Tumor necrosis ftctor-alpha$(TNF-{\alpha})$는 세포의 고사, 염증 및 면역 등의 다양한 생물학적 기능에 대한 역할을 한다. PTEN 역시 세포의 성장과 증식 그리고 세포의 유주와 분화 등의 세포학적인 다양한 기능을 갖는다 그러므로 이들 두 분자들 사이의 상호관계가 있을 것으로 제안되고 있으며, $TNF-{\alpha}$는 사람의 대장세포 주인 HT-29에서 nuclear factor-kappa $B(NF-{\kappa}B)$ 경로를 통해 PTEN downregulate 기능이 있는 것으로 알려져 왔다. 그러나 저자 등은 본 연구에서 HL-60 cells에서 $TNF-{\alpha}$가 $NF-{\kappa}B$를 통해 PTEN를 upregulates하는 기존의 반대 현상을 확인하였다. $TNF-{\alpha}$는 HL-60 cells에서 time과 dose의존성 방법으로 PTEN 발현을 증가시켰지만 반응은 p65 anisense oligonucleotide 또는 pyrrolidine dithiocarbamate(PDTC)으로 $NF-{\kappa}B$를 분해함으로 파괴되었다. 따라서 저자 등은 $TNF-{\alpha}$가 $NF-{\kappa}B$경로를 활성화시킴을 확인하였고, $TNF-{\alpha}$를 처리 할 경우 핵에 대하여 p65 전위에 의해 $TNF-{\alpha}$가 활성화됨을 증명하였다. 결국 HL-60세포에서 $NF-{\kappa}B$의 활성화에 따라 PTEN 발현의 upregulation이 유도되는 것으로 결론지었다.
Tumor necrosis ftctor-alpha$(TNF-{\alpha})$는 세포의 고사, 염증 및 면역 등의 다양한 생물학적 기능에 대한 역할을 한다. PTEN 역시 세포의 성장과 증식 그리고 세포의 유주와 분화 등의 세포학적인 다양한 기능을 갖는다 그러므로 이들 두 분자들 사이의 상호관계가 있을 것으로 제안되고 있으며, $TNF-{\alpha}$는 사람의 대장세포 주인 HT-29에서 nuclear factor-kappa $B(NF-{\kappa}B)$ 경로를 통해 PTEN downregulate 기능이 있는 것으로 알려져 왔다. 그러나 저자 등은 본 연구에서 HL-60 cells에서 $TNF-{\alpha}$가 $NF-{\kappa}B$를 통해 PTEN를 upregulates하는 기존의 반대 현상을 확인하였다. $TNF-{\alpha}$는 HL-60 cells에서 time과 dose의존성 방법으로 PTEN 발현을 증가시켰지만 반응은 p65 anisense oligonucleotide 또는 pyrrolidine dithiocarbamate(PDTC)으로 $NF-{\kappa}B$를 분해함으로 파괴되었다. 따라서 저자 등은 $TNF-{\alpha}$가 $NF-{\kappa}B$경로를 활성화시킴을 확인하였고, $TNF-{\alpha}$를 처리 할 경우 핵에 대하여 p65 전위에 의해 $TNF-{\alpha}$가 활성화됨을 증명하였다. 결국 HL-60세포에서 $NF-{\kappa}B$의 활성화에 따라 PTEN 발현의 upregulation이 유도되는 것으로 결론지었다.
Tumor necrosis factor-alpha $(TNF-{\alpha})$ plays a variety of biological functions such as apoptosis, inflammation and immunity. PTEN also has various cellular function including cell growth, proliferation, migration and differentiation. Thus, possible relationships between two molecule...
Tumor necrosis factor-alpha $(TNF-{\alpha})$ plays a variety of biological functions such as apoptosis, inflammation and immunity. PTEN also has various cellular function including cell growth, proliferation, migration and differentiation. Thus, possible relationships between two molecules are suggested. $(TNF-{\alpha})$has been known to downregulate PTEN via nuclear factor-kappa $B(NF-{\kappa}B)$ pathway in the human colon cell line, HT-29. However, here we show the opposite finding that $(TNF-{\alpha})$ upregulates PTEN via activation of $NF-{\kappa}B$ in HL-60 cells. $TNF-{\alpha}$ increased PTEN expression at HL-60 cells in a time- and dose-dependent manner, but the response was abolished by disruption of $NF-{\kappa}B$ with p65 anisense oligonucleotide or pyrrolidine dithiocarbamate (PDTC). We found that $TNF-{\alpha}$ activated the $NF-{\kappa}B$ pathways, evidenced by the translocation of p65 to the nucleus in $TNF-{\alpha}-treated$ cells. We conclude that $TNF-{\alpha}$ induces upregulation of PTEN expression through $NF-{\kappa}B$ activation in HL-60 cells.
Tumor necrosis factor-alpha $(TNF-{\alpha})$ plays a variety of biological functions such as apoptosis, inflammation and immunity. PTEN also has various cellular function including cell growth, proliferation, migration and differentiation. Thus, possible relationships between two molecules are suggested. $(TNF-{\alpha})$has been known to downregulate PTEN via nuclear factor-kappa $B(NF-{\kappa}B)$ pathway in the human colon cell line, HT-29. However, here we show the opposite finding that $(TNF-{\alpha})$ upregulates PTEN via activation of $NF-{\kappa}B$ in HL-60 cells. $TNF-{\alpha}$ increased PTEN expression at HL-60 cells in a time- and dose-dependent manner, but the response was abolished by disruption of $NF-{\kappa}B$ with p65 anisense oligonucleotide or pyrrolidine dithiocarbamate (PDTC). We found that $TNF-{\alpha}$ activated the $NF-{\kappa}B$ pathways, evidenced by the translocation of p65 to the nucleus in $TNF-{\alpha}-treated$ cells. We conclude that $TNF-{\alpha}$ induces upregulation of PTEN expression through $NF-{\kappa}B$ activation in HL-60 cells.
* AI 자동 식별 결과로 적합하지 않은 문장이 있을 수 있으니, 이용에 유의하시기 바랍니다.
문제 정의
O니r result is 什le first evidence that PTEN is target of TNF- cc/NF-kB pathways in le니kemia cells. This paper might help us to understand the mechanism by which TNF-a induces differentiation of leukemia cells.
가설 설정
These results suggest that TNF, strong NF-kB activator, could affect the PI3K/Akt signaling pathway in HL-60 cells. Therefore, it was hypothesized that “the expression of PTEN may be upregulated by TNF, ?.
TNF-a activates I-kB kinase (IKK) that phosphorylates I-kB% causing I-KBa degradation by proteasome and allowing NF-kB to translocate to the nucleus to activate transcription 37-38). To test whether TNF-a activates NF-kB in HL-60 cells, we measured the translocation of NF・kB p65 subunit from cytosol to nucleus, and the level of I-KBa in cytosol. The translocation of p65 was clearly enhanced from 3 hr after the treatment of TNF-a and the enhancement persisted till 6hr (Fig.
제안 방법
4 μM of each PCR primer: sense primer, human PTEN (5'-CCG- GAATTCATGACAGCCArCArCAAAGA-3, )> antisense primer, human PTEN (5, -CGCGGATCCTCAGACTTT- TGTAATTTGTG-3'). Amplification for PTEN was initiated with 3 minutes of denaturation at 94℃ followed by 30 cycles at 94℃ for 1 minute, 94℃ for 1 minute, 55℃ for 1 minute, and 72℃ for 2 minutes. After the last cycle of amplification, the samples were incubated for 5 minutes at 72℃.
대상 데이터
Louis, MO, USA). 5' end p65 antisense oligonucleotide of NF-kB gene (5'・GAAACAGATCGTCCATGGT・3‘)and p65 nonsense (scrembled control) oligonucleotide (5f-GTAC- TACTCTGAGCAAGGA-3') was designed and manufactured by GenoTech (Daejon, Korea).
The anti-PTEN monoclonal mouse antibody and The anti p65 monoclonal mouse antibody were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and the phospho-AKT (p-AKT) polyclonal rabbit antibody from Cell Signaling Technology (Minneapolis, MN, USA). Recombinant human cytokine, tumor necrosis factor-a (TNF-a) was obtained from R & D Company (Minneapolis, MN, USA).
참고문헌 (37)
Vivanco, I. and Sawyers, C.L. The phosphatidylinositol 3-Kinase AKT pathway in human cancer. Nat. Rev. Cancer 2, 489-501 (2002)
Maehama, T., and Dixon, J.E. The tumor suppressor, PTEN/MMAC1, dephosphorylates the lipid second messenger, phosphatidylinositol 3,4,5,-triphosphate. J. Biol. Chem. 273, 13375-13378 (1998)
Myers, M.P., Pass, I., Batty, I.H., Van der Kaay, J., Stolarov, J.P., Hemmings, B.A., Wigler, M.H., Downes, C.P., and Tonks, N.K. The lipid phosphatase activity of PTEN is critical for its tumor suppressor function. Proc. Natl. Acad. Sci. U.S.A. 95, 13513-13518 (1998)
Tonks, N.K. and Myers, M.P. Structural assets of a tumor suppressor. Science 286, 2096-2097 (1999)
Samuels, Y. and Ericson, K. Oncogenic PI3K and its role in cancer. Curr. Opin. Oncol. 1, 77-82 (2006)
Hennessy, B.T., Smith, D.L., Ram, P.T., Lu, Y., and Mills, G.B. Exploiting the PI3K/AKT pathway for cancer drug discovery. Nat. Rev. Drug Discov. 4, 988-1004 (2005)
Lachyankar, M.B., Sultana, N., Schonhoff, C.M., Mitra, P., Poluha, W., Lambert, S., Quesenberry, P.J., Litofsky, N.S., Recht, L.D., Nabi, R., Miller, S.J., Ohta, S., Neel, B.G., and Ross, A.H. A role for nuclear PTEN in neuronal differentiation. J. Neurosci. 20, 1404-1413 (2004)
Kurose, K., Zhou, X.P., Araki, T., Cannistra, S.A., Maher, E.R., and Eng, C. Frequent loss of PTEN expression is linked to elevated phosphorylated Akt levels, but not associated with p27 and cyclin D1 expression, in primary epithelial ovarian carcinomas. Am. J. Pathol. 158, 2097-106 (2001)
Kim, S., Domon-Dell, C., Kang, J., Chung Dai H., Freund J., and Evers B.M. Down-regulation of the tumor suppressor PTEN by the tumor necrosis factor-/nuclear factor- ${\kappa}B$ (NF- ${\kappa}B$ )-inducing kinase/NF- ${\kappa}B$ pathway is linked to a default I-eB-autoregulatory loop. J. Biol. Chem. 279, 4285-4291 (2004)
Lee, Y.R, Shim, H.J., Yu, H.N., Song, E.K., Park, J., Kwon, K.B., Park, J.W., Rho, H.W., Park, B.H., Han, M.K., and Kim, J.S. Dimethylsulfoxide induces upregulation of tumor suppressor protein PTEN through nuclear factor-kappaB activation in HL-60 cells. Leuk. Res. 29, 401-405 (2005)
Chae, G.N. and Kwak, S.J. NF- ${\kappa}B$ is involved in the TNF-a induced inhibition of the differentiation of 3T3-L1 cells by reducing PPARg expression. Exp. Mol. Med. 35, 431-437 (2003)
Hsu, H., Huang, J., Shu, H.B., Baichwal, V., and Goeddel, D.V. TNF-dependent recruitment of the protein kinase RIP to the TNF receptor-1 signaling complex. Immunity 4, 387-396 (1996)
Chen, C., Edlstein, L.C., and Gelinas, C. The Rel/NF-kB family directly activates expression of the apoptosis inhibitor Bcl-x(L). Mol. Cell. Biol. 20, 2687-2695 (2000)
Wang, C.Y., Mayo, M.W., Korneluk, R.G., Goeddel, D.V., and Baldwin, Jr A.S. NF-kappaB antiapoptosis: induction of TRAF1 and TRAF2 and c-IAP and c-IAP2 to suppress caspase-8 activation. Science 281, 1680-1683 (1998)
Wang, C.Y., Mayo, M.W., and Baldwin, Jr A.S. TNF-and cancer therapy-induced apoptosis: potenitation by inhibition of NF-kappaB. Science 274, 784-787 (1996)
Bradford, M.M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248-254 (1976)
Cappellini, A., Tabellini, G., Zweyer, M., Bortul, R., Tazzari, P.L., Billi, A.M., Fala, F., Cocco, L., and Martelli, A.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)
Altucci, L., Rossin, A., Raffelsberger, W., Reitmair, A., Chomienne, C., and Gronemeyer, H. Retinoic acid-induced apoptosis in leukemia cells is mediated by paracrine action of tumor-selective death ligand TRAIL. Nat. Med.7, 680-686 (2001)
DiDonato, J.A., Hayakawa, M., Rothwarf, D.M., Zandi, E., and Karin, M. A cytokine-responsive IkappaB kinase that activates the transcription factor NF-kappaB. Nature 388, 548-554 (1997)
Mercurio, F., Zhu, H., Murray, B.W., Shevchenko, A., Bennett, B.L., Li, J., Young, D.B., Barbosa, M., Mann, M., Manning, A., and Rao, A. IKK-1 and IKK-2: cytokineactivated IkappaB kinases essential for NF-kappaB activation. Science 278, 860-866 (1997)
※ AI-Helper는 부적절한 답변을 할 수 있습니다.