本研究是藉由HSPC、膽固醇和DOTMA分子組成的微脂體(liposome)包覆碳酸氫銨(ammonium bicarbonate) 飽和溶液，擬做為一新型的癌症高溫治療系統。實驗設計是利用碳酸氫銨溶液在加熱之後會分解成為二氧化碳氣體現象，先讓癌細胞吞噬碳酸氫銨微脂體，再進行加熱步驟，提高微脂體周圍環境溫度，造成微脂體內部體積膨脹，最終導致微脂體爆破，讓癌細胞受到物理性傷害而死亡。此實驗設計可以避免癌症化學性治療方法常帶來的副作用，以及癌細胞產生抗藥性的情形。碳酸氫銨分解成為二氧化碳的溫度是在36℃~60℃間，在體溫37℃下，因為碳酸氫銨分解速度較慢，二氧化碳氣體膨脹時產生的壓力不足以造成細胞死亡。然而，由MTT assay和LDH assay實驗證明，餵食細胞1mg/ml濃度的碳酸氫銨微脂體10分鐘後，在攝氏42℃(細胞溫度耐受上限)下加熱15分鐘，會導致細胞存活率下降至約50%。如果將這些經過加熱處理、已受到物理性傷害的癌細胞，再持續培養一天後，細胞會因進入寧靜狀態(quiescent state)而使其存活率更加下降至約30%。由流式細胞儀(Flow Cytometry)分析的結果發現，這些經過物理性傷害的癌細胞中，會有40%的比例停留在G0 phase，顯示此治療方法的確能減緩癌細胞的生長。另外，由細胞對微脂體胞吞機制與其在胞器內追蹤之實驗結果，可得知在經過餵食微脂體10分鐘後，再加上15分鐘的加熱時間，微脂體會經由caveolae的傳送路徑停留在early endosome與lysosome胞器。而微脂體在lysosome胞器內的破裂，會造成溶素體酵素溶解細胞膜，對細胞造成物理性傷害。因此，此治療方式是會造成癌細胞壞死(necrosis)而不是讓癌細胞進行自發性凋亡(apoptosis)。

[1] J. R. FIKE, G. T. Gobbel, T. Satoh and P. R. Stauffer, Normal brain response after interstitial microwave hyperthermia, International Journal of hyperthermia, Vol. 7, 795, 1991.
[2] J. W. Valvano, J. R. Cochran and K. R. Diller, Thermal conductivity and diffusivity of biomaterials measured with self-heating thermistors, International Journal of Themophys, Vol. 6, 301, 1985.
[3] Yukihito Ishizaka, Kazuo Maruyama, Chie Kojima, Atsushi Harada, Sadahito Aoshima, Highly temperature-sensitive liposomes based on a thermosensitive block copolymer for tumor-specific chemotherapy Biomaterials Vol. 31, 7096, 2010
[4] VanSonnenberg E, McMullen WN, Solbiati L, Livraghi T, Mueller PR, Silverman SG. Tumor ablation: principles and practice. New York: Springer SciencetBusiness Media; 2005.
[5] Z. J. Van, Heating the patient: a promising approach? , Annals of oncology, Vol. 13, 1173, 2002
[6] D. Dickson, U.K. Scientist Test Liposome Gene Therapy Technique , Nature , 365 , 4 ,1993
[7] A. B. Paul and M.G. Kevin Nebulisers for the generation of liposomal aerosols International, Journal of Pharmaceutics , 173 , 117, 1998
[8] J. Haensler, F. C. Szoka, PolyamidoamineCascade Polymers Mediate Efficient Transfection of Cells in Culture, Bioconjug. Chem. 4, 372, 1993
[9] R. R. C. New, Liposomes: a practical approach, Oxford University Press, 23, 1990
[10] B. Santanu and H. Saubhik, Interactions between cholesterol and lipids in bilayer membranes. Role of lipid headgroup and hydrocarbon chain-backbone linkage, Biochimica et Biophysica Acta , 1467 , 39 ,2000.
[11] D. L. Dan, Novel applications of liposome, TIBTECH, 16, 307, 1998.
[12] Peter Walde, Katia Cosentino, Helen Engel, and Pasquale Stano Giant Vesicles: Preparations and Applications ChemBioChem, 11, 848, 2010
[13] G. B. Stillwagon, W. E. Order, C. Guse, J. L. Klein, P. K. Leichner, S. A. Leibel, Fishman EK , M. P. Carey and T. G. Burish, Etiology and treatment of the psychological side-effects associated with cancer-chemotherapy- a critical-review and discussion, Psychological Bulletin, Vol. 104, 307, 1988.
[14] T. G. Burish and M. P. Carey, Conditioned aversive responses in cancer-chemotherapy patients - theoretical and developmental analysis, Journal of consulting and clinical psychologu, Vol. 54, 593, 1986.
[15] A. Srivastava, J.L. Maggs, D.J. Antoine, D.P. Williams, D.A. Smith, and B.K. Park Role of Reactive Metabolites in Drug-Induced Hepatotoxicity Handb Exp Pharmacol. Vol. 196, 165, 2010
[16] G. B. Stillwagon, S. E. Order, C. Guse, J. L. Klein, P. K. Leichner, S. A. Leibel and E. K. Fishman, 194 Hepatocellular cancers treated by radiation and chemotherapy combinations - toxicity and response, a radiation-therapy oncology group-study, International Journal of radiation oncology biology physics, Vol. 17, 1223, 1989.
[17] J. Seong, K. C. Keum, K. H. Han, D. Y. Lee, J. T. Lee, C. Y. Chon, Y. M. Moon, C. O. Suh and G. E. Kim, Combined transcatheter arterial chemoembolization and local radiotherapy of unresectable hepatocellular carcinoma, International Journal Of Radiation Oncology Biology Physics, Vol. 43, 393, 1999.
[18] Ezzell C, Magic bullets fly again , Scientific American, Vol. 185, 34, 2001.
[19] L. F. Fajardo, Pathological effects of hyperthermia in normal-tissues,」Cancer Research, Vol. 44, 4826. 1984.
[20] P. Sminia, J. Vanderzee, J. Wondergem and J. Haveman, A Review- Effect of hyperthermia on the central-nervous-system, International Journal of Hyperthermia, Vol. 10, 1, 1994.
[21] J. R. FIKE, G. T. Gobbel, T. Satoh and P. R. Stauffer, Normal brain response after interstitial microwave hyperthermia, International Journal of hyperthermia」, Vol. 7, 795, 1991.
[22] J. W. Valvano, J. R. Cochran and K. R. Diller, Thermal conductivity and diffusivity of biomaterials measured with self-heating thermistors,」International Journal of Themophys, Vol. 6, 301, 1985.
[23] E. Ikonen1, S. Heino and S. Lusa Caveolins and membrane cholesterol Biochem Soc Trans. Vol. 32121 ,2004
[24] P. L. Felgner, T. R. Gadek, M. Holm, R. Roman, H. W. Chan, M. Wenz, J. P. Northrop, G. M. Ringold, M. Danielson, “Lipofection - A highly efficient lipid mediated DNA Transfection Procedure” Proc. Natl. Acad. Sci. USA Vol. 84, 7413, 1987
[25] Huang SL. Ultrasound-responsive liposomes. Methods Mol Biol. Vol. 605:113 , 2010
[26] DeNardo GL, DeNardo SJ.Update: Turning the heat on cancer. Cancer Biother Radiopharm. Vol. 23:671, 2008
[27] Zahra Mamdouh, Marie-Cécile Giocondi, Raynald Laprade and Christian Le Grimellec, Temperature dependence of endocytosis in renal epithelial cells in cultureBiochimica et Biophysica Acta – Biomembranes Vol. 1282, 171, 1996
[28] Waldmann, Thomas A., Immunotherapy: past, present and future. Nature Medicine Vol.9, 269, 2003
[29] Zbigniew Darzynkiewicz, Cytometry in Cell Necrobiology: Analysis of Apoptosis and Accidental Cell Death (Necrosis). Cytometry Vol. 27, 1, 1997
[30] Ya-Ling Chiu, The characteristics, cellular uptake and intracellular trafficking of nanoparticles made of hydrophobically-modified chitosan. Journal of Controlled Release Vol. 146,152, 2010