摘要
Objective The aim of the current study was to establish an oxaliplatin-resistant hepatoma cell line(HepG2/OXA) and investigate the potential mechanisms of its drug resistance.Methods The hepatoma cell subline, HepG2/OXA, resistant to oxaliplatin(OXA), was established from a parent cell line HepG2, by stepwise exposure to gradually increasing concentrations of OXA over a half-year period. Chemosenstivity of the cytotoxic drugs, OXA, cisplatin(CDDP), adriamycin(ADM), and 5-fuorouracil(5-FU), was determined in HepG2 and HepG2/OXA cells, by the Cell counting kit-8(CCK8) assay. Cell cycle distribution of HepG2 and HepG2/OXA cells was analyzed by Flow cytometry(FCM). The expression levels of several drug resistance-related proteins, such as P-glycoprotein(P-gp), multidrug resistant protein 1(MRP1), and excision repair-cross complementing 1(ERCC1) protein in the two cell lines were tested by the western blot assay.Results The IC50 of OXA in HepG2/OXA and HepG2 were 136.84 μmol/L and 23.86 μmol/L, respectively. The resistance index(RI) was 5.34. HepG2 was also demonstrated to be cross-resistant to other antitumor agents, such as 5-FU, ADM, and CDDP. The percentage of HepG2/OXA cells in the S phase was significantly decreased compared to HepG2 cells(25.58% ± 2.36% vs 14.37% ± 2.54%, P < 0.05), while the percentage of cells in the G0/G1 and G2/M phases showed no statistical difference(respectively 55.29% ± 4.98% vs 56.73% ± 4.56%, P > 0.05, and 24.63% ± 4.81% vs 28.26% ± 3.82%, P > 0.05). The ERCC1 was found to be over expressed in HepG2/OXA cells, while there was no difference in the expressions of P-gp and MRP1 between the multiple drug resistance(MDR) phenotype cell line and its parental cell line.Conclusion HepG2/OXA showed an MDR ability; the over expression of ERCC1 might be associated with the platinum resistance of the cells, but P-gp and MRP1 are not.
Objective The aim of the current study was to establish an oxaliplatin-resistant hepatoma cell line（HepG2/OXA） and investigate the potential mechanisms of its drug resistance.Methods The hepatoma cell subline, HepG2/OXA, resistant to oxaliplatin（OXA）, was established from a parent cell line HepG2, by stepwise exposure to gradually increasing concentrations of OXA over a half-year period. Chemosenstivity of the cytotoxic drugs, OXA, cisplatin（CDDP）, adriamycin（ADM）, and 5-fuorouracil（5-FU）, was determined in HepG2 and HepG2/OXA cells, by the Cell counting kit-8（CCK8） assay. Cell cycle distribution of HepG2 and HepG2/OXA cells was analyzed by Flow cytometry（FCM）. The expression levels of several drug resistance-related proteins, such as P-glycoprotein（P-gp）, multidrug resistant protein 1（MRP1）, and excision repair-cross complementing 1（ERCC1） protein in the two cell lines were tested by the western blot assay.Results The IC50 of OXA in HepG2/OXA and HepG2 were 136.84 μmol/L and 23.86 μmol/L, respectively. The resistance index（RI） was 5.34. HepG2 was also demonstrated to be cross-resistant to other antitumor agents, such as 5-FU, ADM, and CDDP. The percentage of HepG2/OXA cells in the S phase was significantly decreased compared to HepG2 cells（25.58% ± 2.36% vs 14.37% ± 2.54%, P 〈 0.05）, while the percentage of cells in the G0/G1 and G2/M phases showed no statistical difference（respectively 55.29% ± 4.98% vs 56.73% ± 4.56%, P 〉 0.05, and 24.63% ± 4.81% vs 28.26% ± 3.82%, P 〉 0.05）. The ERCC1 was found to be over expressed in HepG2/OXA cells, while there was no difference in the expressions of P-gp and MRP1 between the multiple drug resistance（MDR） phenotype cell line and its parental cell line.Conclusion HepG2/OXA showed an MDR ability; the over expression of ERCC1 might be associated with the platinum resistance of the cells, but P-gp and MRP1 are not.
基金
Supported by grants from the National Natural Sciences Foundation of China(No.81001067)
Ministry of Science and Technology International Cooperation Project(No.S2010GR0991)
Astrazeneca Special Research Foundation for Targeted Therapy of Wu Jieping Medical Foundation(No.320.6700.09068)
关键词
房间
HEPG2
排队
癌症
P-GP
OXA
蛋白质
细胞毒素
hepatocellular carcinoma （HCC）
multidrug resistance （MDR）
excision repair-crosscomplementing 1 （ERCC1）
oxaliplatin
