The Role of Cdk5 in Retinoic Acid-Induced Apoptosis of Cervical Cancer Cell Line

10.4077/CJP.2009.AMG067

Hung-Shou Kuo；Fu-Ning Hsu；Ming-Ching Chiang；Shuen-Chi You；Mei-Chih Chen；Ming-Jae Lo；Ho Lin

retinoic acid ； apoptosis ； Cdk5 ； p35 ； HeLa cells

The Chinese Journal of Physiology

52卷1期（2009 / 02 / 01）

23 - 30

英文

Cdk5 is a small serine/threonine protein kinase which belongs to Cdk family. Unlike other Cdk members, so far Cdk5 is known to be irrelevant in cell cycle. Cdk5 kinase activity is regulated by binding with its activator, p35. Our previous results indicate that Cdk5 and p35 are involved in drugs-induced apoptosis of prostate cancer cells. Retinoic acid (RA) is one of the vitamin A-related compounds. Because of its potency on biological functions, it has been widely studied in its novel actions including the ability to inhibit cancer cell growth and to induce apoptosis. Here, we report that RA treatment decreased the growth of human cervical cancer cell line, HeLa, and Cdk5 contributed to this effect. The involvement of Cdk5 in RA-reduced cell survival was performed by treatments of Cdk5 inhibitor and siRNA. We further identified that RA-induced growth inhibition was partly correlated to Cdk5 activityrelated apoptosis by detecting cell cycle distribution of sub G1 phase and the signals of Annexin V staining. In addition, our results also indicated that Cdk5 activity was involved in RA-induced HeLa apoptosis by detecting cleavages of caspase-3 and its substrate, PARP (poly (ADP-ribose) polymerases) Interestingly, the nuclear localizations of Cdk5 and p35 proteins were increased by RA treatment, which, again, suggests the involvement of Cdk5 and p35 in RA-induced apoptotic effects. In conclusion, we provide evidence to suggest that Cdk5 and p35 might play important roles in RA-induced HeLa apoptosis.

1. Ahuja, H.S.,Szanto, A.,Nagy, L.,Davies, P.J.(2003).The retinoid X receptor and its ligands: versatile regulators of metabolic function, cell differentiation and cell death.J. Biol. Regul. Homeost. Agents,17,29-45.
2. Alvarez, A.,Munoz, J.P.,Maccioni, R.B.(2001).A Cdk5-p35 stable complex is involved in the β-amyloid-induced deregulation of Cdk5 activity in hippocampal neurons.Exp. Cell Res.,264,266-274.
3. Brown, A.M.,Riddoch, F.C.,Robson, A.,Redfern, C.P.,Cheek, T.R.(2005).Mechanistic and functional changes in Ca2+ entry after retinoic acid-induced differentiation of neuroblastoma cells.Biochem. J.,388,941-948.
4. Chergui, K.,Svenningsson, P.,Greengard, P.(2004).Cyclin-dependent kinase 5 regulates dopaminergic and glutamatergic transmission in the striatum.Proc. Natl. Acad. Sci. U.S.A.,101,2191-2196.
5. Chomienne, C.,Fenaux, P.,Degos, L.(1996).Retinoid differentiation therapy in promyelocytic leukemia.FASEB J.,10,1025-1030.
6. Geisen, C.,Denk, C.,Kupper, J.H.,Schwarz, E.(2000).Growth inhibition of cervical cancer cells by the human retinoic acid receptor β gene.Int. J. Cancer,85,289-295.
7. Hong, W.K.,Lippman, S.M.,Itri, L.M.,Karp, D.D.,Lee, J.S.,Byers, R.M.,Schantz, S.P.,Kramer, A.M.,Lotan, R.,Peters, L.J.,Dimery, I.W.,Brown, B.W.,Goepfert, H.(1990).Prevention of second primary tumors with isotretinoin in squamous-cell carcinoma of the head and neck.N. Engl. J. Med.,323,795-801.
8. Hsu, S.L.,Chen, M.C.,Chou, Y.H.,Hwang, G.Y.,Yin, S.C.(1999).Induction of p21(CIP1/Waf1) and activation of p34 (cdc2) involved in retinoic acid-induced apoptosis in human hepatoma Hep3B cells.Exp. Cell Res.,248,87-96.
9. Kerokoski, P.,Suuronen, T.,Salminen, A.,Soininen, H.,Pirttila, T.(2001).The levels of cdk5 and p35 proteins and tau phosphorylation are reduced during neuronal apoptosis.Biochem. Biophys. Res. Commun.,280,998-1002.
10. Kraemer, K.H.,DiGiovanna, J.J.,Moshell, A.N.,Tarone, R. E.,Peck, G. L.(1988).Prevention of skin cancer in xeroderma pigmentosum with the use of oral isotretinoin.N. Engl. J. Med.,318,1633-1637.
11. Kwon, Y.T.,Gupta, A.,Zhou, Y.,Nikolic, M.,Tsai, L.H.(2000).Regulation of N-cadherin-mediated adhesion by the p35-Cdk5 kinase.Curr. Biol.,10,363-372.
12. Lee, S.Y.,Wenk, M.R.,Kim, Y.,Nairn, A.C.,De Camilli, P.(2004).Regulation of synaptojanin 1 by cyclin-dependent kinase 5 at synapses.Proc. Natl. Acad. Sci. U.S.A.,101,546-551.
13. Lew, J.,Beaudette, K.,Litwin, C.M.,Wang, J.H.(1992).Purification and characterization of a novel proline-directed protein kinase from bovine brain.J. Biol. Chem.,267,13383-13390.
14. Lin, H.,Chen, M.C.,Chiu, C.Y.,Song, Y.M.,Lin, S.Y.(2007).Cdk5 regulates STAT3 activation and cell proliferation in medullary thyroid carcinoma cells.J. Biol. Chem.,282,2776-2784.
15. Lin, H.,Juang, J.L.,Wang, P.S.(2004).Involvement of Cdk5/p25 in digoxin-triggered prostate cancer cell apoptosis.J. Biol. Chem.,279,29302-29307.
16. Lin, H.,Lin, T.Y.,Juang, J.L.(2007).Abl deregulates Cdk5 kinase activity and subcellular localization in Drosophila neurodegeneration.Cell. Death Differ.,14,607-615.
17. Lotan, R.(1995).Retinoids and apoptosis: implications for cancer chemoprevention and therapy.J. Natl. Cancer Inst.,87,1655-1657.
18. Mark, M.,Ghyselinck, N.B.,Chambon, P.(2006).Function of retinoid nuclear receptors: lessons from genetic and pharmacological dissections of the retinoic acid signaling pathway during mouse embryogenesis.Annu. Rev. Pharmacol. Toxicol.,46,451-480.
19. McCaffery, P.,Zhang, J.,Crandall, J.E.(2006).Retinoic acid signaling and function in the adult hippocampus.J. Neurobiol.,66,780-791.
20. Neystat, M.,Rzhetskaya, M.,Oo, T.F.,Kholodilov, N.,Yarygina, O.,Wilson, A.,El-Khodor, B.F.,Burke, R.E.(2001).Expression of cyclin-dependent kinase 5 and its activator p35 in models of induced apoptotic death in neurons of the substantia nigra in vivo.J. Neurochem.,77,1611-1625.
21. Nikolic, M.,Dudek, H.,Kwon, Y.T.,Ramos, Y.F.,Tsai, L.H.(1996).The cdk5/p35 kinase is essential for neurite outgrowth during neuronal differentiation.Genes Dev.,10,816-825.
22. Paglini, G.,Pigino, G.,Kunda, P.,Morfini, G.,Maccioni, R.,Quiroga, S.,Ferreira, A.,Caceres, A.(1998).Evidence for the participation of the neuron-specific CDK5 activator P35 during laminin-enhanced axonal growth.J. Neurosci.,18,9858-9869.
23. Piacentini, M.,Annicchiarico-Petruzzelli, M.,Oliverio, S.,Piredda, L.,Biedler, J.L.,Melino, E.(1992).Phenotype-specific "tissue" transglutaminase regulation in human neuroblastoma cells in response to retinoic acid: correlation with cell death by apoptosis.Int. J. Cancer,52,271-278.
24. Smith, M.A.,Parkinson, D.R.,Cheson, B.D.,Friedman, M.A.(1992).Retinoids in cancer therapy.J. Clin. Oncol.,10,839-864.
25. Um, S.J.,Kim, E.J.,Hwang, E.S.,Kim, S.J.,Namkoong, S.E.,Park, J.S.(2000).Antiproliferative effects of retinoic acid/interferon in cervical carcinoma cell lines: cooperative growth suppression of IRF-1 and p53.Int. J. Cancer,85,416-423.
26. Zhang, J.,Krishnamurthy, P.K.,Johnson, G.V.(2002).Cdk5 phosphorylates p53 and regulates its activity.J. Neurochem.,81,307-313.

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