• 華中科技大學(xué)同濟(jì)醫(yī)學(xué)院附屬同濟(jì)醫(yī)院普外科(武漢430030);

【摘要】目的 研究小分子泛素樣修飾體-1(small ubiquitin-like modifier-1，SUMO-1)在野生型p53基因誘導(dǎo)HepG2細(xì)胞凋亡中的作用。
方法 用含人野生型p53基因質(zhì)粒pcDNA3-wtp53(pwtp53)、含人雙微粒體基因2〔（human double minute gene 2 (HDM2)； 鼠同源基因?yàn)镸DM2〕質(zhì)粒pCMV-HDM1B(pMDM2)、含人SUMO-1基因質(zhì)粒pcDNA3-His6-SUMO-1(pSUMO-1)和空質(zhì)粒pcDNA3分別或同時(shí)轉(zhuǎn)染HepG2細(xì)胞，獲得各轉(zhuǎn)染細(xì)胞系，應(yīng)用Western blot檢測(cè)轉(zhuǎn)染后細(xì)胞中質(zhì)粒蛋白的表達(dá)及流式細(xì)胞技術(shù)檢測(cè)細(xì)胞凋亡比例。
結(jié)果 轉(zhuǎn)染pwtp53和pMDM2質(zhì)粒的HepG2細(xì)胞均可見p53及MDM2蛋白條帶，同時(shí)轉(zhuǎn)染pSUMO-1質(zhì)粒的細(xì)胞分別可見被SUMO-1修飾的相對(duì)分子量較大的p53和MDM2蛋白條帶，在未轉(zhuǎn)染任何質(zhì)粒、僅轉(zhuǎn)染空質(zhì)粒和pSUMO1質(zhì)粒的細(xì)胞中只檢測(cè)到少量p53蛋白表達(dá)。轉(zhuǎn)染pwtp53及pwtp53+pSUMO-1質(zhì)粒的HepG2細(xì)胞凋亡比例分別為(16.79±1.62)%和(18.15±1.36)%，轉(zhuǎn)染pwtp53+pMDM2+pSUMO-1質(zhì)粒的細(xì)胞凋亡比例為(14.06±1.84)%，轉(zhuǎn)染pwtp53+pMDM2質(zhì)粒的細(xì)胞凋亡比例則下降至(5.17±1.23)%，與前三者比較差異有顯著性意義(PH＜0.01)； 其他轉(zhuǎn)染系細(xì)胞中的細(xì)胞凋亡比例均≤2%，差異無顯著性意義。
結(jié)論 SUMO-1通過與p53蛋白的結(jié)合或翻譯后修飾，抑制MDM2等癌基因蛋白對(duì)p53蛋白的降解，可增強(qiáng)p53抑癌基因誘導(dǎo)的細(xì)胞凋亡。

引用本文： 盧星榕,易繼林. SUMO-1對(duì)p53基因誘導(dǎo)HepG2細(xì)胞凋亡的增強(qiáng)作用. 中國普外基礎(chǔ)與臨床雜志, 2005, 12(4): 371-374. doi: 復(fù)制

1. Ko LJ, Prives C. p53: puzzle and paradigm [Ｊ]. Genes Dev, 1996; 10(9)∶1054.
2. Rosenfeld MR, Meneses P, Dalmau J, et al. Gene transfer of wildtype p53 results in restoration of tumorsuppressor function in a medulloblastoma cell line [Ｊ]. Neurology, 1995; 45(8)∶1533.
3. Honda R, Tanaka H, Yasuda H. Oncoprotein MDM2 is a ubiquitin ligase E3 for tumor suppressor p53 [Ｊ]. FEBS Lett, 1997; 420(1)∶25.
4. Haupt Y, Maya R, Kazaz A, et al. Mdm2 promotes the rapid degradation of p53 [Ｊ]. Nature, 1997; 387(6630)∶296.
5. Saitoh H,Hinchey J. Functional heterogeneity of small ubiquitinrelated protein modifiers SUMO1 versus SUMO2/3 [Ｊ]. J Biol Chem, 2000; 275(9)∶6252.
6. Bayer P, Arndt A, Metzger S, et al. Structure determination of the small ubiquitinrelated modifier SUMO1 [Ｊ]. J Mol Biol, 1998; 280(2)∶275.
7. Muller S, Hoege C, Pyrowolakis G, et al. SUMO, ubiquitin’s mysterious cousin [Ｊ]. Nat Rev Mol Cell Biol, 2001; 2(3)∶202.
8. Melchior F. SUMO——nonclassical ubiquitin [Ｊ]. Annu Rev Cell Dev Biol, 2000; 16∶591.
9. Rodriguez MS, Desterro JM, Lain S, et al. SUMO1 modification activates the transcriptional response of p53 [Ｊ]. EMBO J, 1999; 18(22)∶6455.
10. Gostissa M, Hengstermann A, Fogal V, et al. Activation of p53 by conjugation to the ubiquitinlike protein SUMO1 [Ｊ]. EMBO J, 1999; 18(22)∶6462.
11. Sambrook J, Ftitsch EF, Mauitis T. Molecular cloning.A laboratory manual [M]. 2nd. New York: Cold Spring Harbor Laboratory Press, 1988∶362-371,792-793.
12. Zheleva DI, Lane DP, Fischer PM. The p53Mdm2 pathway: targets for the development of new anticancer therapeutics [Ｊ]. Mini Rev Med Chem, 2003; 3(3)∶257.
13. Kurepa J, Walker JM, Smalle J, et al. The small ubiquitinlike modifier (SUMO) protein modification system in Arabidopsis. Accumulation of SUMO1 and 2 conjugates is increased by stress [Ｊ]. J Biol Chem, 2003; 278(9)∶6862.
  1. 1. Ko LJ, Prives C. p53: puzzle and paradigm [Ｊ]. Genes Dev, 1996; 10(9)∶1054.
  2. 2. Rosenfeld MR, Meneses P, Dalmau J, et al. Gene transfer of wildtype p53 results in restoration of tumorsuppressor function in a medulloblastoma cell line [Ｊ]. Neurology, 1995; 45(8)∶1533.
  3. 3. Honda R, Tanaka H, Yasuda H. Oncoprotein MDM2 is a ubiquitin ligase E3 for tumor suppressor p53 [Ｊ]. FEBS Lett, 1997; 420(1)∶25.
  4. 4. Haupt Y, Maya R, Kazaz A, et al. Mdm2 promotes the rapid degradation of p53 [Ｊ]. Nature, 1997; 387(6630)∶296.
  5. 5. Saitoh H,Hinchey J. Functional heterogeneity of small ubiquitinrelated protein modifiers SUMO1 versus SUMO2/3 [Ｊ]. J Biol Chem, 2000; 275(9)∶6252.
  6. 6. Bayer P, Arndt A, Metzger S, et al. Structure determination of the small ubiquitinrelated modifier SUMO1 [Ｊ]. J Mol Biol, 1998; 280(2)∶275.
  7. 7. Muller S, Hoege C, Pyrowolakis G, et al. SUMO, ubiquitin’s mysterious cousin [Ｊ]. Nat Rev Mol Cell Biol, 2001; 2(3)∶202.
  8. 8. Melchior F. SUMO——nonclassical ubiquitin [Ｊ]. Annu Rev Cell Dev Biol, 2000; 16∶591.
  9. 9. Rodriguez MS, Desterro JM, Lain S, et al. SUMO1 modification activates the transcriptional response of p53 [Ｊ]. EMBO J, 1999; 18(22)∶6455.
  10. 10. Gostissa M, Hengstermann A, Fogal V, et al. Activation of p53 by conjugation to the ubiquitinlike protein SUMO1 [Ｊ]. EMBO J, 1999; 18(22)∶6462.
  11. 11. Sambrook J, Ftitsch EF, Mauitis T. Molecular cloning.A laboratory manual [M]. 2nd. New York: Cold Spring Harbor Laboratory Press, 1988∶362-371,792-793.
  12. 12. Zheleva DI, Lane DP, Fischer PM. The p53Mdm2 pathway: targets for the development of new anticancer therapeutics [Ｊ]. Mini Rev Med Chem, 2003; 3(3)∶257.
  13. 13. Kurepa J, Walker JM, Smalle J, et al. The small ubiquitinlike modifier (SUMO) protein modification system in Arabidopsis. Accumulation of SUMO1 and 2 conjugates is increased by stress [Ｊ]. J Biol Chem, 2003; 278(9)∶6862.