• 1.四川大學華西醫(yī)院普外科（成都610041）;;
  • 　2.成都地奧制藥集團有限公司基因工程藥物研究室;

目的構(gòu)建攜帶反義多藥耐藥相關(guān)蛋白(MRP)的重組腺病毒并轉(zhuǎn)染人肝癌耐藥細胞株，為肝癌耐藥的基因治療提供實驗研究基礎(chǔ)。 方法將MRP基因片段反向克隆到腺病毒載體質(zhì)粒pAdTrackCMV上，與骨架質(zhì)粒在細菌體內(nèi)進行同源重組，經(jīng)293細胞包裝、擴增后得到攜帶反義MRP的重組腺病毒AdEasyGFPASmrp，再用其轉(zhuǎn)染人肝癌耐藥細胞株SMMC7721/ADM。結(jié)果成功地構(gòu)建了攜帶反義MRP的重組腺病毒，病毒滴度為2.5×109 efu/ml，多重感染復數(shù)為100時對SMMC7721/ADM細胞株轉(zhuǎn)導效率可達90%以上。結(jié)論構(gòu)建的重組腺病毒AdEasyGFPASmrp可望有效地將反義MRP導入人肝癌耐藥細胞株，為肝癌耐藥機理及其逆轉(zhuǎn)方式的研究提供實驗基礎(chǔ)。

引用本文： 陳琳,茍興華,嚴律南,趙永恒,韓蕾,李德華,胡海洋,趙蘭英. 攜帶反義多藥耐藥相關(guān)蛋白的重組腺病毒的構(gòu)建及其應用的初步研究△. 中國普外基礎(chǔ)與臨床雜志, 2003, 10(2): 117-120. doi: 復制

1. Cole SP, Bhardwaj G, Gerlach JH, et al. Overexpression of a transporter gene in a multidrugresistant human lung cancer cell line　[Ｊ]. Science， 1992; 258(5088)∶1650.
2. Hipfner DR,Deeley RG,Cole SP.Structural, mechanistic and clinical aspects of MRP1　[Ｊ]. Biochim Biophys Acta,1999; 1461(2)∶359.
3. Roelofsen H, Vos TA, Schippers IJ, et al. Increased levels of the multidrug resistance protein in lateral membranes of proliferating hepatocytederived cells [Ｊ]. Gastroenterology, 1997; 112(2)∶ 511.
4. 戴越盟，林琦遠，嚴律南,等. 多藥耐藥相關(guān)蛋白在阿霉素誘導人肝癌細胞SMMC7721耐藥性產(chǎn)生中的作用及機理　[Ｊ]. 中國普外基礎(chǔ)與臨床雜志，2001； 8（1）∶8.
5. Carter G, Lemoine NR. Antisense technology for cancer therapy: does it make sense?　[Ｊ]. Br J Cancer, 1993; 67(5)∶869.
6. 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.
7. Inoue H, Nojima H, Okayama H. High efficiency transformation of Escherichia coli with plasmids　[Ｊ]. Gene, 1990; 96(1)∶23.
8. 司徒振強，吳軍正主編. 細胞培養(yǎng)　[M]. 第1版.西安： 世界圖書出版公司，1996∶186～187.
9. Itsubo M,Ishikawa T,Toda G,et al.Immunohistochemic study of expression and cellular localization of the multidrug resistance gene product Pglycoprotein in primary liver carcinoma　[Ｊ].Cancer,1994; 73(2)∶ 298.
10. Lai EC, Choi TK, Cheng CH, et al. Doxorubicin for unresectable hepatocellular carcinoma. A prospective study on the addition of verapami　[Ｊ]. Cancer, 1990; 66(8)∶1685.
11. Bradshaw DM, Arceci RJ. Clinical relevance of transmembrane drug efflux as a mechanism of multidrug resistance　[Ｊ]. J Clin Oncol, 1998; 16(11)∶3674.
  1. 1. Cole SP, Bhardwaj G, Gerlach JH, et al. Overexpression of a transporter gene in a multidrugresistant human lung cancer cell line　[Ｊ]. Science， 1992; 258(5088)∶1650.
  2. 2. Hipfner DR,Deeley RG,Cole SP.Structural, mechanistic and clinical aspects of MRP1　[Ｊ]. Biochim Biophys Acta,1999; 1461(2)∶359.
  3. 3. Roelofsen H, Vos TA, Schippers IJ, et al. Increased levels of the multidrug resistance protein in lateral membranes of proliferating hepatocytederived cells [Ｊ]. Gastroenterology, 1997; 112(2)∶ 511.
  4. 4. 戴越盟，林琦遠，嚴律南,等. 多藥耐藥相關(guān)蛋白在阿霉素誘導人肝癌細胞SMMC7721耐藥性產(chǎn)生中的作用及機理　[Ｊ]. 中國普外基礎(chǔ)與臨床雜志，2001； 8（1）∶8.
  5. 5. Carter G, Lemoine NR. Antisense technology for cancer therapy: does it make sense?　[Ｊ]. Br J Cancer, 1993; 67(5)∶869.
  6. 6. 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.
  7. 7. Inoue H, Nojima H, Okayama H. High efficiency transformation of Escherichia coli with plasmids　[Ｊ]. Gene, 1990; 96(1)∶23.
  8. 8. 司徒振強，吳軍正主編. 細胞培養(yǎng)　[M]. 第1版.西安： 世界圖書出版公司，1996∶186～187.
  9. 9. Itsubo M,Ishikawa T,Toda G,et al.Immunohistochemic study of expression and cellular localization of the multidrug resistance gene product Pglycoprotein in primary liver carcinoma　[Ｊ].Cancer,1994; 73(2)∶ 298.
  10. 10. Lai EC, Choi TK, Cheng CH, et al. Doxorubicin for unresectable hepatocellular carcinoma. A prospective study on the addition of verapami　[Ｊ]. Cancer, 1990; 66(8)∶1685.
  11. 11. Bradshaw DM, Arceci RJ. Clinical relevance of transmembrane drug efflux as a mechanism of multidrug resistance　[Ｊ]. J Clin Oncol, 1998; 16(11)∶3674.