第十號程序性細胞死亡分子 (programmed cell death 10, PDCD10, TFAR15) 屬於細胞凋亡相關基因家族成員之一。PDCD10首次被報導是當培養人類骨髓癌細胞株TF-1時，在缺乏粒細胞巨噬細胞集落刺激因子(granulocyte macrophage colony- stimulating factor, GM-CSF) 的狀態下，細胞會走向凋亡而造成PDCD10的基因表現量被提高。根據病理統計報導，PDCD10基因突變與顱內海綿狀血管瘤的發生率 (cerebral cavernous malformation, CCM) 極具正相關性，故此基因又被稱為CCM3 (cerebral cavernous malformation 3) 基因。根據研究報導指出，在培養人類急性骨髓白血病細胞株HL-60時，若缺乏胎牛血清的狀態下，PDCD10的基因表現量亦會提高，並可透過細胞外信號調節激酶 (extracellular signal-regulated kinase, ERK) 來促進細胞增生以及細胞轉化的現象。意外的是，根據本論文的研究，在缺乏胎牛血清的細胞培養狀態下，HL-60細胞的PDCD10基因表現量反倒呈現著下調的情形。此外，根據純化之第十號程序性細胞死亡分子重組蛋白，我們成功製備了PDCD10的多株抗體。利用此抗體，我們進一步証明了HL-60細胞在缺乏胎牛血清的培養狀態下，PDCD10的蛋白質表現量ㄧ樣是呈現著下調的情形。而另一方面當大量表現PDCD10蛋白質於神經膠質瘤細胞株U-87 MG時，可以發現到癌細胞的轉移能力受到明顯抑制，並且基質金屬蛋白酶9 (matrix metalloproteinase 9, MMP9) 的表現量與ERK的活性被上調增加。利用絲裂原活化蛋白激酶的激酶 (mitogen-activated protein kinase kinase, MAPKK) 抑制劑PD98059來抑制細胞內的ERK被活化時，可以觀察到MMP9的表現量減少。因此證明了PDCD10極有可能具有抑制細胞凋亡的功能，並藉由ERK來調控MMP9的表現量進而影響著細胞的侵襲轉移或凋亡。

Programmed cell death 10 (PDCD10, TFAR15) is a member of the PDCD family associated with cell apoptosis. It was originally identified in the human premyeloid TF-1 cells for being up-regulated, in which apoptosis was induced by withdrawal of granulocyte macrophage colony-stimulating factor (GM-CSF). Clinical statistics have shown that mutations of PDCD10 are correlated with cerebral cavernous malformation (CCM) and thus this gene is also named as CCM3 (cerebral cavernous malformation 3) accordingly. The expression of PDCD10 was reported to be up-regulated in human promyeloblast HL-60 cells after serum starvation. The elevated PDCD10 then modulates the ERK (extracellular signal-regulated kinase) signaling pathway to promote cell growth and cellular transformation. Surprisingly, in this thesis it is found that the PDCD10 expression is in fact down-regulated in serum-free culture condition. By using the purified PDCD10 recombinant protein as an antigen, I successfully generated mouse anti-human PDCD10 polyclonal antibody. With this new antibody, I was able to further clarify that the PDCD10 protein expression is down-regulated in HL-60 cells after serum starvation. Meanwhile, overexpression of PDCD10 not only significantly suppresses the invasive ability of human glioblastoma U-87 MG cells, but also increases the expression and secretion of MMP9 as well as the ERK activity. Treatment of mitogen-activated protein kinase kinase (MAPKK) inhibitor PD98059 resulted in a suppression of the ERK activity and subsequent decrease of the MMP9 expression. Together these results suggest that PDCD10 may be an anti-apoptotic gene which affects cell apoptotic process as well as tumor cell invasion and metastasis by regulating the expression of MMP9 via the ERK pathway.

Agata, Y., Kawasaki, A., Nishimura, H., Ishida, Y., Tsubata, T., Yagita, H., and Honjo, T. (1996). Expression of the PD-1 antigen on the surface of stimulated mouse T and B lymphocytes. Int Immunol 8, 765-772.
Aguirre, A. J., Brennan, C., Bailey, G., Sinha, R., Feng, B., Leo, C., Zhang, Y., Zhang, J., Gans, J. D., Bardeesy, N., et al. (2004). High-resolution characterization of the pancreatic adenocarcinoma genome. Proc Natl Acad Sci U S A 101, 9067-9072.
Annabi, B., Thibeault, S., Moumdjian, R., and Beliveau, R. (2004). Hyaluronan cell surface binding is induced by type I collagen and regulated by caveolae in glioma cells. J Biol Chem 279, 21888-21896.
Bergametti, F., Denier, C., Labauge, P., Arnoult, M., Boetto, S., Clanet, M., Coubes, P., Echenne, B., Ibrahim, R., Irthum, B., et al. (2005). Mutations within the programmed cell death 10 gene cause cerebral cavernous malformations. Am J Hum Genet 76, 42-51.
Chang, W. S., Chang, N. T., Lin, S. C., Wu, C. W., and Wu, F. Y. (2000). Tissue-specific cancer-related serpin gene cluster at human chromosome band 3q26. Genes Chromosomes Cancer 29, 240-255.
Chen, J., Tian, Y. Q., and Xiao, J. Y. (2005). [Difference expression of gene for laryngeal squamous cell carcinoma by cDNA microarray and signal pathways of cell cycle regulation and cell apoptosis]. Zhong Nan Da Xue Xue Bao Yi Xue Ban 30, 483-486.
Chen, P. Y., Chang, W. S., Chou, R. H., Lai, Y. K., Lin, S. C., Chi, C. Y., and Wu, C. W. (2007). Two non-homologous brain diseases-related genes, SERPINI1 and PDCD10, are tightly linked by an asymmetric bidirectional promoter in an evolutionarily conserved manner. BMC Mol Biol 8, 2.
Chou, R. H., Lin, K. C., Lin, S. C., Cheng, J. Y., Wu, C. W., and Chang, W. S. (2004). Cost-effective trapezoidal modified Boyden chamber with comparable accuracy to a commercial apparatus. BioTechniques 37, 724-726.
Coussens, L. M., Tinkle, C. L., Hanahan, D., and Werb, Z. (2000). MMP-9 supplied by bone marrow-derived cells contributes to skin carcinogenesis. Cell 103, 481-490.
Denier, C., Labauge, P., Bergametti, F., Marchelli, F., Riant, F., Arnoult, M., Maciazek, J., Vicaut, E., Brunereau, L., and Tournier-Lasserve, E. (2006). Genotype-phenotype correlations in cerebral cavernous malformations patients. Ann Neurol 60, 550-556.
Dubovsky, J., Zabramski, J. M., Kurth, J., Spetzler, R. F., Rich, S. S., Orr, H. T., and Weber, J. L. (1995). A gene responsible for cavernous malformations of the brain maps to chromosome 7q. Hum Mol Genet 4, 453-458.
Dupre, N., Verlaan, D. J., Hand, C. K., Laurent, S. B., Turecki, G., Davenport, W. J., Acciarri, N., Dichgans, J., Ohkuma, A., Siegel, A. M., and Rouleau, G. A. (2003). Linkage to the CCM2 locus and genetic heterogeneity in familial cerebral cavernous malformation. Can J Neurol Sci 30, 122-128.
Egeblad, M., and Werb, Z. (2002). New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer 2, 161-174.
Fujimura, M., Watanabe, M., Shimizu, H., and Tominaga, T. (2007). Expression of matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinase (TIMP) in cerebral cavernous malformations: immunohistochemical analysis of MMP-2, -9 and TIMP-2. Acta Neurochir (Wien) 149, 179-183; discussion 183.
Goffin, F., Munaut, C., Frankenne, F., Perrier D'Hauterive, S., Beliard, A., Fridman, V., Nervo, P., Colige, A., and Foidart, J. M. (2003). Expression pattern of metalloproteinases and tissue inhibitors of matrix-metalloproteinases in cycling human endometrium. Biol Reprod 69, 976-984.
Guclu, B., Ozturk, A. K., Pricola, K. L., Bilguvar, K., Shin, D., O'Roak, B. J., and Gunel, M. (2005). Mutations in apoptosis-related gene, PDCD10, cause cerebral cavernous malformation 3. Neurosurgery 57, 1008-1013.
Hashimoto, T., Wen, G., Lawton, M. T., Boudreau, N. J., Bollen, A. W., Yang, G. Y., Barbaro, N. M., Higashida, R. T., Dowd, C. F., Halbach, V. V., and Young, W. L. (2003). Abnormal expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in brain arteriovenous malformations. Stroke 34, 925-931.
Hua, J., and Muschel, R. J. (1996). Inhibition of matrix metalloproteinase 9 expression by a ribozyme blocks metastasis in a rat sarcoma model system. Cancer Res 56, 5279-5284.
Huang, C., Jacobson, K., and Schaller, M. D. (2004). MAP kinases and cell migration. J Cell Sci 117, 4619-4628.
Ishida, Y., Agata, Y., Shibahara, K., and Honjo, T. (1992). Induced expression of PD-1, a novel member of the immunoglobulin gene superfamily, upon programmed cell death. Embo J 11, 3887-3895.
Itoh, T., Tanioka, M., Matsuda, H., Nishimoto, H., Yoshioka, T., Suzuki, R., and Uehira, M. (1999). Experimental metastasis is suppressed in MMP-9-deficient mice. Clin Exp Metastasis 17, 177-181.
Jia, H. L., Ye, Q. H., Qin, L. X., Budhu, A., Forgues, M., Chen, Y., Liu, Y. K., Sun, H. C., Wang, L., Lu, H. Z., et al. (2007). Gene expression profiling reveals potential biomarkers of human hepatocellular carcinoma. Clin Cancer Res 13, 1133-1139.
Johnson, G. L., and Lapadat, R. (2002). Mitogen-activated protein kinase pathways mediated by ERK, JNK, and p38 protein kinases. Science 298, 1911-1912.
Kutuk, O., and Basaga, H. (2006). Bcl-2 protein family: implications in vascular apoptosis and atherosclerosis. Apoptosis 11, 1661-1675.
Leber, T. M., and Balkwill, F. R. (1997). Zymography: a single-step staining method for quantitation of proteolytic activity on substrate gels. Anal Biochem 249, 24-28.
Lee, C. Z., Yao, J. S., Huang, Y., Zhai, W., Liu, W., Guglielmo, B. J., Lin, E., Yang, G. Y., and Young, W. L. (2006). Dose-response effect of tetracyclines on cerebral matrix metalloproteinase-9 after vascular endothelial growth factor hyperstimulation. J Cereb Blood Flow Metab 26, 1157-1164.
Lin, J. L., Chen, H. C., Fang, H. I., Robinson, D., Kung, H. J., and Shih, H. M. (2001). MST4, a new Ste20-related kinase that mediates cell growth and transformation via modulating ERK pathway. Oncogene 20, 6559-6569.
Liquori, C. L., Berg, M. J., Squitieri, F., Ottenbacher, M., Sorlie, M., Leedom, T. P., Cannella, M., Maglione, V., Ptacek, L., Johnson, E. W., and Marchuk, D. A. (2006). Low frequency of PDCD10 mutations in a panel of CCM3 probands: potential for a fourth CCM locus. Hum Mutat 27, 118.
Ma, X., Zhao, H., Shan, J., Long, F., Chen, Y., Chen, Y., Zhang, Y., Han, X., and Ma, D. (2007). PDCD10 Interacts with Ste20-related Kinase MST4 to Promote Cell Growth and Transformation via Modulation of the ERK Pathway. Mol Biol Cell 18, 1965-1978.
Meyer, E., Vollmer, J. Y., Bovey, R., and Stamenkovic, I. (2005). Matrix metalloproteinases 9 and 10 inhibit protein kinase C-potentiated, p53-mediated apoptosis. Cancer Res 65, 4261-4272.
Otten, P., Pizzolato, G. P., Rilliet, B., and Berney, J. (1989). [131 cases of cavernous angioma (cavernomas) of the CNS, discovered by retrospective analysis of 24,535 autopsies]. Neurochirurgie 35, 82-83, 128-131.
Pozzati, E., Acciarri, N., Tognetti, F., Marliani, F., and Giangaspero, F. (1996). Growth, subsequent bleeding, and de novo appearance of cerebral cavernous angiomas. Neurosurgery 38, 662-669; discussion 669-670.
Qian, Z., Lin, C., Espinosa, R., LeBeau, M., and Rosner, M. R. (2001). Cloning and characterization of MST4, a novel Ste20-like kinase. J Biol Chem 276, 22439-22445.
Ramos-DeSimone, N., Hahn-Dantona, E., Sipley, J., Nagase, H., French, D. L., and Quigley, J. P. (1999). Activation of matrix metalloproteinase-9 (MMP-9) via a converging plasmin/stromelysin-1 cascade enhances tumor cell invasion. J Biol Chem 274, 13066-13076.
Rigamonti, D., Drayer, B. P., Johnson, P. C., Hadley, M. N., Zabramski, J., and Spetzler, R. F. (1987). The MRI appearance of cavernous malformations (angiomas). J Neurosurg 67, 518-524.
Rigamonti, D., Hadley, M. N., Drayer, B. P., Johnson, P. C., Hoenig-Rigamonti, K., Knight, J. T., and Spetzler, R. F. (1988). Cerebral cavernous malformations. Incidence and familial occurrence. N Engl J Med 319, 343-347.
Strasser, A., O'Connor, L., and Dixit, V. M. (2000). Apoptosis signaling. Annu Rev Biochem 69, 217-245.
Tanimura, S., Asato, K., Fujishiro, S. H., and Kohno, M. (2003). Specific blockade of the ERK pathway inhibits the invasiveness of tumor cells: down-regulation of matrix metalloproteinase-3/-9/-14 and CD44. Biochem Biophys Res Commun 304, 801-806.
Uhlik, M. T., Abell, A. N., Johnson, N. L., Sun, W., Cuevas, B. D., Lobel-Rice, K. E., Horne, E. A., Dell'Acqua, M. L., and Johnson, G. L. (2003). Rac-MEKK3-MKK3 scaffolding for p38 MAPK activation during hyperosmotic shock. Nat Cell Biol 5, 1104-1110.
Verlaan, D. J., Roussel, J., Laurent, S. B., Elger, C. E., Siegel, A. M., and Rouleau, G. A. (2005). CCM3 mutations are uncommon in cerebral cavernous malformations. Neurology 65, 1982-1983.
Vu, T. H., and Werb, Z. (2000). Matrix metalloproteinases: effectors of development and normal physiology. Genes Dev 14, 2123-2133.
Wang, Y. G., Liu, H. T., Zhang, Y. M., and Ma, D. L. (1999). cDNA cloning and expression of an apoptosis-related gene, human TFAR15 gene. Sci China 42, 323-329.
Westermarck, J., and Kahari, V. M. (1999). Regulation of matrix metalloproteinase expression in tumor invasion. Faseb J 13, 781-792.
Wong, M. L., Kaye, A. H., and Hovens, C. M. (2007). Targeting malignant glioma survival signalling to improve clinical outcomes. J Clin Neurosci 14, 301-308.
Yu, Q., and Stamenkovic, I. (2000). Cell surface-localized matrix metalloproteinase-9 proteolytically activates TGF-beta and promotes tumor invasion and angiogenesis. Genes Dev 14, 163-176.
Zawistowski, J. S., Serebriiskii, I. G., Lee, M. F., Golemis, E. A., and Marchuk, D. A. (2002). KRIT1 association with the integrin-binding protein ICAP-1: a new direction in the elucidation of cerebral cavernous malformations (CCM1) pathogenesis. Hum Mol Genet 11, 389-396.
Zawistowski, J. S., Stalheim, L., Uhlik, M. T., Abell, A. N., Ancrile, B. B., Johnson, G. L., and Marchuk, D. A. (2005). CCM1 and CCM2 protein interactions in cell signaling: implications for cerebral cavernous malformations pathogenesis. Hum Mol Genet 14, 2521-2531.
Zhang, J., Clatterbuck, R. E., Rigamonti, D., Chang, D. D., and Dietz, H. C. (2001). Interaction between krit1 and icap1alpha infers perturbation of integrin beta1-mediated angiogenesis in the pathogenesis of cerebral cavernous malformation. Hum Mol Genet 10, 2953-2960.