Toxicological Research

Korean Society of Toxicology & Korea Environmental Mutagen Society (한국독성학회)
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Apoptotic Signaling Pathway by Cadmium in Hepalclc7 cells

Hepa1c1c7 세포에서 카드뮴에 의한 세포사멸 신호전달체계에 관한 연구

  • 오경재 (전북대학교 의과대학 예방의학교실) ;
  • 염정호 (전북대학교 의과대학 예방의학교실)
  • Published : 2001.10.01
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Abstract

Cadmium is an ubiquitous toxic metal and chronic exposure to cadmium results in the accumulation of cadmium in the liver and kidneys. In contrast, acute exposure leads to damage mainly in the liver. Apoptosis induced by cadmium has been shown in many tissues in vivo and in cultured cells in vitro. However, the molecular mechanism of cadmium-induced apoptosis is not clear in hepatocyte. To investigate the induction of apoptosis in the hepatocyte, we used mouse hepatoma cell line, Hepalclc7 cells, and analysed the molecules that involved in cadmium-induced apoptosis. Cadmium induced the genomic DNA fragmentation, PARP cleavage, and activation of caspase-3 like protease. Caspase-9 cysteine protease was activated in a time-dependent manner but caspase-8 cysteine protease was not significantly activated in cadmium-treated Hepalclc7 cells. Cadmium also induced mitochondrial dysfunction including cytochrome c release from mitochondria, change oj mitochondrial membrane potential tranition, and tranlocation of Bax Protein into mitochondria. These results strong1y indicated that the signal Pathway of apoptotic death in cadmium-treated Hepalclc7 cells is modulated by caspase cascade via mitochondria.

카드뮴의 주요한 표적장기이며 카드뮴이 만성 및 급성 폭로시 축적되는 가장 중요한 장기인 간의 세포독성을 Hepalclc7세포에서 caspases및 Bax단백질의 활성과 발현 그리고 미토콘드리아 세포막 전위 변화(MPT) 등을 조사하여 다음과 같은 결과를 얻었다. 1. 카드뮴은 농도의존적으로 간세포인 Hepalclc7 세포의 생존율을 감소시켰다. 2. 카드뮴을 농도별로 처리하였을 때 100 M 이상의 농도에서 세포사멸의 특징중의 하나인 DNA분절현상을 확인하였다. 3. 카드뮴 처리 후 caspase-3, caspase-8, caspase-9 의 활성변화를 조사한 결과 caspase-3,-9 pretease 활성이 시간이 경과함에 따라 증가하였다. 4. 카드뮴 처리 후 cytochrome c가 세포질내로 방출되었고 이는 caspase-9 proteas의 활성화를 유도하였다. 5. 카드뮴 처리 후 Bax가 세포질에서 미토콘드리아로 이동하여 cytochrome c의 세포질내로의 방출에 관여하였다. 6. 카드뮴 처리시 미토콘드리아 세포막 전위차의 감소를 JC-1 형광염색을 통하여 확인하였다. 이상의 결과는 카드뮴에 의한 Hepalclc7 세포사멸의 신호전달기전은 세포질내에 있는 Bax가 미토콘드리아로 이동, cytochrome c의 세포질내로의 방출, 그리고 caspase-3, 9 pretease 활성화를 의해서 매개되는 것으로 판단된다. 또한 Bax 단백질의 발현변화가 미토콘드리아의 기능변화에 기여하였으리라 사료된다.

Keywords

References

  1. Cancer Res. v.58 Identification and characterrization of murine capase-14. a new member of the caspase family Ahmad, M.;Srinivasula, S. M.;Hedge, R.;Mukattash, R.;Rernandes-Alnemri, T.;Alnemri, E. S.
  2. CMLS v.54 Mechanism controlling cellular suicide: role of Bcl-2 and caspases Allen, R. T.;Cluck, M. W.;Agrawal, D. K.
  3. Cell v.87 Human ICE CED-3 protease nomenclaure Alnemri, E. S.;Livingston, D. J.;Nicholson, D. W.;Savesen, G.;Thornberry, N. A.;Wong, W. W.;Yuan, J.
  4. Cell v.41 Cloning the chromosomal breakpoint of t(14;18) human lymphona: clustering around JH on chromosone 14 and near a transcroptional unit on 18 Bakhshi, A.;Jensen, J. P.;Goldman, P.;Wright, J. J.;McBride, O. W.;Epstein, A. L.;Korsmeyer, S. J.
  5. Proc. Natl. Acad. Sci. v.82 Nucleotide sequence of a t(14;18) chromosomal breakpoint in follicular lymphoma and demonstration of a breakpoint -cluster region near a transcriptionally active locus on chromosome 18 Cleary, M. L.;Sklar, J.
  6. Toxicol. Appl. Pharmacol. v.65 Acute exposure to cadmium causes severe liver injury in rats Dudley, R. E.;Svoboda, D. J.;Klassen, C. D.
  7. Toxicol. Appl. Pharmacol. v.76 Time-course of cadmium-induced ultrastructural changes in rat liver Dudley, R. E.;Svoboda, D. J.;Klaassen, D. D.
  8. Genes Dev. v.13 Bcl-2 family members and the mitochondria in apoptosis Gross, A.;McDonnell, J. M.;Korsmeyer, S. J.
  9. EMBO J. v.16 Action of CD95 (APO-1/Fas) signaling by ceramice mediates cancer therapy-induced apoptosis Herr, I.;Wilhelm, D.;Bohler, T.;Angel, P.;Debatin, K. M.
  10. Nature v.348 Bcl-2 is an inner mitochondrial membrane protein that blocks programmed cell death Hockenbery, D. M.;Nunez, G.;Milliman, C.;Schreiber, R. D.;Korsmeyer, S. J.
  11. Casarett and Doull's Toxicology Klassen, C. D.;Amdur, M. O.;Doull, J.
  12. Trends Genet v.11 Regulators of cell death Korsmeyer, S. J.
  13. Toxicol. Appl. Pharmacol. v.41 Toxicity and distribution of cadmium administered to rats at sublethal doses Kotsonis, F. N.;Klaassen, C. D.
  14. EMBO J. v.18 no.21 53 regulates mirochondrial membrane potential through reactive oxygen species and induces cytochrome c-independent apoptosis blocked by Bcl-2 Lipf, Dietz R.;Von Harsdorf. R.
  15. Biochem Pharmacol v.47 no.9 Selenite induction of DNA strand breaks and apoptosis in mouse leukemic L1210 cells Lu, J.;Kaeck, M.;Jiang, C.;Wilson, A. C.;Tompson, H. J.
  16. Am. J. Pathol. v.142 no.6 Role of apoprosis in the rat Rao, M. S.;Yelandandi, A. V.;Subbarao, V.;Reddy, J. K.
  17. J. Cell Biol. v.124 Bcl-2 and the regulation of programmed cell death Reed, J. C.
  18. Eur. J. Biochem. v.254 Apoptosis signaling by death receptors Schulze-Osthoff, K.;Ferrari, D.;Los, M.;Wesselborg, S.;Pater, M. E.
  19. Arch. Emviron. Environ. Health v.24 Biological differences in cadmium and zinc turnover Shaikh, Z. A.;Lucis, O. J.
  20. Toxicol. Appl. Pharmacol. v.143 Inductinon of apoptosis in human T-cells by organomercuric cpmpounds: a flow cytometric analysis Shenker, B. J.;Datar, S.;S. Mansfield, K.;Shapiro, I. M.
  21. Ann. Clin. Lab. Sci. v.25 no.2 The influence of zinc on apoptosis Sundrman, Jr.;F. W.
  22. Science v.228 Involvement of the bcl-2 gene in human follicular lymphoma Tsujimoto, Y.;Crossman, J.;Jaffe, E.;Croce, C. M.
  23. Cell v.28 Bcl-xL regulates the membrane potential and volume homeostasis of mitochondria Vander Heiden, M. G.;Chandel, N. S.;Williamson, E. K.;Schimacker, P. T.;Thompson, C. B.
  24. Nat. Cell Biol. v.1 Bcl-2 proteins: inhibitors of apoptosis or regulators of mitochondrial homeostasis? Vander Heiden, M. G.;Thompson, C. B.
  25. Nature v.335 Bcl-2 gene promotes haemopoietic cell survival and cooperates with c-myc to immortalize pre-B cells Vaux, D. L.;Cory, S.;Adams, J.
  26. Radioth Oncol. v.47 The role of the stress-activated kinase signaling pathway in radiation-induced apoptosis Verheij, M.;Ruiter, G. A.;Zerp, S. F.;van Bitterswijk, W. J.;Fuks, J.;Haimovitz-Freiderman, A.;Bartelink, H.
  27. JBC v.273 no.1 Induction of c-fos proto-oncogene in mesangial cells by cadmjum Wang, Z.;Templeton, D. M.
  28. J. Biol. Chem. v.273 no.12 Caspase-dependent cleavage of signaling proteins during apoptosis Widmann, C.;Gibson, S.
  29. Int. Rev. Cytol. v.68 Cell death: The significance of cell death Wyllie, A. H.;Kerr, J. F. R.;Currie, A. R.
  30. Science v.270 Opposing effect of ERK and JNK-p38 MAP kinases on apoptosis Xia, Z.;Dickens, M.;Raingaud, J.;Davis, R. J.;Greenberg, M. E.