| Ping Qiu,Gang Chen,Yuhong Dai,Hong Qiu. Establishment and characterization of an oxaliplatin-resistant hepatic cancer cell line. Oncol Transl Med, 2018, 4: 48-53. |
| 肝癌耐OXA细胞系的建立及特性鉴定 |
| Establishment and characterization of an oxaliplatin-resistant hepatic cancer cell line |
| Received:April 06, 2018 Revised:May 17, 2018 |
| DOI：10.1007/s10330-018-0267-7 |
| 中文关键词: 肝细胞癌；多药耐药；ERCC1；奥沙利铂 |
| 英文关键词: hepatocellular carcinoma (HCC); multidrug resistance (MDR); excision repair-cross complementing 1 (ERCC1); oxaliplatin |
| 基金项目: |
| Author Name | Affiliation | E-mail | | Ping Qiu* | Oncology Department,Jingzhou Central Hospital | tjqiuhong@163.com | | Gang Chen | Integration Traditional Chinese Medicine and Weste,Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology | | | Yuhong Dai | Department of Oncology,Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology | | | Hong Qiu | Department of Oncology，Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology | |
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| 中文摘要: |
| 目的: 国际多中心III期临床试验EACH研究证实肝细胞癌可以从FOLFOX4化疗方案中获益。然而，肝癌细胞的获得性耐药是治疗失败的原因之一。寻找肝癌细胞对奥沙利铂获得性耐药的内在机制有助于提高肝癌化疗获益。
方法：肝癌奥沙利铂耐药细胞系HepG2/OXA由亲代细胞OXA暴露在浓度逐步升高的OXA共培养诱导获得。CCK8检测分析该细胞系对细胞毒类药物OXA、DDP、ADM及5-FU的化疗敏感性。流式细胞学检测亲代细胞及耐药细胞系的细胞周期分布差异。Western blot方法检测部分耐药蛋白P-gp、 MRP1及ERCC1在两种细胞系中的表达差异。
结果：HepG2/OXA 及 HepG2对OXA的IC50分别为136.84 umol/L 及23.86 μmol/L。相对耐药RI为5.34。HepG2/OXA对5-FU、OXA、CDDP也产生了交叉耐药。与HepG2相比，The cell percentage in S phases of HepG2/OXA细胞系的S期细胞比例明显降低 ( (14.37% ±2.54% vs 25.58% ± 2.36%, P < 0.05) )，而G0/G1及G2/M期细胞比例没有显著性差异 (分别为 55.29% ± 4.98% vs 56.73% ± 4.56%, P > 0.05, 24.63% ± 4.81% vs 28.26% ± 3.82%, P > 0.05)。HepG2/OXA细胞的ERCC1蛋白表达明显增加，而P-gp及MRP1 表达在两种细胞系之间无差异。
结论：人肝癌细胞系HepG2/OXA 是具有多药耐药的细胞系，ERCC1的过表达与该细胞系的多药耐药具有相关性。 |
| 英文摘要: |
| 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 anti-tumor 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.
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