백혈병세포에서 PTEN 발현에 대한 Ciglitazone과 retinoic Acid의 항진 작용 Ciglitazone, in Combination with All trans Retinoic Acid, Synergistically Induces PTEN Expression in HL-60 Cells원문보기
Peroxisome proliferator-activated receptor-gamma$(PPAR{\gamma})$ 는 DNA와 결합하기 위해 retinoid-X receptor(RXR)와 heterodimer를 형성해야만 한다. 그리고 전사에 대한 최대활성은 수용체에 대한 리간드 특이성에 의하는 것으로 생각되고 있다. 활성화된 $(PPAR{\gamma})$ 와 $(PPAR{\gamma})$ 리간드는 종양억제 PTEN의 조절을 통해 종양세포의 성장에 영향을 끼치게 된다. 본 연구의 목적은 $(PPAR{\gamma})$ ligand, ciglitazone그리고 RXR ligand로 동시에 자극하였을 때 급성전골수성백혈병(APL) 세포에 대해 이들이 함께 PTEN upregulate를 조절할 수 있는지를 결정하기 위함이다. 그리고 이들 세포의 성장과 분화주기에 대해 강력한 억제 능이 있는지를 결정하고자 하였다. 즉, 사람의 백혈병세포주인 HL-60세포에 all-trans-retinol과 ciglutazone을 노출시킨 뒤 PTEN 발현에 대한 측정을 위해 RT-PCR법으로 PTEN mRNA 발현 정도를 확인하고 western blot으로 분석하였다 세포주기의 분석은 propidium iodide(PI) 염색법과 FACScan으로 분석하였고, HL-60 cells에서 $(PPAR{\gamma})$ ligand, ciglitazone, 그리고 RXR ligand, retinoic acid 그리고 upregulated PTEN 발현에 대한 time- and dose-dependent방법으로 각각 확인하였던 바 ciglitazone과 retinoic acid를 동시 조합하여 처치하였을 때 유의적인 효과를 인정할 수 있었다. 더욱이 이들 혼합 물질은 세포의 성장과 G, phase를 동시 억제하는 능력이 있었다. 그러므로 $(PPAR{\gamma})$의 활성에 있어 RXR heterodimer가 사람의 백혈병세포에 대한 조절 경로로서 존재하며, PTEN의 upregulation을 통해 백혈병을 조절하기 때문에 백혈병의 예방 및 치료 접근에 $(PPAR{\gamma})$와 RXR ligands가 중요한 역할을 할 것이다.
Peroxisome proliferator-activated receptor-gamma$(PPAR{\gamma})$ 는 DNA와 결합하기 위해 retinoid-X receptor(RXR)와 heterodimer를 형성해야만 한다. 그리고 전사에 대한 최대활성은 수용체에 대한 리간드 특이성에 의하는 것으로 생각되고 있다. 활성화된 $(PPAR{\gamma})$ 와 $(PPAR{\gamma})$ 리간드는 종양억제 PTEN의 조절을 통해 종양세포의 성장에 영향을 끼치게 된다. 본 연구의 목적은 $(PPAR{\gamma})$ ligand, ciglitazone그리고 RXR ligand로 동시에 자극하였을 때 급성전골수성백혈병(APL) 세포에 대해 이들이 함께 PTEN upregulate를 조절할 수 있는지를 결정하기 위함이다. 그리고 이들 세포의 성장과 분화주기에 대해 강력한 억제 능이 있는지를 결정하고자 하였다. 즉, 사람의 백혈병세포주인 HL-60세포에 all-trans-retinol과 ciglutazone을 노출시킨 뒤 PTEN 발현에 대한 측정을 위해 RT-PCR법으로 PTEN mRNA 발현 정도를 확인하고 western blot으로 분석하였다 세포주기의 분석은 propidium iodide(PI) 염색법과 FACScan으로 분석하였고, HL-60 cells에서 $(PPAR{\gamma})$ ligand, ciglitazone, 그리고 RXR ligand, retinoic acid 그리고 upregulated PTEN 발현에 대한 time- and dose-dependent방법으로 각각 확인하였던 바 ciglitazone과 retinoic acid를 동시 조합하여 처치하였을 때 유의적인 효과를 인정할 수 있었다. 더욱이 이들 혼합 물질은 세포의 성장과 G, phase를 동시 억제하는 능력이 있었다. 그러므로 $(PPAR{\gamma})$의 활성에 있어 RXR heterodimer가 사람의 백혈병세포에 대한 조절 경로로서 존재하며, PTEN의 upregulation을 통해 백혈병을 조절하기 때문에 백혈병의 예방 및 치료 접근에 $(PPAR{\gamma})$와 RXR ligands가 중요한 역할을 할 것이다.
Peroxisome proliferatorr-activated receptor-gamma $(PPAR{\gamma})$ must form a heterodimer with the retinoid-X receptor (RXR) to bind DNA, and its transcriptional activity is thought to be maximized by ligands specific for either receptor. Activated $(PPAR{\gamma})$ and $(...
Peroxisome proliferatorr-activated receptor-gamma $(PPAR{\gamma})$ must form a heterodimer with the retinoid-X receptor (RXR) to bind DNA, and its transcriptional activity is thought to be maximized by ligands specific for either receptor. Activated $(PPAR{\gamma})$ and $(PPAR{\gamma})$ ligands may influence tumor growth through regulation of the tumor suppressor PTEN. Our aim in this study was to determine whether co-stimulation with the $(PPAR{\gamma})$ ligand, ciglitazone, and RXR ligand can synergistically upregulate PTEN in human acute promyelocytic leukemia (APL) cells and consequently potentate the inhibition of cell growth and cell cycle progression of these cells. Human leukemia cell line, HL-60 cells were exposed to all-trans-retinol and ciglutazone. The PTEN expression was measured as the level of PTEN mRNA expression by RT-PCR and as the level of PTEN expression by western blot analysis. Cell cycle analysis was carried out by a propidium iodide (PI) staining method and analyzed with a FACScan. The $(PPAR{\gamma})$ ligand, ciglitazone, and the RXR ligand, retinoic acid, upregulated PTEN expression by HL-60 cells in time- and dose-dependent manners, respectively. This was significantly enhanced by a combination of both ciglitazone and retinoic acid. Moreover, these compounds synergistically induced arrests of both cell growth and the $G_l$ phase of the cell cycle. Thus, the activation of the $(PPAR{\gamma})$:RXR heterodimer may represent a regulatory pathway for human leukemia cells and there may be important roles for $(PPAR{\gamma})$ and RXR ligands in prophylactic and therapeutic approaches fur controlling leukemia through the upregulation of PTEN.
Peroxisome proliferatorr-activated receptor-gamma $(PPAR{\gamma})$ must form a heterodimer with the retinoid-X receptor (RXR) to bind DNA, and its transcriptional activity is thought to be maximized by ligands specific for either receptor. Activated $(PPAR{\gamma})$ and $(PPAR{\gamma})$ ligands may influence tumor growth through regulation of the tumor suppressor PTEN. Our aim in this study was to determine whether co-stimulation with the $(PPAR{\gamma})$ ligand, ciglitazone, and RXR ligand can synergistically upregulate PTEN in human acute promyelocytic leukemia (APL) cells and consequently potentate the inhibition of cell growth and cell cycle progression of these cells. Human leukemia cell line, HL-60 cells were exposed to all-trans-retinol and ciglutazone. The PTEN expression was measured as the level of PTEN mRNA expression by RT-PCR and as the level of PTEN expression by western blot analysis. Cell cycle analysis was carried out by a propidium iodide (PI) staining method and analyzed with a FACScan. The $(PPAR{\gamma})$ ligand, ciglitazone, and the RXR ligand, retinoic acid, upregulated PTEN expression by HL-60 cells in time- and dose-dependent manners, respectively. This was significantly enhanced by a combination of both ciglitazone and retinoic acid. Moreover, these compounds synergistically induced arrests of both cell growth and the $G_l$ phase of the cell cycle. Thus, the activation of the $(PPAR{\gamma})$:RXR heterodimer may represent a regulatory pathway for human leukemia cells and there may be important roles for $(PPAR{\gamma})$ and RXR ligands in prophylactic and therapeutic approaches fur controlling leukemia through the upregulation of PTEN.
* AI 자동 식별 결과로 적합하지 않은 문장이 있을 수 있으니, 이용에 유의하시기 바랍니다.
제안 방법
Our aim in this study was to determine whether costimulation with the PPARy ligand, ciglitazone, and RXR ligand can synergistically upregulate PTEN in human APL cells and consequently potentiate the inhibition of cell growth and cell cycle progression of these cells.
대상 데이터
A human leukemia cell line (HL-60) was obtained from the American Type Culture Collection (Rockville, MD, USA). The cells were cultured in RPMI 1640 medium containing 10% fetal calf serum, 2 mM glutamine, antibiotics (Penicillin G 60 mg/L, Streptomycin 100 mg/L, Amphotericin B 50 |il/L) under a humid atmosphere (5% CCe, 95% air).
데이터처리
All experimental data are presented as means 土 standard deviations (SD). Statistical analyses were performed using ANOVA and Student s t tests, and P < 0.05 was considered to be significant.
이론/모형
The cell proliferation assay was performed using the Quick Cell Proliferation Assay Kit (BioVison, Mountain View, CA, USA), which is based on the cleavage of the tetrazolium salt to formazan by cellular mitochondrial dehydrogenase. HL-60 cells (1 x 103 cells/well) were incubated at 37℃ in 96-well plates in 10% FCS medium and incubated with vehi이e or drugs for 24 or 48 hours.
성능/효과
In conclusion, the PPARy ligand, ciglitazone, and the RXR ligand, RA, can synergistically upregulate PTEN expression in human APL cells, resulting in inhibition of cell growth and cell cycle progression of acute leukemic cells associated with its ability to induce cell cycle arrest. Thus, the activation of the PPARy: RXR heterodimer might represent a regulatory pathway for HL- 60 cells and there may be a possible role for PPARy and RXR ligand in prophylactic and therapeutic approaches for controlling leukemia through the upregulation of PTEN, particularly to overcome RA-resistant APL, but also for other more common leukemias and types of cancers.
Moreover, upregulation of PTEN was significantly enhanced by a combination of both ciglitazone and RA. This is the first report to show that PPARy ligand, ciglitazone, and the RXR ligand, RA, synergistically enhance the upregulation of the tumor suppressor PTEN and inhibition of cell growth in human promyeloid leukemia cells, even though it need to be confirmed in the patients with APL.
참고문헌 (49)
Myers, M.P., Pass, I., and Batty, I.H. The lipid phosphatase activity of PTEN is critical for its tumor suppressor function. Proc. Natl. Acad. Sci. U. S. A. 95, 13513-13518 (1998)
Wen, S., Stolarov, J., and Myers, M.P. PTEN controls tumor-induced angiogenesis. Proc Natl. Acad.Sci. U. S. A. 98, 4622-4627 (2001)
Kim, J.S., Peng, X., De, P.K. PTEN controls immunoreceptor (immunoreceptor tyrosine-based activation motif) signaling and the activation of Rac. Blood 99, 694-697 (2002)
Li, J., Yen, C., and Liaw, D. PTEN, a putative protein tyrosine phosphatase gene mutated in human brain, breast, and prostate cancer. Science 275, 1943-1947 (1997)
Obata, K., Morland, S.J., and Watson, R.H. Frequent PTEN/MMAC mutations in endometrioid but not serous or mucinous epithelial ovarian tumors. Cancer Res. 58, 2095-2097 (1998)
Vlietstra, R.J., van Alewijk, D.C., Hermans, K.G., van Steenbrugge, G.J., and Trapman, J. Frequent inactivation of PTEN in prostate cancer cell lines and xenografts. Cancer Res. 58, 2720-2723 (1998)
Dahia, PL, Aguiar, RC, and Alberta J, et al. PTEN is inversely correlated with the cell survival factor Akt/PKB and is inactivated via multiple mechanismsin haematological malignancies. Hum. Mol. Genet. 8, 185-193 (1999)
Stambolic, V., Suzuki, A., and de la Pompa, J.L. Negative regulation of PKB/Akt-dependent cell survival by the tumor suppressor PTEN. Cell 95, 29-39 (1998)
Breitman, T.R., Selonick, S.E., and Collins, S.J. Induction of differentiation of the human promyelocytic leukemia cell line (HL-60) by retinoic acid. Proc. Natl. Acad. Sci. U. S. A. 77, 2936-2940 (1980)
Olsson, I.L., and Breitman, T.R. Induction of differentiation of the human histiocytic lymphoma cell line U-937 by retinoic acid and cyclic adenosine 3':5'-monophosphate-inducing agents. Cancer Res. 42, 3924-3927 (1982)
Barbui, T., Finazzi, G., and Falanga, A. The impact of alltrans-retinoic acid on the coagulopathy of acute promyelocytic leukemia. Blood 91, 3093-3102 (1998)
Li, Y., Dawson, M.I., and Agadir, A. Regulation of RAR beta expression by RAR- and RXR-selective retinoids in human lung cancer cell lines: effect on growth inhibition and apoptosis induction. Int. J. Cancer. 75, 88-95 (1998)
Rosen, E.D., and Spiegelman, B.M. PPARgamma: a nuclear regulator of metabolism, differentiation, and cell growth. J. Biol. Chem. 276, 37731-37734 (2001)
Konopleva, M., and Andreeff, M.. Role of peroxisome proliferator-activated receptor-gamma in hematologic malignancies. Curr. Opin. Hematol. 9, 294-302 (2002)
Kliewer, S.A., Umesono, K., Noonan, D.J., Heyman R.A., and Evans, R.M. Convergence of 9-cis retinoic acid and peroxisome proliferator signalling pathways through heterodimer formation of their receptors. Nature 358, 771-774 (1992)
Lehmann, J.M., Lenhard, J.M, and Oliver, B.B. Peroxisome proliferator-activated receptors alpha and gamma are activated by indomethacin and other non-steroidal anti-inflammatory drugs. J. Biol. Chem. 272, 3406-3410 (1997)
Warrell, RP, Jr., Frankel, SR, and Miller, W.H. Differentiation therapy of acute promyelocytic leukemia with tretinoin (all-trans-retinoic acid). N Engl J Med 324, 1385-1393 (1991)
Tontonoz, P., Singer S., and Forman, B.M. Terminal differentiation of human liposarcoma cells induced by ligands for peroxisome proliferator-activated receptor gamma and the retinoid X receptor. Proc. Natl. Acad. Sci. U. S. A. 94, 237-241 (1997)
Demetri, G.D., Fletcher, C.D., and Mueller, E. Induction of solid tumor differentiation by the peroxisome proliferator-activated receptor-gamma ligand troglitazone in patients with liposarcoma. Proc. Natl. Acad. Sci. U. S. A. 96, 3951-3956 (1999)
Tontonoz, P., Nagy, L., and Alvarez, J.G. PPARgamma promotes monocyte/macrophage differentiation and uptake of oxidized LDL. Cell 93, 241-252 (1998)
Asou, H., Verbeek, W., and Williamson, E. Growth inhibition of myeloid leukemia cells by troglitazone, a ligand for peroxisome proliferator activated receptor gamma, and retinoids. Int. J. Oncol. 15, 1027-1031 (1999)
Cornic, M., Delva, L, and Guidez, F. Induction of retinoic acid-binding protein in normal and malignant human myeloid cells by retinoic acid in acute promyelocytic leukemia patients. Cancer Res. 52, 3329-3334 (1992)
Brown, G, Bunce, CM, Rowlands, DC, et al. All-trans retinoic acid and 1 alpha, 25-dihydroxyvitamin D3 co-operate to promote differentiation of the human promyeloid leukemia cell line HL60 to monocytes. Leukemia 8, 806-815 (1994)
Witcher, M., Shiu, H.Y., Guo, Q., Miller, WH, Jr. Combination of retinoic acid and tumor necrosis factor overcomes the maturation block in a variety of retinoic acidresistant acute promyelocytic leukemia cells. Blood 104, 3335-3342 (2004)
Patel, L, Pass, I, Coxon, P. Tumor suppressor and antiinflammatory actions of PPARgamma agonists are mediated via upregulation of PTEN. Curr. Biol. 11, 764-768 (2001)
Nolte, RT, Wisely, GB, Westin, S. Ligand binding and coactivator assembly of the peroxisome proliferator-activated receptor-gamma. Nature 395, 137-143 (1998)
Kwak, Y.G., Song, C.H., Yi, H.K. Involvement of PTEN in airway hyperresponsiveness and inflammation in bronchial asthma. J. Clin. Invest. 111, 1083-1092 (2003)
Hwang, P.H., Yi, H.K, Kim, D.S., Nam, S.Y., Kim, J.S., and Lee, DY. Suppression of tumorigenicity and metastasis in B16F10 cells by PTEN/MMAC1/TEP1 gene. Cancer Lett. 172, 83-91 (2001)
Wightman, J., Roberson, M.S., Lamkin, T.J. Retinoic acid-induced growth arrest and differentiation: retinoic acid up-regulates CD32 (Fc gammaRII) expression, the ectopic expression of which retards the cell cycle. Mol. Cancer Ther. 1, 493-506, (2002)
Yang, Z., Bagheri-Yarmand, R., and Balasenthil, S. HER2 regulation of peroxisome proliferator-activated receptor gamma (PPARgamma) expression and sensitivity of breast cancer cells to PPARgamma ligand therapy. Clin. Cancer Res. 9, 3198-3203 (2003)
Suh, N., Wang, Y., and Honda, T. A novel synthetic oleanane triterpenoid, 2-cyano-3,12-dioxoolean-1,9-dien-28-oic acid, with potent differentiating, antiproliferative, and anti-inflammatory activity. Cancer Res. 59, 336-341 (1999)
Konopleva, M., Tsao, T., and Ruvolo, P. Novel triter penoid CDDO-Me is a potent inducer of apoptosis and differentiation in acute myelogenous leukemia. Blood 99, 326-335 (2002)
Denzlinger, C. D.K, and Mohle, A. Peroxisom proliperator activated receptor g (PPAR ${\gamma}$ ) agonist as antileukemic agents in CMML. Blood 98, 625a (2001)
Cantley, L.C., and Neel, B.G.. New insights into tumor suppression: PTEN suppresses tumor formation by restraining the phosphoinositide 3-kinase/AKT pathway. Proc. Natl. Acad .Sci. U. S. A. 96, 4240-4245 (1999)
Di Cristofano, A., and Pandolfi, P.P. The multiple roles of PTEN in tumor suppression. Cell 100, 387-390 (2000)
Cappellini, A., Tabellini, G., and Zweyer, M. The phosphoinositide 3-kinase/Akt pathway regulates cell cycle progression of HL60 human leukemia cells through cytoplasmic relocalization of the cyclin-dependent kinase inhibitor p27(Kip1) and control of cyclin D1 expression. Leukemia 17, 2157-2167 (2003)
Seminario, M.C., Precht, P. Wersto, R.P., Gorospe, M., and Wange, R.L. PTEN expression in PTEN-null leukaemic T cell lines leads to reduced proliferation via slowed cell cycle progression. Oncogene 22, 8195-8204 (2003)
Lee, YR, Shim, HJ, and Yu, H.N. Dimethylsulfoxide induces upregulation of tumor suppressor protein PTEN through nuclear factor-kappa B activation in HL-60 cells. Leuk. Res. 29, 401-405 (2005)
※ AI-Helper는 부적절한 답변을 할 수 있습니다.