進階搜尋


   電子論文尚未授權公開,紙本請查館藏目錄
(※如查詢不到或館藏狀況顯示「閉架不公開」,表示該本論文不在書庫,無法取用。)
系統識別號 U0026-1807201916444200
論文名稱(中文) 拉帕替尼(lapatinib)與索拉非尼(sorafenib) 對於心臟細胞上不同離子電流的區別性抑制作用
論文名稱(英文) Differential inhibitory actions by lapatinib or sorafenib, known as multi-targeted tyrosine kinase inhibitors, on different types of ionic currents in heart cells
校院名稱 成功大學
系所名稱(中) 生理學研究所
系所名稱(英) Department of Physiology
學年度 107
學期 2
出版年 108
研究生(中文) 李凱森
研究生(英文) Kai-Sen Lee
學號 S36061072
學位類別 碩士
語文別 英文
論文頁數 48頁
口試委員 指導教授-吳勝男
口試委員-陳珮君
口試委員-劉彥青
口試委員-陸德齡
中文關鍵字 酪胺酸激酶抑制劑  鉀離子電流  鈉離子電流  動作電位持續時間  心臟細胞 
英文關鍵字 Tyrosine kinase inhibitor  K+ currents  Na+ current  action potential duration  heart cell 
學科別分類
中文摘要 拉帕替尼(lapatinib)與索拉非尼(sorafenib)是一種酪胺酸激酶抑制劑。透過其抑制作用影響細胞下游信號,抑制腫瘤細胞的生長和血管新生,近來被核准使用於癌症治療。然而此類藥物存在著副作用,其中一項為心律不整。在先前的研究指出,有些患者服用此類酪胺酸激酶抑制劑後心電圖中出現QT間隔延長的現象。QT延長有很高機率會導致tosade de pointes這個多型性心室心搏過速,可能會導致暈眩甚至是猝死。在過去的研究當中,緩慢激活延遲修正的鉀離子電流(slowly activating delayed-rectifier K+ current)與erg調控的鉀離子電流(erg-mediated K+ current)的表現能夠影響心電圖QT間隔的時間長短。然而此類酪胺酸激酶抑制劑對心臟細胞膜上是否產生影響目前還是未知的。在本篇研究中我們利用細胞膜箝制的技術(patch-clamp),評估拉帕替尼與索拉非尼這類酪胺酸激酶抑制劑對於心臟細胞(H9c2和neonatal rat ventricular cardiomyocytes)上離子電流的影響。包含緩慢激活延遲修正的鉀離子電流、erg調控的鉀離子電流、向內修正鉀離子電流(inwardly rectifying K+ current)和電壓門控的鈉離子電流(voltage-gated Na+ current)。研究結果顯示,在給予藥物後緩慢激活延遲修正的鉀離子電流的振幅會受到抑制,此電流動力學也會受到影響。給予藥物後也會讓此電流的活化曲線朝向更去極化電位移動,而曲線的門控電荷並沒有顯著變化。另外也發現此類藥物也會對erg調控的鉀離子電流、向內修正鉀離子電流和電壓門控的鈉離子電流有不同程度的抑制作用。在給予這類藥物後,心臟細胞的動作電位持續時間也會延長。綜合實驗的結果,此類藥物對於心臟細胞中離子通道具有臨床相關性,且可能有直接作用。
英文摘要 Lapatinib (LAP) and sorafenib (SOR) are multi-targeted, small molecules which belong to a family of tyrosine kinase inhibitors (TKIs). They have been recently approved for the treatment of a variety of malignant cancers. They are recognized to influence the signaling pathway of tumor cell proliferation, and to inhibit cell surface kinases, thus decreasing the angiogenesis. However, some TKIs have been notably reported to cause a significant prolongation of electrocardiographic QTc interval. The prolongation of QTc intervals may cause the tosade de pointes tachyarrhythmia, which may result in syncope or sudden death. Previous studies have shown that changes in the amplitude and kinetics of the slowly activating delayed-rectifier K+ current IK(S) and erg-mediated K+ current IK(erg) can potentially alter the QT interval duration. Therefore, in this study, we wanted to evaluate the possible effects of LAP or SOR on ionic currents including IK(S), IK(erg), inwardly rectifying K+ current IK(IR), voltage-gated Na+ current (INa) in heart-derived H9C2 cells and in neonatal rat ventricular cardiomyocytes by using the patch-clamp technique. Findings from these results have shown that the presence of LAP or SOR was able to suppress IK(S) amplitude as well as to alter the IK(S) activation and deactivation time courses. The presence of LAP can also shift the activation curve of IK(S) towards more depolarized potentials with no significant change in gating charge of the curve. Therefore, the presence of LAP or SOR can directly suppress the amplitudes of IK(S), IK(erg) , IK(IR) and INa with different potency observed in heart cells. Addition of LAP or SOR can also cause the prolongation of action potential duration. The targets for these ion channels in heart cells tend to be direct and could be of clinical relevance for this group of drugs, if similar findings occur in vivo.
論文目次 Table of Contents I
Figure Contents II
中文摘要 1
Abstract 2
Acknowledgement 4
Introduction 5
Materials and Methods 8
Chemicals and solutions 8
Cell preparations 9
Isolation and culture of neonatal rat ventricular myocytes 10
Electrophysiological measurements 10
Data analyses 12
Statistical analysis 12
Kinetic study of LAP or SOR effect on the slow component of IK(S) deactivation time course 13
Results 15
Effect of high K+ solution on K+ current in H9c2 cells 15
Concentration-dependent effects of LAP or SOR on IK(S) in H9c2 cells 16
Effect of LAP on the I-V relationship of IK(S) in H9c2 cells 18
Effect of LAP and LAP plus VEGF on IK(S) in H9c2 cells. 19
Inhibitory effect of LAP on IK(S) elicited by a train of rapid repetitive depolarizations 19
Effect of LAP in erg-mediated K+ current (IK(erg)) in H9c2 cells 20
Suppressive effect of LAP and SOR on IK(IR) amplitude measured from cultured neonatal rat ventricular myocytes 21
Effect of LAP on voltage-gated Na+ current (INa) in cultured neonatal rat ventricular myocytes 21
Effect of LAP on membrane potential recorded from cultured neonatal rat ventricular myocytes 22
Discussion 24
References 28
Figures 35
Figure Legends 44

參考文獻 Adderley, S.P., E.A. Dufaux, M. Sridharan, E.A. Bowles, M.S. Hanson, A.H. Stephenson, M.L. Ellsworth, and R.S. Sprague. 2009. Iloprost- and isoproterenol-induced increases in cAMP are regulated by different phosphodiesterases in erythrocytes of both rabbits and humans. American journal of physiology. Heart and circulatory physiology. 296:H1617-1624.
Calistri, L., C. Cordopatri, C. Nardi, E. Gianni, F. Marra, and S. Colagrande. 2017. Sudden cardiac death in a patient with advanced hepatocellular carcinoma with good response to sorafenib treatment: A case report with literature analysis. Molecular and clinical oncology. 6:389-396.
Chaar, M., J. Kamta, and S. Ait-Oudhia. 2018. Mechanisms, monitoring, and management of tyrosine kinase inhibitors-associated cardiovascular toxicities. OncoTargets and therapy. 11:6227-6237.
Chang, W.T., and S.N. Wu. 2018. Activation of voltage-gated sodium current and inhibition of erg-mediated potassium current caused by telmisartan, an antagonist of angiotensin II type-1 receptor, in HL-1 atrial cardiomyocytes. Clinical and experimental pharmacology & physiology. 45:797-807.
Chien, A.J., P.N. Munster, M.E. Melisko, H.S. Rugo, J.W. Park, A. Goga, G. Auerback, E. Khanafshar, K. Ordovas, K.M. Koch, and M.M. Moasser. 2014. Phase I dose-escalation study of 5-day intermittent oral lapatinib therapy in patients with human epidermal growth factor receptor 2-overexpressing breast cancer. Journal of clinical oncology : official journal of the American Society of Clinical Oncology. 32:1472-1479.
Ding, W.G., F. Toyoda, and H. Matsuura. 2002. Blocking action of chromanol 293B on the slow component of delayed rectifier K+ current in guinea‐pig sino‐atrial node cells. British journal of pharmacology. 137:253-262.
Fredj, S., N. Lindegger, K.J. Sampson, P. Carmeliet, and R.S. Kass. 2006. Altered Na+ channels promote pause-induced spontaneous diastolic activity in long QT syndrome type 3 myocytes. Circulation research. 99:1225-1232.
Giles, W., T. Nakajima, K. Ono, and E.F. Shibata. 1989. Modulation of the delayed rectifier K+ current by isoprenaline in bull-frog atrial myocytes. The Journal of physiology. 415:233-249.
Groenendijk, F.H., W.W. Mellema, E. van der Burg, E. Schut, M. Hauptmann, H.M. Horlings, S.M. Willems, M.M. van den Heuvel, J. Jonkers, E.F. Smit, and R. Bernards. 2015. Sorafenib synergizes with metformin in NSCLC through AMPK pathway activation. International journal of cancer. 136:1434-1444.
Hoff, P.M., R.A. Wolff, K. Bogaard, S. Waldrum, and J.L. Abbruzzese. 2006. A Phase I study of escalating doses of the tyrosine kinase inhibitor semaxanib (SU5416) in combination with irinotecan in patients with advanced colorectal carcinoma. Japanese journal of clinical oncology. 36:100-103.
Kimura, T., M. Uesugi, K. Takase, N. Miyamoto, and K. Sawada. 2017. Hsp90 inhibitor geldanamycin attenuates the cytotoxicity of sunitinib in cardiomyocytes via inhibition of the autophagy pathway. Toxicology and applied pharmacology. 329:282-292.
Kloth, J.S., A. Pagani, M.C. Verboom, A. Malovini, C. Napolitano, W.H. Kruit, S. Sleijfer, N. Steeghs, A. Zambelli, and R.H. Mathijssen. 2015. Incidence and relevance of QTc-interval prolongation caused by tyrosine kinase inhibitors. British journal of cancer. 112:1011-1016.
Kuo, P.C., C.J. Yang, Y.C. Lee, P.C. Chen, Y.C. Liu, and S.N. Wu. 2018. The comprehensive electrophysiological study of curcuminoids on delayed-rectifier K(+) currents in insulin-secreting cells. European journal of pharmacology. 819:233-241.
Liu, Y.C., Y.J. Wang, and S.N. Wu. 2008. The mechanisms of propofol-induced block on ion currents in differentiated H9c2 cardiac cells. European journal of pharmacology. 590:93-98.
Ma, D., H. Wei, J. Lu, D. Huang, Z. Liu, L.J. Loh, O. Islam, R. Liew, W. Shim, and S.A. Cook. 2015. Characterization of a novel KCNQ1 mutation for type 1 long QT syndrome and assessment of the therapeutic potential of a novel IKs activator using patient-specific induced pluripotent stem cell-derived cardiomyocytes. Stem cell research & therapy. 6:39.
Molife, L.R., E.J. Dean, M. Blanco-Codesido, M.G. Krebs, A.T. Brunetto, A.P. Greystoke, G. Daniele, L. Lee, G. Kuznetsov, K.T. Myint, K. Wood, B. de Las Heras, and M.R. Ranson. 2014. A phase I, dose-escalation study of the multitargeted receptor tyrosine kinase inhibitor, golvatinib, in patients with advanced solid tumors. Clinical cancer research : an official journal of the American Association for Cancer Research. 20:6284-6294.
Nakajima, T., S. Wu, H. Irisawa, and W. Giles. 1990. Mechanism of acetylcholine-induced inhibition of Ca current in bullfrog atrial myocytes. The Journal of general physiology. 96:865-885.
Paech, F., J. Bouitbir, and S. Krahenbuhl. 2017. Hepatocellular Toxicity Associated with Tyrosine Kinase Inhibitors: Mitochondrial Damage and Inhibition of Glycolysis. Frontiers in pharmacology. 8:367.
Perry, M., F.B. Sachse, J. Abbruzzese, and M.C. Sanguinetti. 2009. PD-118057 contacts the pore helix of hERG1 channels to attenuate inactivation and enhance K+ conductance. Proceedings of the National Academy of Sciences of the United States of America. 106:20075-20080.
Reyes, R., N.A. Wani, K. Ghoshal, S.T. Jacob, and T. Motiwala. 2017. Sorafenib and 2-Deoxyglucose Synergistically Inhibit Proliferation of Both Sorafenib-Sensitive and -Resistant HCC Cells by Inhibiting ATP Production. Gene expression. 17:129-140.
Robbins, J. 2001. KCNQ potassium channels: physiology, pathophysiology, and pharmacology. Pharmacology & therapeutics. 90:1-19.
Sharma, A., P.W. Burridge, W.L. McKeithan, R. Serrano, P. Shukla, N. Sayed, J.M. Churko, T. Kitani, H. Wu, A. Holmstrom, E. Matsa, Y. Zhang, A. Kumar, A.C. Fan, J.C. Del Alamo, S.M. Wu, J.J. Moslehi, M. Mercola, and J.C. Wu. 2017. High-throughput screening of tyrosine kinase inhibitor cardiotoxicity with human induced pluripotent stem cells. Science translational medicine. 9.
Shell, S.A., L. Lyass, P.B. Trusk, K.J. Pry, R.L. Wappel, and S.S. Bacus. 2008. Activation of AMPK is necessary for killing cancer cells and sparing cardiac cells. Cell cycle (Georgetown, Tex.). 7:1769-1775.
Shitara, K., T.M. Kim, T. Yokota, M. Goto, T. Satoh, J.H. Ahn, H.S. Kim, S. Assadourian, C. Gomez, M. Harnois, S. Hamauchi, T. Kudo, T. Doi, and Y.J. Bang. 2017. Phase I dose-escalation study of the c-Met tyrosine kinase inhibitor SAR125844 in Asian patients with advanced solid tumors, including patients with MET-amplified gastric cancer. Oncotarget. 8:79546-79555.
Smith, J.L., C.L. Anderson, D.E. Burgess, C.S. Elayi, C.T. January, and B.P. Delisle. 2016. Molecular pathogenesis of long QT syndrome type 2. Journal of arrhythmia. 32:373-380.
So, E.C., C.H. Hsing, C.H. Liang, and S.N. Wu. 2012. The actions of mdivi-1, an inhibitor of mitochondrial fission, on rapidly activating delayed-rectifier K(+) current and membrane potential in HL-1 murine atrial cardiomyocytes. European journal of pharmacology. 683:1-9.
So, E.C., K.C. Wu, F.C. Kao, and S.N. Wu. 2014. Effects of midazolam on ion currents and membrane potential in differentiated motor neuron-like NSC-34 and NG108-15 cells. European journal of pharmacology. 724:152-160.
So, E.C., S.N. Wu, P.C. Wu, H.Z. Chen, and C.J. Yang. 2017. Synergistic Inhibition of Delayed Rectifier K+ and Voltage-Gated Na+ Currents by Artemisinin in Pituitary Tumor (GH3) Cells. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology. 41:2053-2066.
Spears, D.A., and M.H. Gollob. 2015. Genetics of inherited primary arrhythmia disorders. The application of clinical genetics. 8:215-233.
Sung, R.J., C.-T. Kuo, S.-N. Wu, W.-T. Lai, N. Luqman, and N.-Y. Chan. 2008. Sudden cardiac death syndrome: age, gender, ethnicity, and genetics. Acta Cardiologica Sinica. 24:65-74.
Sung, R.J., S.N. Wu, J.S. Wu, H.D. Chang, and C.H. Luo. 2006. Electrophysiological mechanisms of ventricular arrhythmias in relation to Andersen-Tawil syndrome under conditions of reduced IK1: a simulation study. American journal of physiology. Heart and circulatory physiology. 291:H2597-2605.
Taylor, K.C., and C.R. Sanders. 2017. Regulation of KCNQ/Kv7 family voltage-gated K(+) channels by lipids. Biochimica et biophysica acta. Biomembranes. 1859:586-597.
Testai, L., V. Barrese, M.V. Soldovieri, P. Ambrosino, A. Martelli, I. Vinciguerra, F. Miceli, I.A. Greenwood, M.J. Curtis, M.C. Breschi, M.J. Sisalli, A. Scorziello, M.J. Canduela, P. Grandes, V. Calderone, and M. Taglialatela. 2016. Expression and function of Kv7.4 channels in rat cardiac mitochondria: possible targets for cardioprotection. Cardiovascular research. 110:40-50.
Thompson, E., J. Eldstrom, M. Westhoff, D. McAfee, E. Balse, and D. Fedida. 2017. cAMP-dependent regulation of IKs single-channel kinetics. The Journal of general physiology. 149:781-798.
Wang, Y.J., B.S. Chen, M.W. Lin, A.A. Lin, H. Peng, R.J. Sung, and S.N. Wu. 2008. Time-dependent block of ultrarapid-delayed rectifier K+ currents by aconitine, a potent cardiotoxin, in heart-derived H9c2 myoblasts and in neonatal rat ventricular myocytes. Toxicological sciences : an official journal of the Society of Toxicology. 106:454-463.
Wu, S.N., H.D. Chang, and R.J. Sung. 2006. Cocaine-induced inhibition of ATP-sensitive K+ channels in rat ventricular myocytes and in heart-derived H9c2 cells. Basic & clinical pharmacology & toxicology. 98:510-517.
Wu, S.N., B.S. Chen, and Y.C. Lo. 2011. Evidence for aconitine-induced inhibition of delayed rectifier K(+) current in Jurkat T-lymphocytes. Toxicology. 289:11-18.
Wu, S.N., H.Z. Chen, Y.H. Chou, Y.M. Huang, and Y.C. Lo. 2015. Inhibitory actions by ibandronate sodium, a nitrogen-containing bisphosphonate, on calcium-activated potassium channels in Madin-Darby canine kidney cells. Toxicology reports. 2:1182-1193.
Wu, S.N., J.H. Chern, S. Shen, H.H. Chen, Y.T. Hsu, C.C. Lee, M.H. Chan, M.C. Lai, and F.S. Shie. 2017. Stimulatory actions of a novel thiourea derivative on large-conductance, calcium-activated potassium channels. Journal of cellular physiology. 232:3409-3421.
Wu, S.N., H.F. Li, C.R. Jan, and A.Y. Shen. 1999. Inhibition of Ca2+-activated K+ current by clotrimazole in rat anterior pituitary GH3 cells. Neuropharmacology. 38:979-989.
Wu, S.N., A.Z. Wu, and R.J. Sung. 2007. Identification of two types of ATP-sensitive K+ channels in rat ventricular myocytes. Life sciences. 80:378-387.
Anumonwo, J.M., and A.N. Lopatin. 2010. Cardiac strong inward rectifier potassium channels. Journal of molecular and cellular cardiology. 48:45-54.
Marban, E., T. Yamagishi, and G.F. Tomaselli. 1998. Structure and function of voltage‐gated sodium channels. The Journal of physiology. 508:647-657.
Martinson, A.S., D.B. van Rossum, F.H. Diatta, M.J. Layden, S.A. Rhodes, M.Q. Martindale, and T. Jegla. 2014. Functional evolution of Erg potassium channel gating reveals an ancient origin for IKr. Proceedings of the National Academy of Sciences of the United States of America. 111:5712-5717.
Osteen, J.D., K.J. Sampson, and R.S. Kass. 2010. The cardiac IKs channel, complex indeed. Proceedings of the National Academy of Sciences of the United States of America. 107:18751-18752.
Wang, J.J., and Y. Li. 2016. KCNQ potassium channels in sensory system and neural circuits. Acta pharmacologica Sinica. 37:25-33.
論文全文使用權限
  • 同意授權校內瀏覽/列印電子全文服務,於2020-03-01起公開。
  • 同意授權校外瀏覽/列印電子全文服務,於2020-03-01起公開。


  • 如您有疑問,請聯絡圖書館
    聯絡電話:(06)2757575#65773
    聯絡E-mail:etds@email.ncku.edu.tw