[논문]SIRT1 inhibitor에 의한 Hsp90 inhibitor의 Hsp90 샤페론 기능 억제 및 항암제 내성세포의 Hsp90 inhibitor에 대한 세포독성 증강

문현정; 이수훈; 김학봉; 이경아; 강치덕; 김선희

doi:10.5352/jls.2016.26.7.826

SIRT1 inhibitor에 의한 Hsp90 inhibitor의 Hsp90 샤페론 기능 억제 및 항암제 내성세포의 Hsp90 inhibitor에 대한 세포독성 증강
SIRT1 Inhibitor Enhances Hsp90 Inhibitor-mediated Abrogation of Hsp90 Chaperone Function and Potentiates the Cytotoxicity of Hsp90 Inhibitor in Chemo-resistant Human Cancer Cells 원문보기

생명과학회지 = Journal of life science, v.26 no.7 = no.195, 2016년, pp.826 - 834

문현정 (부산대학교 의학전문대학원 생화학교실) , 이수훈 (부산대학교 의학전문대학원 생화학교실) , 김학봉 (부산대학교 의학전문대학원 생화학교실) , 이경아 (부산대학교 의학전문대학원 생화학교실) , 강치덕 (부산대학교 의학전문대학원 생화학교실) , 김선희 (부산대학교 의학전문대학원 생화학교실)

초록
AI-Helper

본 연구는 Hsp90 inhibitor 및 SIRT1 inhibitor의 병용처리가 항암제 다제내성(MDR) 인간 암세포의 증식 억제에 효과적임을 밝혔다. SIRT1 활성 억제가 Hsp90 inhibitor인 17-AAG의 세포 독성의 효과를 증강시켰으며, 이로 인해 Hsp90 inhibitors에 대한 내성을 극복시킬 수 있음을 인간 자궁암세포인 HeyA8의 MDR 변이주인 HeyA8- MDR 세포에서 확인하였다. SIRT1 inhibitor는 Hsp90 inhibitor에 의한 Hsp90 샤페론 기능 억제를 증강시키며, ubiquitin ligase CHIP의 발현 증강을 유발하여, Hsp90 client protein 인 mutant p53 (mut p53)의 분해를 촉진시킨다. Mut p53 의 발현 감소는 암세포의 Hsp90 inhibitor 내성 획득의 가장 중요한 원인으로 지적되는 heat shock factor 1 (HSF1)/heat shock proteins (Hsps)의 발현 억제와 관련됨을 알 수 있었으며, 이는 항암제 다제내성 세포에서 SIRT1 inhibitor에 의하여 Hsp90 inhibitor에 대한 감수성이 증강되는 분자적 기전임을 밝혔다. 그러므로, SIRT1 억제에 의한 mut p53/HSF1 발현 감소가 MDR 암세포의 Hsp90 inhibitors 내성 극복에 매우 유효함을 시사하는 결과를 얻었다.

Abstract ▼ AI-Helper

The present investigation was undertaken to examine the effectiveness of the combination treatment of an Hsp90 inhibitor and a SIRT1 inhibitor on suppressing the growth of chemo-resistant human cancer cells. We showed that inhibition of SIRT1 effectively potentiated the cytotoxicity of 17-allylamino-17-demethoxygeldanamycin (17-AAG) and reversed Hsp90 inhibitor resistance in multidrug-resistant (MDR) human ovarian HeyA8-MDR cells. Amurensin G, a potent natural SIRT1 inhibitor, enhanced Hsp90 inhibitor-mediated abrogation of the Hsp90 chaperone function and accelerated degradation of mutated p53 (mut p53), an Hsp90 client protein, by up-regulation of ubiquitin ligase CHIP. Knock-down of CHIP significantly attenuated amurensin G-induced mut p53 degradation. Down-regulation of mut p53 reduced the expression of heat shock factor1 (HSF1)/heat shock proteins (Hsps), a major cause of Hsp90 inhibitor resistance, which led to sensitization of the MDR cells to the Hsp90 inhibitor by the SIRT1 inhibitor. Amurensin G potentiated cytotoxicity of the Hsp90 inhibitor in HeyA8-MDR cells through suppression of 17-AAG-induced Hsp70 and Hsp27 induction via down-regulation of mut p53/HSF1, and it caused activation of PARP and inhibition of Bcl-2. Our data suggests that SIRT1 inhibitors could be used to sensitize MDR cells to Hsp90 inhibitors, possibly through suppression of the mut p53/HSF1-dependent pathway, and a novel mut p53-directed action of SIRT1 inhibition could effectively prevent mut p53 accumulation in MDR cells.

주제어

AI 본문요약
AI-Helper

* AI 자동 식별 결과로 적합하지 않은 문장이 있을 수 있으니, 이용에 유의하시기 바랍니다.

대상 데이터

1 indicates antagonism. All experiments were carried out in triplicate.
For this study, we used HeyA8 human ovarian cancer cell line and its MDR subline HeyA8-MDR, MDR MCF-7 human breast cancer cells (MCF7-MDR) [19] and MDR human ye- logenous leukemia K562 (K562-MDR, originally named CD44^high K562) [20]. These cells were maintained in DMEM medium supplemented with 10% fetal bovine serum (FBS, GIBCO BRL, Life Technologies, Inc.

데이터처리

of at least three independent experiments. The statistical sig- nificance of differences assessed using the Student's t-test. *p<0.

이론/모형

(A) HeyA8-MDR cells were pre- treated with amurensin G (AG; 5- or 10 μg/ml) for 6 hr followed by various concentrations of 17-AAG for 96 hr. Cell survival was determined by MTT assay. Results are the means ± SEs of three experiments.

성능/효과

In this study, we showed that SIRT1 inhibition by amur- ensin G treatment effectively enhanced Hsp90 inhibitor 17-AAG cytotoxicity, and it could suppress the 17-AAG re- sistance of HeyA8-MDR cells. This result may occur because SIRT1 inhibition facilitates disruption of Hsp90 multi- chaperone complex, and up-regulation/activation of CHIP accelerates degradation/down-regulation of mut p53 and HSF1, which leads to the suppression of 17-AAG-induced Hsps and sensitization of MDR cells to 17-AAG.
Hsp90 in- hibitor induces the release and activation of HSF1, which increases transcription and translation of Hsps genes, and it leads to resistance of cancer cells to Hsp90 inhibitors [14, 24]. Our results showed that expression of Hsp70/Hsp90 in HeyA8-MDR cells was more enhanced than in HeyA8 cellsafter exposure of 17-AAG due to activation of HSF1 since the activation of HSF1 evidenced by an electrophoretic mobi- lity shift in the MDR cells. Since expression of SIRT1 as well as HSF1 also could induce MDR1/P-gp gene [29, 40], we confirmed up-regulation of SIRT1 in HeyA8-MDR cells, which might be associated with the expression of P-gp.

참고문헌 (42)

Anckar, J. and Sistonen, L. 2011. Regulation of HSF1 function in the heat stress response: implications in aging and disease. Annu. Rev. Biochem. 80, 1089-1115.

상세보기
Bagatell, R., Paine-Murrieta, G. D., Taylor, C. W., Pulcini, E. J., Akinaga, S., Benjamin, I. J. and Whitesell, L. 2000. Induction of a heat shock factor 1-dependent stress response alters the cytotoxic activity of hsp90-binding agents. Clin. Cancer Res. 6, 3312-3318.

상세보기
Bahr, O., Wick, W. and Weller, M. 2001. Modulation of MDR/MRP by wild-type and mutant p53. J. Clin. Invest. 107, 643-646.

상세보기
Banerji, U. 2009. Heat shock protein 90 as a drug target:some like it hot. Clin. Cancer Res. 15, 9-14.

상세보기
Brooks, C. L. and Gu, W. 2009. How does SIRT1 affect metabolism, senescence and cancer? Nat. Rev. Cancer 9, 123-128.

상세보기
Chin, K. V., Ueda, K., Pastan, I. and Gottesman, M. M. 1992. Modulation of activity of the promoter of the human MDR1 gene by Ras and p53. Science 255, 459-462.

상세보기
Davenport, E. L., Zeisig, A., Aronson, L. I., Moore, H. E., Hockley, S., Gonzalez, D., Smith, E. M., Powers, M. V., Sharp, S. Y., Workman, P., Morgan, G. J. and Davies, F. E. 2010. Targeting heat shock protein 72 enhances Hsp90 inhibitor-induced apoptosis in myeloma. Leukemia 24, 1804-1807.

상세보기
El-Deiry, W. S. 2003. The role of p53 in chemosensitivity and radiosensitivity. Oncogene 22, 7486-7495.

상세보기
Esser, C., Scheffner, M. and Hohfeld, J. 2005. The chaperone-associated ubiquitin ligase CHIP is able to target p53 for proteasomal degradation. J. Biol. Chem. 280, 27443-27448.

상세보기
Evans, C. G., Chang, L. and Gestwicki, J. E. 2010. Heat shock protein 70 (hsp70) as an emerging drug target. J. Med. Chem. 53, 4585-4602.

상세보기
Fan, M., Park, A. and Nephew, K. P. 2005. CHIP (carboxyl terminus of Hsc70-interacting protein) promotes basal and geldanamycin-induced degradation of estrogen receptoralpha. Mol. Endocrinol. 19, 2901-2914.

상세보기
Freed-Pastor, W. A. and Prives, C. 2012. Mutant p53: one name, many proteins. Genes Dev. 26, 1268-1286.

상세보기
Gan, L. and Mucke, L. 2008. Paths of convergence: sirtuins in aging and neurodegeneration. Neuron 58, 10-14.

상세보기
Guo, F., Rocha, K., Bali, P., Pranpat, M., Fiskus, W., Boyapalle, S., Kumaraswamy, S., Balasis, M., Greedy, B., Armitage, E. S., Lawrence, N. and Bhalla, K. 2005. Abrogation of heat shock protein 70 induction as a strategy to increase antileukemia activity of heat shock protein 90 inhibitor 17-allylamino-demethoxy geldanamycin. Cancer Res. 65, 10536-10544.

상세보기
Hong, D. S., Banerji, U., Tavana, B., George, G. C., Aaron, J. and Kurzrock, R. 2013. Targeting the molecular chaperone heat shock protein 90 (HSP90): lessons learned and future directions. Cancer Treat. Rev. 39, 375-387.

상세보기
Jhaveri, K., Taldone, T., Modi, S. and Chiosis, G. 2012. Advances in the clinical development of heat shock protein 90 (Hsp90) inhibitors in cancers. Biochim. Biophys. Acta 1823, 742-755.

상세보기
Kamal, A., Thao, L., Sensintaffar, J., Zhang, L., Boehm, M. F., Fritz, L. C. and Burrows, F. J. 2003. A high-affinity conformation of Hsp90 confers tumour selectivity on Hsp90 inhibitors. Nature 425, 407-410.

상세보기
Keshelava, N., Zuo, J. J., Chen, P., Waidyaratne, S. N., Luna, M. C., Gomer, C. J., Triche, T. J. and Reynolds, C. P. 2001. Loss of p53 function confers high-level multidrug resistance in neuroblastoma cell lines. Cancer Res. 61, 6185-6193.

상세보기
Kim, D. Y., Kim, M. J., Kim, H. B., Lee, J. W., Bae, J. H., Kim, D. W., Kang, C. D. and Kim, S. H. 2011. Suppression of multidrug resistance by treatment with TRAIL in human ovarian and breast cancer cells with high level of c-Myc. Biochim. Biophys. Acta 1812, 796-805.

상세보기
Kim, H. B., Lee, S. H., Um, J. H., Kim, M. J., Hyun, S. K., Gong, E. J., Oh, W. K., Kang, C. D. and Kim, S. H. 2015. Sensitization of chemo-resistant human chronic myeloid leukemia stem-like cells to Hsp90 inhibitor by SIRT1 inhibition. Int. J. Biol. Sci. 11, 923-934.

상세보기
Li, D., Marchenko, N. D. and Moll, U. M. 2011. SAHA shows preferential cytotoxicity in mutant p53 cancer cells by destabilizing mutant p53 through inhibition of the HDAC6-Hsp90 chaperone axis. Cell Death Differ. 18, 1904-1913.

상세보기
Li, D., Marchenko,N. D., Schulz,R., Fischer,V., Velasco-Hernandez, T., Talos, F. and Moll, U. M. 2011. Functional inactivation of endogenous MDM2 and CHIP by HSP90 causes aberrant stabilization of mutant p53 in human cancer cells. Mol. Cancer Res. 9, 577-588.

상세보기
Li, D., Yallowitz, A., Ozog, L. and Marchenko, N. 2014. A gain-of-function mutant p53-HSF1 feed forward circuit governs adaptation of cancer cells to proteotoxic stress. Cell Death Dis. 5, e1194.

상세보기
McCollum, A. K., Teneyck, C. J., Sauer, B. M., Toft, D. O. and Erlichman, C. 2006. Up-regulation of heat shock protein 27 induces resistance to 17-allylamino-demethoxygeldanamycin through a glutathione-mediated mechanism. Cancer Res. 66, 10967-10975.

상세보기
McCollum, A. K., TenEyck, C. J., Stensgard, B., Morlan, B. W., Ballman, K. V., Jenkins, R. B., Toft, D. O. and Erlichman, C. 2008. P-Glycoprotein-mediated resistance to Hsp90-directed therapy is eclipsed by the heat shock response. Cancer Res. 68, 7419-7427.

상세보기
Modi, S., Stopeck, A., Linden, H., Solit, D., Chandarlapaty, S., Rosen, N., D′Andrea, G., Dickler, M., Moynahan, M. E., Sugarman, S., Ma, W., Patil, S., Norton, L., Hannah, A. L. and Hudis, C. 2011. HSP90 inhibition is effective in breast cancer: a phase II trial of tanespimycin (17-AAG) plus trastuzumab in patients with HER2-positive metastatic breast cancer progressing on trastuzumab. Clin. Cancer Res. 17, 5132-5139.

상세보기
Moulick, K., Ahn, J. H., Zong, H., Rodina, A., Cerchietti, L., Gomes DaGama, E. M., Caldas-Lopes, E., Beebe, K., Perna, F., Hatzi, K., Vu, L. P., Zhao, X., Zatorska, D., Taldone, T., Smith-Jones, P., Alpaugh, M., Gross, S. S., Pillarsetty, N., Ku, T., Lewis, J. S., Larson, S. M., Levine, R., Erdjument-Bromage, H., Guzman, M. L., Nimer, S. D., Melnick, A., Neckers, L. and Chiosis, G. 2011. Affinity-based proteomics reveal cancer-specific networks coordinated by Hsp90. Nat. Chem. Biol. 7, 818-826.

상세보기
Muller, P., Hrstka, R., Coomber, D., Lane, D. P. and Vojtesek, B. 2008. Chaperone-dependent stabilization and degradation of p53 mutants. Oncogene 27, 3371-3383.

상세보기
Oh, W. K., Cho, K. B., Hien, T. T., Kim, T. H., Kim, H. S., Dao, T. T., Han, H. K., Kwon, S. M., Ahn, S. G., Yoon, J. H., Kim, T. H., Kim, Y. G. and Kang, K. W. 2010. Amurensin G, a potent natural SIRT1 inhibitor, rescues doxorubicin responsiveness via down-regulation of multidrug resistance 1. Mol. Pharmacol. 78, 855-864.

상세보기
Pacey, S., Gore, M., Chao, D., Banerji, U., Larkin, J., Sarker, S., Owen, K., Asad, Y., Raynaud, F., Walton, M., Judson, I., Workman, P. and Eisen, T. 2012. A Phase II trial of 17-allylamino, 17-demethoxygeldanamycin (17-AAG, tanespimycin) in patients with metastatic melanoma. Invest. New Drugs 30, 341-349.

상세보기
Piper, P. W. and Millson, S. H. 2011. Mechanisms of resistance to Hsp90 inhibitor drugs: a complex mosaic emerges. Pharmaceuticals 4, 1400-1422.

상세보기
Powers, M. V. and Workman, P. 2007. Inhibitors of the heat shock response: biology and pharmacology. FEBS Lett. 581, 3758-3769.

상세보기
Richardson, P. G., Chanan-Khan, A. A., Lonial, S., Krishnan, A. Y., Carroll, M. P., Alsina, M., Albitar, M., Berman, D., Messina, M. and Anderson, K. C. 2011. Tanespimycin and bortezomib combination treatment in patients with relapsed or relapsed and refractory multiple myeloma: results of a phase 1/2 study. Br. J. Haematol. 153, 729-740.

상세보기
Richardson, P. G., Mitsiades, C. S., Laubach, J. P., Lonial, S., Chanan-Khan, A. A. and Anderson, K. C. 2011. Inhibition of heat shock protein 90 (HSP90) as a therapeutic strategy for the treatment of myeloma and other cancers. Br. J. Haematol. 152, 367-379.

상세보기
Sampath, J., Sun, D., Kidd, V. J., Grenet, J., Gandhi, A., Shapiro, L. H., Wang, Q., Zambetti, G. P. and Schuetz, J. D. 2001. Mutant p53 cooperates with ETS and selectively up-regulates human MDR1 not MRP1. J. Biol. Chem. 276, 39359-39367.

상세보기
Solit, D. B., Osman, I., Polsky, D., Panageas, K. S., Daud, A., Goydos, J. S., Teitcher, J., Wolchok, J. D., Germino, F. J., Krown, S. E., Coit, D., Rosen, N. and Chapman, P. B. 2008. Phase II trial of 17-allylamino-17-demethoxygeldanamycin in patients with metastatic melanoma. Clin. Cancer Res. 14, 8302-8307.

상세보기
Strauss, B. E. and Haas, M. 1995. The region 3′ to the major transcriptional start site of the MDR1 downstream promoter mediates activation by a subset of mutant P53 proteins. Biochem. Biophys. Res. Commun. 217, 333-340.

상세보기
Takara, K., Sakaeda, T. and Okumura, K. 2006. An update on overcoming MDR1-mediated multidrug resistance in cancer chemotherapy. Curr. Pharm. Des. 12, 273-286.

상세보기
Taldone, T., Gozman, A., Maharaj, R. and Chiosis, G. 2008. Targeting Hsp90: small-molecule inhibitors and their clinical development. Curr. Opin. Pharmacol. 8, 370-374.

상세보기
Vilaboa, N. E., Galan, A., Troyano, A., de Blas, E. and Aller, P. 2000. Regulation of multidrug resistance 1 (MDR1)/Pglycoprotein gene expression and activity by heat-shock transcription factor 1 (HSF1). J. Biol. Chem. 275, 24970-24976.

상세보기
Westerheide, S. D., Anckar, J., Stevens, S. M., Jr., Sistonen, L. and Morimoto, R. I. 2009. Stress-inducible regulation of heat shock factor 1 by the deacetylase SIRT1. Science 323, 1063-1066.

상세보기
Yamakuchi, M. and Lowenstein, C. J. 2009. MiR-34, SIRT1 and p53: the feedback loop. Cell Cycle 8, 712-715.

상세보기

저자의 다른 논문 :

LOADING...

표제어: PCR

동의어: Packet Collision Rate

용어 설명 출처 목록 (6)

용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

내보내기 구분	파일저장 인쇄 메일전송
구성항목	기본정보 상세정보 관리번호, 논문명, 저널/프로시딩명, 저자 , 발행년, 권, 호, 시작페이지, 끝페이지, 발행기관 관리번호, 논문명, 대등논문명, 저자 , 저널/프로시딩명, 발행기관, 발행년, 발행언어, 권, 호, 시작페이지, 끝페이지, ISBN, ISSN, 주제분야, 키워드, 초록(한글), 초록(영문), 저자(소속기관)
저장형식	Text(ASCII format) Excel format RefWorks Direct Export RIS format (for Reference Manager, ProCite, EndNote), Scholar's Aids, Mendeley
메일정보	받는사람 (필수) @ 보내는사람 (선택) @ 제목 내용 KISTI 검색결과 이메일 서비스
안내	총 건의 자료가 검색되었습니다. 다운받으실 자료의 인덱스를 입력하세요. (1-10,000) 검색결과의 순서대로 최대 10,000건 까지 다운로드가 가능합니다. 데이타가 많을 경우 속도가 느려질 수 있습니다.(최대 2~3분 소요) 다운로드 파일은 UTF-8 형태로 저장됩니다. 파일의 내용이 제대로 보이지 않을실 때는 웹브라우저 상단의 보기 -> 인코딩 -> 자동선택 여부를 확인하십시오. ~ Text(ASCII format) Excel format