• 昆明醫(yī)學(xué)院第二附屬醫(yī)院肝膽外科（昆明650101）;

目的 　研究pcDNA3/AFP/TK/Angio融合基因?qū)θ烁伟┘?xì)胞系SMMC-7721裸鼠移植瘤模型的靶向性治療作用。
方法 　建立人原發(fā)性肝癌裸鼠皮下移植瘤模型，將荷瘤裸鼠隨機分成腫瘤對照組、空質(zhì)粒組、丙氧鳥苷(GCV）組、pcDNA3/TK/Angio組及pcDNA3/AFP/TK/Angio組5組。于瘤體內(nèi)分別直接注射不同的質(zhì)粒，同時于裸鼠腹腔內(nèi)注射GCV，觀察不同時段皮下腫瘤的生長情況并做病理學(xué)檢查，免疫組化法檢測腫瘤微血管密度（MVD）以及血管內(nèi)皮細(xì)胞生長因子（VEGF）的表達(dá)量，原位末端標(biāo)記(TUNEL)法檢測細(xì)胞原位凋亡。放免法檢測裸鼠血清甲胎蛋白（AFP）的變化，透射電鏡觀察腫瘤細(xì)胞超微結(jié)構(gòu)的變化。
結(jié)果 　裸鼠皮下成瘤率100%； pcDNA3/TK/Angio組及pcDNA3/AFP/TK/Angio組的腫瘤體積、血清AFP含量、腫瘤MVD和VEGF表達(dá)強度均明顯低于對照組、空質(zhì)粒組和GCV組（P＜0.05），細(xì)胞凋亡指數(shù)都明顯高于后3組（P＜0.05），可見較多的凋亡細(xì)胞。而pcDNA3/AFP/TK/Angio組的腫瘤體積、AFP、MVD及VEGF表達(dá)強度又明顯低于pcDNA3/TK/Angio組（P＜0.05），凋亡指數(shù)高于后者（P＜0.05）。
結(jié)論 　pcDNA3/AFP/TK/Angio融合基因系統(tǒng)可顯著抑制腫瘤的生長，有望成為治療原發(fā)性肝癌的新型生物制劑之一。

引用本文： 盛勤松,王琳,任鋒,盧永剛,鄒浩,肖曙峰,唐波,黃潔,張捷. pcDNA3/AFP/TK/Angio融合基因靶向性治療人原發(fā)性肝癌的實驗研究. 中國普外基礎(chǔ)與臨床雜志, 2007, 14(4): 403-408. doi: 復(fù)制

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6. 　李梅生, 梁力建, 黃潔夫, 等. HSV-tk/CD融合基因殺傷人膽管癌細(xì)胞QBC939的體內(nèi)實驗 [Ｊ]　. 中國普外基礎(chǔ)與臨床雜志, 2005; 12（6）∶581.
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8. 　Tai KF, Chen PJ, Chen DS, et al. Concurrent delivery of GM-CSF and endostatin genes by a single adenoviral vector provides a synergistic effect on the treatment of orthotopic liver tumors [Ｊ]　. J Gene Med, 2003; 5(5)∶386.
9. 　Shi M, Wang FS, Wu ZZ. Synergetic anticancer effect of combined quercetin and recombinant adenoviral vector expressing human wild-type p53, GM-CSF and B7-1 genes on hepatocellular carcinoma cells in vitro [Ｊ]　. World J Gastroenterol, 2003; 9(1)∶73.
10. 　Sung MW, Yeh HC, Thung SN, et al. Intratumoral adenovirus-mediated suicide gene transfer for hepatic metastases from colorectal adenocarcinoma: results of a phase Ⅰ clinical trial [Ｊ]　. Mol Ther, 2001; 4(3)∶182.
11. 　Xu R, Sun X, Tse LY, et al. Long-term expression of angiostatin suppresses metastatic liver cancer in mice [Ｊ]　. Hepatology, 2003; 37(6)∶1451.
12. 　李曉, 王琳, 郭永章, 等. 血管生成抑制素基因?qū)β闶笕烁伟┮浦擦龅闹委熥饔?[Ｊ]　. 中華實驗外科雜志, 2004; 21(7)∶875　.
13. 　Shi YJ, Gong JP, Liu CA, et al. Construction of a targeting adenoviral vector carrying AFP promoter for expressing EGFP gene in AFP-producing hepatocarcinoma cell [Ｊ]　. World J Gastroenterol, 2004; 10(2)∶186.
14. 　Huang X, Zhang W, Wakimoto H, et al. Adenovirus-mediated tissue-specific cytosine deaminase gene therapy for human hepatocellular carcinoma with different AFP expression levels [Ｊ]　. J Exp Ther Oncol, 2002; 2(2)∶100.
15. 　Ohguchi S, Nakatsukasa H, Higashi T, et al. Expression of alpha-fetoprotein and albumin genes in human hepatocellular carcinomas: limitations in the application of the genes for targeting human hepatocellular carcinoma in gene therapy [Ｊ]　. Hepatology, 1998; 27(2)∶599.
  1. 1. 　Choi EA, Lei H, Maron DJ, et al. Combined 5-fluorouracil/systemic interferon-beta gene therapy results in long-term survival in mice with established colorectal liver metastases [Ｊ]　. Clin Cancer Res, 2004; 10(4)∶1535.
  2. 2. 　Hwang KS, Cho WK, Yoo J, et al. Adenovirus-mediated interleukin-12 gene transfer combined with cytosine deaminase followed by 5-fluorocytosine treatment exerts potent antitumor activity in Renca tumor-bearing mice [Ｊ]　. BMC Cancer, 2005; 5(1)∶51.
  3. 3. 　You TG, Wang HS, Yang JH, et al. Transfection of IL-2 and/or IL-12 genes into spleen in treatment of rat liver cancer [Ｊ]　. World J Gastroenterol, 2004; 10(15)∶2190.
  4. 4. 　Chang CJ, Chen YH, Huang KW, et al. Combined GM-CSF and IL-12 gene therapy synergistically suppresses the growth of orthotopic liver tumors [Ｊ]　. Hepatology, 2007; 45(3)∶746.
  5. 5. 　黃畦, 陳大瑜, 付向?qū)? 等. 雙自殺基因重組腺病毒對肺癌細(xì)胞的殺傷作用 [Ｊ]　. 中華實驗外科雜志, 2006; 23(4)∶579.
  6. 6. 　李梅生, 梁力建, 黃潔夫, 等. HSV-tk/CD融合基因殺傷人膽管癌細(xì)胞QBC939的體內(nèi)實驗 [Ｊ]　. 中國普外基礎(chǔ)與臨床雜志, 2005; 12（6）∶581.
  7. 7. 　Li PY, Lin JS, Feng ZH, et al. Combined gene therapy of endostatin and interleukin 12 with polyvinylpyrrolidone induces a potent antitumor effect on hepatoma [Ｊ]　. World J Gastroenterol, 2004; 10(15)∶2195.
  8. 8. 　Tai KF, Chen PJ, Chen DS, et al. Concurrent delivery of GM-CSF and endostatin genes by a single adenoviral vector provides a synergistic effect on the treatment of orthotopic liver tumors [Ｊ]　. J Gene Med, 2003; 5(5)∶386.
  9. 9. 　Shi M, Wang FS, Wu ZZ. Synergetic anticancer effect of combined quercetin and recombinant adenoviral vector expressing human wild-type p53, GM-CSF and B7-1 genes on hepatocellular carcinoma cells in vitro [Ｊ]　. World J Gastroenterol, 2003; 9(1)∶73.
  10. 10. 　Sung MW, Yeh HC, Thung SN, et al. Intratumoral adenovirus-mediated suicide gene transfer for hepatic metastases from colorectal adenocarcinoma: results of a phase Ⅰ clinical trial [Ｊ]　. Mol Ther, 2001; 4(3)∶182.
  11. 11. 　Xu R, Sun X, Tse LY, et al. Long-term expression of angiostatin suppresses metastatic liver cancer in mice [Ｊ]　. Hepatology, 2003; 37(6)∶1451.
  12. 12. 　李曉, 王琳, 郭永章, 等. 血管生成抑制素基因?qū)β闶笕烁伟┮浦擦龅闹委熥饔?[Ｊ]　. 中華實驗外科雜志, 2004; 21(7)∶875　.
  13. 13. 　Shi YJ, Gong JP, Liu CA, et al. Construction of a targeting adenoviral vector carrying AFP promoter for expressing EGFP gene in AFP-producing hepatocarcinoma cell [Ｊ]　. World J Gastroenterol, 2004; 10(2)∶186.
  14. 14. 　Huang X, Zhang W, Wakimoto H, et al. Adenovirus-mediated tissue-specific cytosine deaminase gene therapy for human hepatocellular carcinoma with different AFP expression levels [Ｊ]　. J Exp Ther Oncol, 2002; 2(2)∶100.
  15. 15. 　Ohguchi S, Nakatsukasa H, Higashi T, et al. Expression of alpha-fetoprotein and albumin genes in human hepatocellular carcinomas: limitations in the application of the genes for targeting human hepatocellular carcinoma in gene therapy [Ｊ]　. Hepatology, 1998; 27(2)∶599.