• 1.華西醫(yī)科大學附屬第一醫(yī)院普外科(成都 610041);;
  • 2.華西醫(yī)科大學附屬第一醫(yī)院腫瘤生物治療中心;;
  • 3.華西醫(yī)科大學腫瘤研究所;;
  • 4.華西醫(yī)科大學分子生物研究室;

目的  動態(tài)觀察阿霉素(ADM)誘導(dǎo)肝癌細胞SMMC-7721耐藥性的產(chǎn)生，了解多藥耐藥相關(guān)蛋白(MRP)在其耐藥機理中的作用。
方法  分別用逐步提高培養(yǎng)基中ADM的濃度誘導(dǎo)SMMC-7721細胞和用含不同ADM濃度的培養(yǎng)基直接短期培養(yǎng)SMMC7721細胞的方法，誘導(dǎo)細胞產(chǎn)生耐藥性，繪制劑量反應(yīng)曲線，確定細胞耐藥倍數(shù)，RT-PCR法測定細胞MRP mRNA的表達水平，流式細胞儀檢測細胞內(nèi)柔紅霉素(DNR)的濃度。
結(jié)果  隨著培養(yǎng)基中ADM濃度的逐步提高，MRP mRNA的表達也逐漸增強，細胞內(nèi)DNR濃度明顯下降，SMMC-7721細胞的耐藥性逐漸增加；用含不同ADM濃度的培養(yǎng)基直接培養(yǎng)親代細胞后，雖然MRP mRNA表達明顯升高，但細胞內(nèi)DNR濃度仍維持較高水平，并且絕大部分細胞短期內(nèi)死亡。
結(jié)論  ADM可以逐步誘導(dǎo)肝癌細胞產(chǎn)生耐藥性，其機理主要是由于MRP基因過度表達，其蛋白產(chǎn)物能明顯降低細胞內(nèi)化療藥物濃度，從而細胞獲得耐藥性。

引用本文： 戴越盟,林琦遠,嚴律南,林萍,智星,雷松,屈藝. 多藥耐藥相關(guān)蛋白在阿霉素誘導(dǎo)人肝癌細胞SMMC-7721耐藥性產(chǎn)生中的作用及機理. 中國普外基礎(chǔ)與臨床雜志, 2001, 8(1): 8-11. doi: 復(fù)制

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  1. 1. Beck WT. The cell biology of multiple drug resistance[J]. Biochem Pharmac, 1987; 36(18)∶2879.
  2. 2. Barrand MA, HeppellParton AC, Wright KA, et al. A 190kilodalton protein overexpressed in nonPglycoproteincontaining multidrugresistant cells and is relationship to the MRP gene[J]. J Natl Cancer Inst, 1994; 86(2)∶110.
  3. 3. Paul S, Belinsky MG, Shen H, et al. Structure and in vitro substrate specificity of the murine multidrug resistantassociated protein [J]. Biochem, 1996; 35(42)∶13647.
  4. 4. Krishnamachary N, Center MS. The MRP gene associated with a nonpglycoprotein multidrug resistance encodes a 190KDa membrane bound glycoprotein [J]. Cancer Res, 1993; 53(16)∶3658.
  5. 5. Cole SCP, 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.
  6. 6. Endo K, Maehara Y, Kusumoto T, et al. Expression of multidrugresistanceassociated protein(MRP) and chemosensitivity in human gastric cancer [J]. Int J Cancer, 1996; 68(3)∶372.