 14, 381-395.
http://dx.doi.org/10.1016/S0898-6568(01)00271-6

[39]   Duronio, V. (2008) The Life of a Cell: Apoptosis Regulation by the PI3K/PKB Pathway. The Biochemical Journal, 415, 333-344.
http://dx.doi.org/10.1042/BJ20081056

[40]   Cui, X., Oonishi, K., Tsujii, H., Yasuda, T., Matsumoto, Y., Furusawa, Y., Akashi, M., Kamada, T. and Okayasu, R. (2011) Effects of Carbon Ion Beam on Putative Colon Cancer Stem Cells and Its Comparison with X-Rays. Cancer Research, 71, 3676-3687.
http://dx.doi.org/10.1158/0008-5472.CAN-10-2926

[41]   Oonishi, K., Cui, X., Hirakawa, H., Fujimori, A., Kamijo, T., Yamada, S., Yokosuka, O. and Kamada, T. (2012) Different Effects of Carbon Ion Beams and X-Rays on Clonogenic Survival and DNA Repair in Human Pancreatic Cancer Stem-Like Cells. Radiotherapy and Oncology, 105, 258-265.
http://dx.doi.org/10.1016/j.radonc.2012.08.009

 
 
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