Dual tracer evaluation of dynamic changes in intratumoral hypoxic and proliferative states after radiotherapy of human head and neck cancer xenografts using radiolabeled FMISO and FLT

BMC Cancer

Chowdhury Nusrat Fatema¹  Songji Zhao⁴  Yan Zhao³  Wenwen Yu⁵  Ken-ichi Nishijima⁶  Koichi Yasuda²  Yoshimasa Kitagawa⁵  Nagara Tamaki³  Yuji Kuge⁶ 

[1] Department of Tracer Kinetics & Bioanalysis, Graduate School of Medicine, Hokkaido University, Kita 15, Nishi 7, Kita-ku, Sapporo 060-8638, Japan

[2] Department of Radiation Medicine, Graduate School of Medicine, Hokkaido University, Sapporo, Japan

[3] Department of Nuclear Medicine, Graduate School of Medicine, Hokkaido University, Sapporo, Japan

[4] Department of Molecular Imaging, Graduate School of Medicine, Hokkaido University, Sapporo, Japan

[5] Department of Oral Diagnosis and Medicine, Graduate School of Dental Medicine, Hokkaido University, Sapporo, Japan

[6] Central Institute of Isotope Science, Hokkaido University, Sapporo, Japan

关键词: Head and neck cancer xenograft;    Tumor reoxygenation and proliferation;    Radiotherapy;    Fluorothymidine;    Fluoromisonidazole;   

Others  :  1121049
DOI  :  10.1186/1471-2407-14-692

 received in 2014-06-19, accepted in 2014-09-17,  发布年份 2014

【 摘 要 】

Background

Radiotherapy is an important treatment strategy for head and neck cancers. Tumor hypoxia and repopulation adversely affect the radiotherapy outcome. Accordingly, fractionated radiotherapy with dose escalation or altered fractionation schedule is used to prevent hypoxia and repopulation. ¹⁸F-fluoromisonidazole (FMISO) and ¹⁸F-fluorothymidine (FLT) are noninvasive markers for assessing tumor hypoxia and proliferation, respectively. Thus, we evaluated the dynamic changes in intratumoral hypoxic and proliferative states following radiotherapy using the dual tracers of ¹⁸F-FMISO and ³H-FLT, and further verified the results by immunohistochemical staining of pimonidazole (a hypoxia marker) and Ki-67 (a proliferation marker) in human head and neck cancer xenografts (FaDu).

Methods

FaDu xenografts were established in nude mice and assigned to the non-radiation-treated control and two radiation-treated groups (10- and 20-Gy). Tumor volume was measured daily. Mice were sacrificed 6, 24, and 48 hrs and 7 days after radiotherapy. ¹⁸F-FMISO, and ³H-FLT and pimonidazole were injected intravenously 4 and 2 hrs before sacrifice, respectively. Intratumoral ¹⁸F-FMISO and ³H-FLT levels were assessed by autoradiography. Pimonidazole and Ki-67 immunohistochemistries were performed.

Results

In radiation-treated mice, tumor growth was significantly suppressed compared with the control group, but the tumor volume in these mice gradually increased with time. Visual inspection showed that intratumoral ¹⁸F-FMISO and ³H-FLT distribution patterns were markedly different. Intratumoral ¹⁸F-FMISO level did not show significant changes after radiotherapy among the non-radiation-treated control and radiation-treated groups, whereas ³H-FLT level markedly decreased to 59 and 45% of the non-radiation-treated control at 6 hrs (p < 0.0001) and then gradually increased with time in the 10- and 20-Gy-radiation-treated groups. The pimonidazole-positive hypoxic areas were visually similar in both the non-radiation-treated control and radiation-treated groups. No significant differences were observed in the percentage of pimonidazole-positive cells and Ki-67 index.

Conclusion

Intratumoral ¹⁸F-FMISO level did not change until 7 days, whereas ³H-FLT level markedly decreased at 6 hrs and then gradually increased with time after a single dose of radiotherapy. The concomitant monitoring of dynamic changes in tumor hypoxia and proliferation may provide important information for a better understanding of tumor biology after radiotherapy and for radiotherapy planning, including dose escalation and altered fractionation schedules.

【 授权许可】

   
2014 Fatema et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Argiris A, Karamouzis MV, Raben D, Ferris RL: Head and neck cancer. Lancet 2008, 371:1695-1709.
• [2]Corry J, Rischin D: Strategies to overcome accelerated repopulation and hypoxia–what have we learned from clinical trials? Semin Oncol 2004, 31:802-808.
• [3]Ljungkvist AS, Bussink J, Kaanders JH, Wiedenmann NE, Vlasman R, van der Kogel AJ: Dynamics of hypoxia, proliferation and apoptosis after irradiation in a murine tumor model. Radiat Res 2006, 165:326-336.
• [4]Isa AY, Ward TH, West CM, Slevin NJ, Homer JJ: Hypoxia in head and neck cancer. Br J Radiol 2006, 79:791-798.
• [5]Withers HR, Taylor JM, Maciejewski B: The hazard of accelerated tumor clonogen repopulation during radiotherapy. Acta Oncol 1988, 27:131-146.
• [6]Rajendran JG, Schwartz DL, O’Sullivan J, Peterson LM, Ng P, Scharnhorst J, Grierson JR, Krohn KA: Tumor hypoxia imaging with [F-18] fluoromisonidazole positron emission tomography in head and neck cancer. Clin Cancer Res 2006, 12:5435-5441.
• [7]Nagane M, Yasui H, Yamamori T, Zhao S, Kuge Y, Tamaki N, Kameya H, Nakamura H, Fujii H, Inanami O: Radiation-induced nitric oxide mitigates tumor hypoxia and radioresistance in a murine SCCVII tumor model. Biochem Biophys Res Commun 2013, 437:420-425.
• [8]Narita T, Aoyama H, Hirata K, Onodera S, Shiga T, Kobayashi H, Murata J, Terasaka S, Tanaka S, Houkin K: Reoxygenation of glioblastoma multiforme treated with fractionated radiotherapy concomitant with temozolomide: changes defined by 18F-fluoromisonidazole positron emission tomography: two case reports. Jpn J Clin Oncol 2012, 42:120-123.
• [9]Yasuda K, Onimaru R, Okamoto S, Shiga T, Katoh N, Tsuchiya K, Suzuki R, Takeuchi W, Kuge Y, Tamaki N, Shirato H: [18F] fluoromisonidazole and a new PET system with semiconductor detectors and a depth of interaction system for intensity modulated radiation therapy for nasopharyngeal cancer. Int J Radiat Oncol Biol Phys 2013, 85:142-147.
• [10]Okamoto S, Shiga T, Yasuda K, Ito YM, Magota K, Kasai K, Kuge Y, Shirato H, Tamaki N: High reproducibility of tumor hypoxia evaluated by 18F-fluoromisonidazole PET for head and neck cancer. J Nucl Med 2013, 54:201-207.
• [11]Yang YJ, Ryu JS, Kim SY, Oh SJ, Im KC, Lee H, Lee SW, Cho KJ, Cheon GJ, Moon DH: Use of 3′-deoxy-3′- [18F] fluorothymidine PET to monitor early responses to radiation therapy in murine SCCVII tumors. Eur J Nucl Med Mol Imaging 2006, 33:412-419.
• [12]Fatema CN, Zhao S, Zhao Y, Murakami M, Yu W, Nishijima K, Tamaki N, Kitagawa Y, Kuge Y: Monitoring tumor proliferative response to radiotherapy using 18F-fluorothymidine in human head and neck cancer xenograft in comparison with Ki-67. Ann Nucl Med 2013, 27:355-362.
• [13]Sugiyama M, Sakahara H, Sato K, Harada N, Fukumoto D, Kakiuchi T, Hirano T, Kohno E, Tsukada H: Evaluation of 3′-deoxy-3′-18F-fluorothymidine for monitoring tumor response to radiotherapy and photodynamic therapy in mice. J Nucl Med 2004, 45:1754-1758.
• [14]He Q, Skog S, Welander I, Tribukait B: X-irradiation effects on thymidine kinase (TK): I. TK1 and 2 in normal and malignant cells. Cell Prolif 2002, 35:69-81.
• [15]Schwartz JL, Tamura Y, Jordan R, Grierson JR, Krohn KA: Effect of p53 activation on cell growth, thymidine kinase-1 activity, and 30- deoxy-30 fluorothymidine uptake. Nucl Med Biol 2004, 31:419-423.
• [16]Tang G, Wang M, Tang X, Gan M, Luo L: Fully automated one-pot synthesis of [18F]fluoromisonidazole. Nucl Med Biol 2005, 32:553-558.
• [17]Hatano T, Zhao S, Zhao Y, Nishijima K, Kuno N, Hanzawa H, Sakamoto T, Tamaki N, Kuge Y: Biological characteristics of intratumoral [F-18] fluoromisonidazole distribution in a rodent model of glioma. Int J Oncol 2013, 42:823-830.
• [18]Murakami M, Zhao S, Zhao Y, Chowdhury NF, Yu W, Nishijima K, Takiguchi M, Tamaki N, Kuge Y: Evaluation of changes in the tumor microenvironment after sorafenib therapy by sequential histology and 18F-fluoromisonidazole hypoxia imaging in renal cell carcinoma. Int J Oncol 2012, 41:1593-1600.
• [19]Kasprzak KS, Dencker L, Larsson BS: Isotopic and nuclear analytical techniques in biological systems: a critical survey. Pure Appl Chem 1991, 63:1269-1306.
• [20]Brown RS, Leung JY, Fisher SJ, Frey KA, Ethier SP, Wahl RL: Intratumoral distribution of tritiated fluorodeoxyglucose in breast carcinoma. I. Are inflammatory cells important? J Nucl Med 1995, 36:1854-1861.
• [21]Zhao S, Kuge Y, Mochizuki T, Takahashi T, Nakada K, Sato M, Takei T, Tamaki N: Biologic correlates of intratumoral heterogeneity in 18F-FDG distribution with regional expression of glucose transporters and hexokinase-II in experimental tumor. J Nucl Med 2005, 46:675-682.
• [22]Wong CS: Experimental radiotherapy. In The Basic Science Of Oncology. Edited by Tannock IF, Hill RP. New York, NY: McGraw-Hill; 1998:322-34926.
• [23]Wither HR: Radiation biology and treatment option in radiation oncology. Cancer Res 1999, 59(Suppl):1676p-1684p.
• [24]Bussink J, Kaanders JH, Rijken PF, Raleigh JA, van der Kogel AJ: Changes in blood perfusion and hypoxia after irradiation of a human squamous cell carcinoma xenograft tumor line. Radiat Res 2000, 153:398-404.
• [25]Crokart N, Jordan BF, Baudelet C, Ansiaux R, Sonveaux P, Grégoire V, Beghein N, DeWever J, Bouzin C, Feron O, Gallez B: Early reoxygenation in tumors after irradiation: determining factors and consequences for radiotherapy regimens using daily multiple fractions. Int J Radiat Oncol Biol Phys 2005, 63:901-910.
• [26]Suit HD, Zietman A, Tomkinson K, Ramsay J, Gerweck L, Sedlacek R: Radiation response of xenografts of a human squamous cell carcinoma and a glioblastoma multiforme: a progress report. Int J Radiat Oncol Biol Phys 1990, 18:365-373.
• [27]Toulany M, Dittmann K, Krüger M, Baumann M, Rodemann HP: Radioresistance of K-Ras mutated human tumor cells is mediated through EGFR-dependent activation of PI3K-AKT pathway. Radiother Oncol 2005, 76:143-150.
• [28]Kasten-Pisula U, Menegakis A, Brammer I, Borgmann K, Mansour WY, Degenhardt S, Krause M, Schreiber A, Dahm-Daphi J, Petersen C, Dikomey E, Baumann M: The extreme radiosensitivity of the squamous cell carcinoma SKX is due to a defect in double-strand break repair. Radiother Oncol 2009, 90:257-264.
• [29]Petersen C, Eicheler W, Frömmel A, Krause M, Balschukat S, Zips D, Baumann M: Proliferation and micromilieu during fractionated irradiation of human FaDu squamous cell carcinoma in nude mice. Int J Radiat Biol 2003, 79:469-477.
• [30]Harriss W, Bezak E, Yeoh E, Hermans M: Measurement of reoxygenation during fractionated radiotherapy in head and neck squamous cell carcinoma xenografts. Australas Phys Eng Sci Med 2010, 33:251-263.
• [31]Ljungkvist AS, Bussink J, Rijken PF, Kaanders JH, van der Kogel AJ, Denekamp J: Vascular architecture, hypoxia, and proliferation in first-generation xenografts of human head-and-neck squamous cell carcinomas. Int J Radiat Oncol Biol Phys 2002, 54:215-228.