人结肠癌耐药细胞系SW480/ADM的建立及其生物学特性
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黄柏英

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Establishment of human colon carcinoma multidrug-resistant SW480/ADM cell line and study of its biological characteristics
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    摘要:

    目的:建立人结肠癌SW480耐阿霉素细胞系(SW480/ADM) ,研究其耐药机制及逆转。方法:应用人结肠癌细胞系SW480,以递增阿霉素(ADM)浓度的方法,体外连续培养建成一株SW480/ADM。观察该细胞生长规律。用MTT比色法检测该耐药细胞系的多药耐药性;以流式细胞术检测该耐药细胞的细胞周期分布、细胞表面多药耐药基因(MDR)的表达产物P-糖蛋白(P-gp) 、多药耐药相关蛋白(MRP)及谷胱甘肽硫转移系统(GSH/GST)的表达;用高效液相色谱仪检测耐药细胞内阿霉素含量。 结果:SW480/ADM细胞与SW480细胞相比,生长缓慢,倍增时间延长,细胞周期分布发生改变;SW480/ADM细胞较SW480的阿霉素半数抑制浓度(IC 50)增大13.22倍,并对多种抗癌药物产生耐药性。SW480/ADM细胞表面MDR的表达产物P-gp,MRP和GSH/GST的表达均较SW480细胞显著升高(P<0.01)。SW480/ADM细胞内阿霉素含量明显低于SW480细胞。 结论:SW480/ADM细胞对阿霉素的耐药是获得性的,并呈多药耐药性特征,可用于对人结肠癌耐药、逆转和MDR机制等方面的研究。

    Abstract:

    Abstract:Objective:To establish an adriamycin resistant human colon cancer cell line SW480 (SW480/ADM),and study their drug resistance mechanism and reversal.Methods:An ADM-resistant human colon carcinoma cell line SW480/ADM was induced by continuously exposing human colon carcinoma cell line SW480 to gradually increasing doses of Adriamycin(ADM). The multidrug resistance of SW480/ADM was evaluated by MTT assay. The distribution of its cell cycle, the expressions of P-gp, multidrug resistance-associated protein(MRP) and GSH/GST were detected by flow cytometry. ADM content in the SW480/ADM was detected by high-performance liquid chromatography(HPLC). Results:Compared with parental cell line SW480, the SW480/ADM cell line had a slower growth rate and longer doubling time,and distribution of its cell cycle had changed. SW480/ADM was resistant to many anti-tumor agents. IC 50 of ADM of SW480/ADM cells was 13.22 times higher than that of parental cell line SW480, and the expressions of P-gp,MRP and GSH/GST were enhanced significantly(P<0.01). ADM content of the SW480/ADM cells was markedly lower than that of SW480 cells. Conclusions:The resistance of SW480/ADM cells to ADM was acquired and had the characteristic of multidrug resistance,and can be used for the study of drug-resistance, reversal and MDR mechanism of human colon carcinoma.

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黄柏英,祝和成,顾焕华.人结肠癌耐药细胞系SW480/ADM的建立及其生物学特性[J].中国普通外科杂志,2005,14(3):7-.
DOI:10.7659/j. issn.1005-6947.2005.03.007

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  • 在线发布日期: 2005-03-25