• 1.四川大學華西醫(yī)院普外科（成都610041）;;
  • 2.成都地奧制約集團有限公司新藥部（成都610041）;;
  • 通迅作者： 嚴律南;

目的  構(gòu)建攜帶反義二型基質(zhì)金屬蛋白酶（MMP2）基因的重組腺病毒。
方法  從新鮮肝癌組織中提取總RNA,用RT-PCR法合成MMP2 cDNA序列中5′端轉(zhuǎn)錄起始位點附近長約500 bp的基因片段，將此片段反義克隆到腺病毒載體(AdEasy)系統(tǒng)的多克隆位點，經(jīng)轉(zhuǎn)染293細胞生成攜帶反義MMP2基因的重組腺病毒AdMMP2AS。
結(jié)果  成功構(gòu)建并包裝攜帶反義MMP2基因片段的重組腺病毒Ad-MMP2AS，病毒滴度達1×108/ml。
結(jié)論  構(gòu)建的重組腺病毒Ad-MMP2AS可望能有效地將反義MMP2基因片段導入人肝癌細胞株，為進一步研究肝癌浸潤和轉(zhuǎn)移機理以及探討抑制肝癌浸潤和轉(zhuǎn)移的方法提供實驗基礎(chǔ)。

引用本文： 張明滿,李德華,茍興華,嚴律南,韓蕾,趙蘭英,胡海洋. 攜反義二型基質(zhì)金屬蛋白酶基因的重組腺病毒的構(gòu)建. 中國普外基礎(chǔ)與臨床雜志, 2004, 11(5): 428-432. doi: 復制

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  2. 2. Niu Q, Tang Z, Ma Z, et al. Relationship between serum matrix metalloproteinase2 and metastasis and recurrence following radical hepatic resection in hepatocellular carcinoma [Ｊ]. Zhonghua Gan Zang Bing Za Zhi, 2001; 9（Suppl）∶58.
  3. 3. 王愛民，江龍安，梁保忠，等. 原發(fā)性肝癌組織中明膠酶A基因表達增強 [Ｊ]. 中華普通外科雜志， 2001； 15（9）∶598.
  4. 4. He TC, Zhou S, da Costa LT, et al. A simplified system for generating recombinant adenoviruses [Ｊ]. Proc Natl Acad Sci USA， 1998; 95(5)∶ 2509.
  5. 5. Curran S, Murray GI. Matrix metalloproteinases in tumour invasion and metastasis [Ｊ]. J Pathol， 1999; 189(3)∶ 300.
  6. 6. Liotta LA, Tryggvason K, Garbisa S, et al. Metastatic potential correlates with enzymatic degradation of basement membrane collagen [Ｊ]. Nature， 1980; 284(5751)∶ 67.
  7. 7. Deryugina EI, Luo GX, Reisfeld RA, et al. Tumor cell invasion through matrigel is regulated by activated matrix metalloproteinase2 [Ｊ]. Anticancer Res， 1997; 17(5A)∶ 3201.
  8. 8. Sang QX. Complex role of matrix metalloproteinases in angiogenesis [Ｊ]. Cell Res， 1998; 8(3)∶ 171.
  9. 9. McKenna GJ, Chen Y, Smith RM, et al. A role for matrix metalloproteinases and tumor host interaction in hepatocellular carcinomas [Ｊ]. Am J Surg， 2002; 183(5)∶ 588.
  10. 10. Carter G, Lemoine NR. Antisense technology for cancer therapy: does it make sense? [Ｊ]. Br J Cancer, 1993; 67(5)∶ 869.
  11. 11. Lakka SS, Rajan M, Gondi C, et al. Adenovirusmediated expression of antisense MMP9 in glioma cells inhibits tumor growth and invasion [Ｊ]. Oncogene, 2002; 21(52)∶ 8011.