Brief Article
Copyright ©2010 Baishideng. All rights reserved
World J Gastroenterol. May 14, 2010; 16(18): 2291-2297
Published online May 14, 2010. doi: 10.3748/wjg.v16.i18.2291
Establishment of a human hepatoma multidrug resistant cell line in vitro
Yuan Zhou, Xian-Long Ling, Shi-Wei Li, Xin-Qiang Li, Bin Yan
Yuan Zhou, Xian-Long Ling, Shi-Wei Li, Xin-Qiang Li, Bin Yan, Department of Gastroenterology, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China
Author contributions: Ling XL designed the research; Zhou Y, Li SW, Li XQ and Yan B performed the majority of experiments; Zhou Y and Ling XL analyzed the data and edited the manuscript.
Supported by The National Natural Science Foundation of China, No. 30470865; and 1520 Project of Xinqiao Hospital
Correspondence to: Xian-Long Ling, Professor, Department of Gastroenterology, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China. lingxlong@yahoo.com.cn
Telephone: +86-23-68774204 Fax: +86-23-68755604
Received: September 28, 2009
Revised: January 14, 2010
Accepted: January 21, 2010
Published online: May 14, 2010
Abstract

AIM: To establish a multidrug-resistant hepatoma cell line (SK-Hep-1), and to investigate its biological characteristics.

METHODS: 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 (IC50) values and the resistance indexes [(IC50 SK-Hep-1/CDDP)/(IC50 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.

RESULTS: The SK-Hep-1/CDDP cells (IC50 = 70.61 ± 1.06 μg/mL) was 13.76 times more resistant to CDDP than the SK-Hep-1 cells (IC50 = 5.13 ± 0.09 μg/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.

CONCLUSION: 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.

Keywords: Hepatoma; Cell line; Multidrug resistance; In vitro; Cisplatin