Related Experiment Video
Updated: Jun 16, 2026
Enrichment for Chemoresistant Ovarian Cancer Stem Cells from Human Cell Lines
Published on: September 10, 2014
Establishment of a cisplatin-induced multidrug resistance cell line SK-Hep1/DDP
1Department of Gastroenterology, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, PR China.
Background And Objective:
Multidrug resistance (MDR) is a major obstacle in the chemotherapy of cancer patients. The aim of this study was to establish a mutidrug-resistant cell line SK-Hep1/DDP and explore its molecular mechanism of the MDR.
Methods:
SK-Hep1/DDP cell line was induced by pulse treatment using a high concentration of cisplatin (DDP) in vitro. The chemoresistance indexes of cells were evaluated by CCK-8 assays. The protein of MDR1 (ABCB1), MRP1 (ABCC1), MRP2 (ABCC2) and Bax were detected by Western blotting, and the effect of MDR1 inhibitor cyclosporine A (CsA) on expression of MDR1 proteins in SK-Hep1 and SK-Hep1/DDP cell lines. Flow cytometry was performed to determine the distribution of the cell cycle and cell apoptosis ratio.
Results:
The SK-Hep1/DDP cells were 13.76 times more resistant to DDP in comparison with SK-Hep1 cells, and SK-Hep1/DDP cells also exhibited cross-resistance to many other chemotherapeutic agents (adramycin and 5-fuorouracil). MDR1, MRP1, and MRP2 protein expressions were significantly higher in the SK-Hep1/DDP than in the SK-Hep1 (P < 0. 01), but Bax was lower in the SK-Hep1/DDP than in the SK-Hep1(P < 0. 01). There was no obvious influence between SK-Hep1 and SK-Hep1/DDP cells in the expression of MDR1 by MDR1 inhibtor CsA (P > 0. 05). The percentages of cells in G(2)/M and S phase were significantly increased in SK-Hep1/DDP in comparison with those in SK-Hep1 [(20.67 +/- 5.69)% vs. (12.14 +/- 3.36)%; (42.20 +/- 2.65)% vs. (27.91 +/- 2.16)%; P < 0. 01]. After the cells were exposed to 10 μg/mL DDP for 24 h, the cell apoptosis rate of SK-Hep1/DDP was decreased in comparison with SK-Hep1, but it was increased in those with pretreatment of MDR1 inhibitor CsA as compared with those without pretreatment.
Conclusions:
A reliable multi-drug resistant human hepatoma cell line SK-Hep1/DDP is successfully established. The MDR mechanisms of this cell lines are closely related to the over-expression of MDR1 MRP1 and MRP2, lower expression of Bax and the attenuated cell apoptosis induced by chemotherapeutic agents.
Insights
This study established a multidrug-resistant (MDR) hepatoma cell line, SK-Hep1/DDP, revealing that MDR is linked to increased MDR1, MRP1, MRP2 expression, and reduced apoptosis. This provides insights into overcoming chemotherapy resistance.
Area of Science:
- Cancer Research
- Molecular Biology
- Pharmacology
Background:
- Multidrug resistance (MDR) poses a significant challenge in cancer chemotherapy.
- Developing reliable MDR cell lines is crucial for understanding resistance mechanisms and testing new therapies.
Purpose of the Study:
- To establish a multidrug-resistant human hepatoma cell line, SK-Hep1/DDP.
- To investigate the molecular mechanisms underlying the multidrug resistance in this newly established cell line.
Main Methods:
- SK-Hep1/DDP cells were generated using pulse treatment with cisplatin (DDP).
- Chemoresistance was assessed via CCK-8 assays.
- Protein expression of MDR1, MRP1, MRP2, and Bax was analyzed by Western blotting; cell cycle and apoptosis were evaluated using flow cytometry.
Main Results:
- SK-Hep1/DDP cells demonstrated a 13.76-fold resistance to DDP and cross-resistance to other agents.
- Significantly higher expression of MDR1, MRP1, and MRP2, and lower expression of Bax were observed in SK-Hep1/DDP cells compared to SK-Hep1 cells.
- SK-Hep1/DDP cells showed increased G2/M and S phase populations and reduced apoptosis rates upon DDP exposure, which was partially reversed by an MDR1 inhibitor.
Conclusions:
- A stable multidrug-resistant hepatoma cell line, SK-Hep1/DDP, was successfully established.
- The observed MDR is associated with the overexpression of MDR1, MRP1, and MRP2, decreased Bax expression, and attenuated apoptosis.
