摘要
目的 建立人结肠癌SW4 8 0耐阿霉素细胞系 (SW4 8 0 /ADM) ,研究其耐药机制及逆转。方法 应用人结肠癌细胞系SW4 8 0,以递增阿霉素 (ADM)浓度的方法,体外连续培养建成一株SW4 8 0 /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 SW4 8 0 /ADM细胞与SW4 8 0细胞相比,生长缓慢,倍增时间延长,细胞周期分布发生改变;SW4 8 0 /ADM细胞较SW4 8 0的阿霉素半数抑制浓度 (IC50 )增大 1 3. 2 2倍,并对多种抗癌药物产生耐药性。SW4 8 0 /ADM细胞表面MDR的表达产物P gp,MRP和GSH/GST的表达均较SW4 8 0细胞显著升高 (P< 0. 0 1 )。SW4 8 0 /ADM细胞内阿霉素含量明显低于SW4 8 0细胞。结论 SW4 8 0 /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。
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.
出处
《中国普通外科杂志》
CAS
CSCD
2005年第3期186-189,共4页
China Journal of General Surgery
