• 四川大學(xué)華西醫(yī)院神經(jīng)內(nèi)科（成都，610041）;

【摘要】　目的　 探討佛波酯激活的蛋白激酶C與扭轉(zhuǎn)蛋白A在亞細(xì)胞成分中的表達(dá)之間的關(guān)系。　方法　 采用免疫熒光法觀察扭轉(zhuǎn)蛋白A在原代培養(yǎng)的神經(jīng)元和小鼠胚胎成纖維細(xì)胞（NIH 3T3細(xì)胞）中的分布。運(yùn)用蛋白質(zhì)印跡法分析蛋白激酶C和扭轉(zhuǎn)蛋白A在細(xì)亞細(xì)胞成分中的表達(dá)。　結(jié)果　 扭轉(zhuǎn)蛋白A在NIH 3T3細(xì)胞中的表達(dá)類似于神經(jīng)元。扭轉(zhuǎn)蛋白A在細(xì)胞質(zhì)溶質(zhì)、膜成分中均有分布。佛波酯活化蛋白激酶C后并不引起扭轉(zhuǎn)蛋白A在細(xì)胞質(zhì)成分和膜成分中表達(dá)含量的變化。　結(jié)論　 扭轉(zhuǎn)蛋白A可能是膜相關(guān)蛋白，細(xì)胞氧化應(yīng)激中扭轉(zhuǎn)蛋白A表達(dá)上調(diào)和重分布變化不是由佛波酯誘導(dǎo)的蛋白激酶C活化途徑來實(shí)現(xiàn)的。鑒于扭轉(zhuǎn)蛋白A表達(dá)上調(diào)具有潛在的治療原發(fā)性早發(fā)扭轉(zhuǎn)性肌張力障礙的前景，影響其分布和表達(dá)的分子機(jī)制需要進(jìn)一步研究。
【Abstract】　Objective　 To investigate the relationship between the phorbol 12-myristate 13-acetate (PMA) activated protein kinase C (PKC) and the subcellular expression of TorsinA protein.　Methods　 The expression of TorsinA in the primary cultured neurons and the NIH 3T3 cells was detected by immunofluorescence. The expression of PKC and TorsinA in subcellular fraction was analyzed by the western blotting.　Results　 The expression pattern of TorsinA in NIH 3T3 cells was similar to neuron. PMA, an activator of PKC, did not promote the up-expression of TorsinA or redistribution in the subcellular fraction of NIH 3T3 cells.　Conclusions　 TorsinA may be a membrane-associated protein. The up-regulation and redistribution of TorsinA is not caused by the pathway of the PMA activating PKC after cells insulted by oxidative stress. We should pay more attention on the mechanisms of the expression of TorsinA protein for the potential therapies to early-onset primary torsion dystonia (DYT1).

引用本文： 胡小輝,鄭珍珍,盛麗琴,潘平雷,宋偉,黃睿,商慧芳. 佛波酯誘導(dǎo)的蛋白激酶C活化對(duì)扭轉(zhuǎn)蛋白A亞細(xì)胞分布的影響. 華西醫(yī)學(xué), 2011, 26(3): 321-324. doi: 復(fù)制

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2. 　Zorzi G, Zibordi F, Garavaglia B, et al. Early onset primary dystonia[J]. Eur J Paediatr Neuro, 2009, 13(6): 488-492.
3. 　Hu XH, Chen XP, Huang R, et al. Role of DYT1 gene in early-onset primary torsion dystonia[J]. NRR, 2010, 5(18): 1429-1434.
4. 　Chen XP, Hu XH, Wu SH, et al. RNA interference-mediated inhibition of wild-type Torsin A expression increases apoptosis caused by oxidative stress in cultured cells[J]. Neurochem Res, 2010, 35(8): 1214-1223.
5. 　Hewett J, Ziefer P, Bergeron D, et al. TorsinA in PC12 cells: localization in the endoplasmic reticulum and response to stress[J]. J Neurosci Res, 2003, 72(2): 158-168.
6. 　Zhao Y, Xiao J, Ueda M, et al. Glial elements contribute to stress-induced torsinA expression in the CNS and peripheral nervous system[J]. Neuroscience, 2008, 155(2): 439-453.
7. 　Ozelius LJ, Hewett JW, Page CE, et al. The early-onset torsion dystonia gene (DYT1) encodes an ATP-binding protein[J]. Nat Genet, 1997, 17(1): 40-48.
8. 　Nery FC, Zeng J, Niland BP, et al. TorsinA binds the KASH domain of nesprins and participates in linkage between nuclear envelope and cytoskeleton[J]. J Cell Sci, 2008, 121(Pt 20): 3476-3486.
9. 　Kazi JU, Soh JW. Isoform-specific translocation of PKC isoforms in NIH3T3 cells by TPA[J]. Biochem Bioph Res Co, 2007, 364(2): 231-237.
10. 　Quest AF, Bell RM. The regulatory region of protein kinase C gamma. Studies of phorbol ester binding to individual and combined functional segments expressed as glutathione S-transferase fusion proteins indicate a complex mechanism of regulation by phospholipids, phorbol esters, and divalent cations[J]. J Biol Chem, 1994, 269(31): 20000-20012.〖ZK)〗.
11. 　Gerace L. TorsinA and torsion dystonia: unraveling the architecture of the nuclear envelope[J]. Proc Natl Acad Sci USA, 2004, 101(24): 8839-8840.
12. 　Gopalakrishna R, Jaken S. Protein kinase C signaling and oxidative stress[J]. Free Radical Bio Med, 2000, 28(9): 1349-1361.
13. 　Hewett J, Gonzalez-Agosti C, Slater D, et al. Mutant torsinA, responsible for early-onset torsion dystonia, forms membrane inclusions in cultured neural cells[J]. Hum Mol Genet, 2000, 9(9): 1403-1413.
14. 　de Carvalho Aguiar PM, Ozelius LJ. Classification and genetics of dystonia[J]. Lancet Neurol, 2002, 1(5): 316-325.
15. 　Lee SH, Dominguez R. Regulation of actin cytoskeleton dynamics in cells[J]. Mol Cells, 2010, 29(4): 311-325.
  1. 1. [10　Hsieh CH, Rau CS, Jeng SF, et al. Identification of the potential target genes of microRNA-146a induced by PMA treatment in human microvascular endothelial cells[J]. Exp Cell Res, 2010, 316(7): 1119-1126.
  2. 2. 　Zorzi G, Zibordi F, Garavaglia B, et al. Early onset primary dystonia[J]. Eur J Paediatr Neuro, 2009, 13(6): 488-492.
  3. 3. 　Hu XH, Chen XP, Huang R, et al. Role of DYT1 gene in early-onset primary torsion dystonia[J]. NRR, 2010, 5(18): 1429-1434.
  4. 4. 　Chen XP, Hu XH, Wu SH, et al. RNA interference-mediated inhibition of wild-type Torsin A expression increases apoptosis caused by oxidative stress in cultured cells[J]. Neurochem Res, 2010, 35(8): 1214-1223.
  5. 5. 　Hewett J, Ziefer P, Bergeron D, et al. TorsinA in PC12 cells: localization in the endoplasmic reticulum and response to stress[J]. J Neurosci Res, 2003, 72(2): 158-168.
  6. 6. 　Zhao Y, Xiao J, Ueda M, et al. Glial elements contribute to stress-induced torsinA expression in the CNS and peripheral nervous system[J]. Neuroscience, 2008, 155(2): 439-453.
  7. 7. 　Ozelius LJ, Hewett JW, Page CE, et al. The early-onset torsion dystonia gene (DYT1) encodes an ATP-binding protein[J]. Nat Genet, 1997, 17(1): 40-48.
  8. 8. 　Nery FC, Zeng J, Niland BP, et al. TorsinA binds the KASH domain of nesprins and participates in linkage between nuclear envelope and cytoskeleton[J]. J Cell Sci, 2008, 121(Pt 20): 3476-3486.
  9. 9. 　Kazi JU, Soh JW. Isoform-specific translocation of PKC isoforms in NIH3T3 cells by TPA[J]. Biochem Bioph Res Co, 2007, 364(2): 231-237.
  10. 10. 　Quest AF, Bell RM. The regulatory region of protein kinase C gamma. Studies of phorbol ester binding to individual and combined functional segments expressed as glutathione S-transferase fusion proteins indicate a complex mechanism of regulation by phospholipids, phorbol esters, and divalent cations[J]. J Biol Chem, 1994, 269(31): 20000-20012.〖ZK)〗.
  11. 11. 　Gerace L. TorsinA and torsion dystonia: unraveling the architecture of the nuclear envelope[J]. Proc Natl Acad Sci USA, 2004, 101(24): 8839-8840.
  12. 12. 　Gopalakrishna R, Jaken S. Protein kinase C signaling and oxidative stress[J]. Free Radical Bio Med, 2000, 28(9): 1349-1361.
  13. 13. 　Hewett J, Gonzalez-Agosti C, Slater D, et al. Mutant torsinA, responsible for early-onset torsion dystonia, forms membrane inclusions in cultured neural cells[J]. Hum Mol Genet, 2000, 9(9): 1403-1413.
  14. 14. 　de Carvalho Aguiar PM, Ozelius LJ. Classification and genetics of dystonia[J]. Lancet Neurol, 2002, 1(5): 316-325.
  15. 15. 　Lee SH, Dominguez R. Regulation of actin cytoskeleton dynamics in cells[J]. Mol Cells, 2010, 29(4): 311-325.