Expression profile of stem cell markers and ABC transporters in 5-fluorouracil resistant Hep-2 cells
Expression profile of stem cell markers and ABC transporters in 5-fluorouracil resistant Hep-2 cells
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DOI：
10.1007/s11033-020-05633-x
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发表时间：
2020-07-05
影响因子：
2.8
通讯作者：
Karatas, Omer Faruk
中科院分区：
文献类型：
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作者：
Duz, Mehmet Bugrahan;Karatas, Omer Faruk
喉鳞状细胞癌细胞对传统治疗方案的耐药仍是导致治疗失败的主要原因。在本研究中，我们的目的是研究在5-氟尿嘧啶（5-FU）耐药的喉Hep-2细胞中ATP结合盒（ABC）转运蛋白和干细胞标志物的表达谱。我们用逐步增加剂量的5-FU处理亲本Hep-2细胞近1年，以开发对不同水平的5-FU浓度具有抗性的5-FU抗性亚系（4个亚系对1、2、4和8倍的5-FU具有抗性）。然后，我们使用定量逆转录聚合酶链反应（qRT-PCR）检测ABC转运蛋白家族的10个基因和4个干细胞相关标志物的表达水平，以找出这些标志物与化疗耐药性之间的潜在关系。我们发现，从5-FU耐药获得开始，干细胞相关标志物的表达升高。当亲本Hep-2细胞对较高剂量的5-FU产生耐药性时，它们的表达逐步升高。在5-FU耐药的Hep-2细胞中，测试的ABC转运蛋白（ABCA 5、ABCB 1、ABCB 6、ABCC 1、ABCC 2、ABCC 3、ABCC 5、ABCC 10和ABCF 2以及ABCG 2）的表达也失调。虽然他们的表达保持不变，在开始收购的阻力，表达的ABC转运蛋白，除了从ABCB 6显着增加时，细胞变得耐高剂量的5-FU。我们的研究结果表明，富集的干细胞特性和上调ABC转运蛋白可能是喉癌细胞化疗耐药的关键因素之一。
Resistance of laryngeal squamous cell carcinoma cells to traditional therapeutic regimens still remains to be a major reason for therapeutic failure in patients. In this study, we aimed at investigating the expression profiles of ATP-binding cassette (ABC) transporters and stem cell markers in 5-fluorouracil (5-FU) resistant laryngeal Hep-2 cells. We treated parental Hep-2 cells, with stepwise increased doses of 5-FU for almost 1 year to develop 5-FU resistant sub-lines with resistance against varying levels of 5-FU concentrations (4 sub-lines resistant to 1, 2, 4, and eightfold of 5-FU). Then, we measured the expression levels of 10 genes from ABC transporters family and 4 stem cell associated markers using quantitative reverse transcription polymerase chain reaction (qRT-PCR) to find out a potential relationship between these markers and chemoresistance. We found that stemness-associated markers had elevated expressions from the beginning of 5-FU resistance acquisition. Their expressions elevated stepwise while parental Hep-2 cells got resistance to higher doses of 5-FU. Expressions of tested ABC transporters (ABCA5, ABCB1, ABCB6, ABCC1, ABCC2, ABCC3, ABCC5, ABCC10 and ABCF2, and ABCG2) were also deregulated in 5-FU resistant Hep-2 cells. Although their expressions remained unaltered at the beginning of acquisition of resistance, expressions of ABC transporters except from ABCB6 increased significantly when cells became resistant to higher doses of 5-FU. Our results suggest that enrichment of cells with stemness characteristics and upregulation of ABC transporters might be amongst the crucial contributors of chemoresistance in laryngeal cancer cells.