The effects of multidrug resistance on cell proliferation, apoptosis and invasion activity and the expression of mitogen-activated protein kinase in human hepatocellular cancer cells
10.3781/j.issn.1000-7431.2012.07.005
- Author:
Xin YAN
1
Author Information
1. Department of Clinical Medicine
- Publication Type:Journal Article
- Keywords:
Liver neoplasms;
Mitogen-activated protein kinases;
Multidrug resistance-associated proteins
- From:
Tumor
2012;32(7):507-515
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To establish multiple human hepatocellular carcinoma (HCC) multidrug-resistant cell lines induced by chemotherapeutic drugs, and to investigate the effects of multidrug-resistance on cell proliferation, apoptosis and invasion activity and the relationship between the multidrug-resistance and the expression of mitogen-activated protein kinase (MAPK) pathway. Methods: Three different human HCC cell strains HepG2, SMMC-7721 and BEL-7402 were established by using pulse treatment with high concentration of adriamycin (ADM) or treatment with ADM of low concentration gradually increased. The difference between the resistant cells and the parental cells was evaluated. The drug sensitivity was tested by ATP bioluminescence to calculate the resistance index (RI). The expressions of multidrug resistance-related genes including P-glycoprotein (P -gp ), multidrug resistance-associated protein 1 (MRP 1), lung cancer resistance protein 1 (LRP 1), breast cancer resistance protein (BCRP ), glutathione-S - transferase-π (GST -π), topoisomerase IIβ (ToPo IIβ) and protein kinase C (PKC ) and their proteins were detected by RT-PCR and immunohistochemistry (IHC), respectively. The expressions of cell proliferative activity-related Ki-67 and proliferating cell nuclear antigen (PCNA) were detected by IHC. The cell cycle and apoptosis rate were detected by flow cytometry (FCM), and the change of cell invasion ability was detected by Transwell assay. The expressions of ERK 1, ERK 2, ERK 5, JNK 1, JNK 2 and p 38a genes and their proteins in MAPK pathway were detected by RT-PCR and Western boltting, respectively. Results: Six ADMresistant cell strains were established. Compared to parental cells, the RIs of all resistant cells were over 5 with increased resistance to multiple chemotherapeutic drugs. The expressions of MDR genes and their proteins were increased 2-10 times, especially for the cells induced by pulse treatment with high concentration of ADM. The expressions of Ki-67 and PCNA proteins were increased over 20 times in the resistant cells. The cell cycles were arrested at phase S, and the invasive activities in vitro were increased 1.3-2.5 times. The apoptosis rates were decreased by over 60% after ADM intervention. The expressions of MAPK pathway-related genes and proteins were increased in varying degrees, especially for ERK1 and ERK2. Conclusion: ADM can induce the multidrug-resistance in multiple HCC cell strains and increase the cell proliferation activity, alter the cell cycle distribution, strengthen the anti-apoptosis effect and enhance the invasive ability of the HCC cells. The multidrug-resistance maybe associated with the upregulation of MAPK pathway. Copyright © 2012 by TUMOR.
