Department of Gastroenterology, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China.
World J Gastroenterol. 2010 May 14;16(18):2291-7. doi: 10.3748/wjg.v16.i18.2291.
To establish a multidrug-resistant hepatoma cell line (SK-Hep-1), and to investigate its biological characteristics.
A highly invasive SK-Hep-1 cell line of human hepatocellular carcinoma, also known as malignant hepatoma was incubated with a high concentration of cisplatin (CDDP) to establish a CDDP-resistant cell subline (SK-Hep-1/CDDP). The 50% inhibitory dose (IC(50)) values and the resistance indexes [(IC(50) SK-Hep-1/CDDP)/(IC(50) SK-Hep-1)] for other chemotherapeutic agents and the growth curve of cells were all evaluated using cell counting kit-8 assays. The distribution of the cell cycles were detected by flow cytometry. Expression of acquired multidrug resistance P-glycoprotein (MDR1, ABCB1) and multidrug resistance-associated protein 1 (MRP1, ABCC1) was compared with that in parent cells by Western blotting and immunofluorescence combined with laser scanning confocal microscopy.
The SK-Hep-1/CDDP cells (IC(50) = 70.61 +/- 1.06 microg/mL) was 13.76 times more resistant to CDDP than the SK-Hep-1 cells (IC(50) = 5.13 +/- 0.09 microg/mL), and CDDP-resistant cells also demonstrated cross-resistance to many anti-tumor agents such as doxorubicin, 5-fluorouracil and vincristine. Similar morphologies were determined in both SK-Hep-1 and SK-Hep-1/CDDP groups. The cell cycle distribution of the SK-Hep-1/CDDP cell line exhibited a significantly increased percentage of cells in S (42.2% +/- 2.65% vs 27.91% +/- 2.16%, P < 0.01) and G2/M (20.67% +/- 5.69% vs 12.14% +/- 3.36%, P < 0.01) phases in comparison with SK-Hep-1 cells, while the percentage of cells in the G0/G1 phase decreased (37.5% +/- 5.05% vs 59.83% +/- 3.28%, P < 0.01). The levels of MDR1 and MRP1 were overexpressed in the SK-Hep-1/CDDP cells exhibiting the MDR phenotype.
Multiple drug resistance of multiple drugs in the human hepatoma cell line SK-Hep-1/CDDP was closely related to the overexpression of MDR1 and MRP1.
建立人肝癌多药耐药细胞系（SK-Hep-1），并研究其生物学特性。
用高浓度顺铂（CDDP）孵育人肝癌 SK-Hep-1 细胞系，建立 CDDP 耐药细胞亚系（SK-Hep-1/CDDP）。采用细胞计数试剂盒-8 法评估细胞对其他化疗药物的 50%抑制剂量（IC（50））值和耐药指数[（IC（50）SK-Hep-1/CDDP）/（IC（50）SK-Hep-1）]，并用流式细胞术检测细胞周期分布。Western 印迹和免疫荧光结合激光共聚焦显微镜比较亲本细胞和多药耐药相关蛋白 1（MRP1，ABCC1）表达。
SK-Hep-1/CDDP 细胞（IC（50）=70.61 +/- 1.06 microg/mL）对 CDDP 的耐药性是 SK-Hep-1 细胞（IC（50）=5.13 +/- 0.09 microg/mL）的 13.76 倍，而且 CDDP 耐药细胞对多柔比星、5-氟尿嘧啶和长春新碱等多种抗肿瘤药物也表现出交叉耐药性。SK-Hep-1 和 SK-Hep-1/CDDP 组的形态相似。SK-Hep-1/CDDP 细胞系的细胞周期分布显示 S 期（42.2% +/- 2.65% vs 27.91% +/- 2.16%，P < 0.01）和 G2/M 期（20.67% +/- 5.69% vs 12.14% +/- 3.36%，P < 0.01）的细胞比例明显增加，而 G0/G1 期的细胞比例下降（37.5% +/- 5.05% vs 59.83% +/- 3.28%，P < 0.01）。SK-Hep-1/CDDP 细胞表达 MDR1 和 MRP1，表现出多药耐药表型。
人肝癌细胞系 SK-Hep-1/CDDP 对多种药物的多药耐药与 MDR1 和 MRP1 的过度表达密切相关。
