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Development and characterization of multidrug resistant human hepatocarcinoma cell line in nude mice
Bao-Jin Zhai1, Ze-Yong Shao, Chun-Liang Zhao
1Clinical Center for Tumor Therapy of 2nd Affiliated Hospital, Box 153, Institute of Ultra-sonic Engineering in Medicine, Chongqing University of Medical Sciences, 1 Medical College Road, Chongqing 400016, China.
Aim:
To establish a multidrug resistant (MDR) cell sub-line from the human hepatocarcinoma cell line (HepG2) in nude mice.
Methods:
HepG2 cell cultures were incubated with increasing concentrations of adriamycin (ADM) to develop an ADM-resistant cell subline (HepG2/ADM) with cross-resistance to other chemotherapeutic agents. Twenty male athymic BALB/c-nu/nu mice were randomized into HepG2/nude and HepG2/ADM/nude groups (10 in each group). A cell suspension (either HepG2 or HepG2/ADM) was injected subcutaneously into mice in each group. Tumor growth was recorded, and animals were sacrificed 4-5 wk after cell implantation. Tumors were prepared for histology, and viable tumor was dispersed into a single-cell suspension. The IC50 values for a number of chemotherapeutic agents were determined by 2, 3-bis (2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide inner salt (MTT) assay. Rhodamine-123 retention/efflux and the level of resistance-associated proteins were determined by flow cytometry. The mRNA expression of mdr1, mrp and lrp genes was detected using reverse transcriptase polymerase chain reaction (RT-PCR) in HepG2/nude and HepG2/ADM/nude groups.
Results:
The appearances of HepG2/nude cells were slightly different from those of HepG2/ADM/nude cells. Similar tumor growth curves were determined in both groups. A cross-resistance to ADM, vincristine, cisplatin and 5-fluorouracil was seen in HepG2/ADM/nude group. The levels of P-glycoprotein and multidrug resistance-associated proteins were significantly increased. The mRNA expression levels of mdr1, mrp and lrp were higher in HepG2/ADM/nude cells.
Conclusion:
ADM-resistant HepG2 subline in nude mice has a cross resistance to chemotherapeutic drugs. It may be used as an in vivo model to investigate the mechanisms of MDR, and explore the targeted approaches to overcoming MDR.
Insights
Researchers developed an adriamycin-resistant HepG2 cell line in nude mice, demonstrating cross-resistance to multiple chemotherapeutic drugs. This new multidrug-resistant (MDR) model aids in studying MDR mechanisms and developing targeted therapies.
Area of Science:
- Oncology
- Pharmacology
- Cell Biology
Background:
- Hepatocellular carcinoma (HCC) poses a significant health challenge.
- Multidrug resistance (MDR) in cancer limits the efficacy of chemotherapy.
- Developing reliable in vivo models is crucial for studying MDR.
Purpose of the Study:
- To establish a multidrug-resistant (MDR) cell sub-line from the human hepatocarcinoma cell line (HepG2) in nude mice.
- To characterize the cross-resistance profile of the developed MDR cell line.
- To validate its utility as an in vivo model for MDR research.
Main Methods:
- HepG2 cells were incubated with increasing adriamycin (ADM) concentrations to create an ADM-resistant subline (HepG2/ADM).
- HepG2/ADM and parental HepG2 cells were implanted into nude mice to establish tumor xenografts.
- Chemosensitivity (IC50), drug efflux proteins (flow cytometry), and gene expression (RT-PCR) were analyzed.
Main Results:
- The HepG2/ADM subline exhibited cross-resistance to ADM, vincristine, cisplatin, and 5-fluorouracil.
- Increased levels of P-glycoprotein and multidrug resistance-associated proteins were observed.
- Elevated mRNA expression of mdr1, mrp, and lrp genes was detected in the resistant cells.
Conclusions:
- The established adriamycin-resistant HepG2 subline in nude mice effectively models cross-resistance to chemotherapeutic drugs.
- This in vivo model is suitable for investigating the mechanisms underlying MDR.
- It provides a platform for exploring novel targeted strategies to overcome MDR in cancer treatment.
