Study of the Characterization of Side Population Cells in Endometrial Cancer Cell Lines: Chemoresistance, Progestin Resistance, and Radioresistance
Study of the Characterization of Side Population Cells in Endometrial Cancer Cell Lines: Chemoresistance, Progestin Resistance, and Radioresistance
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DOI：
10.3389/fmed.2020.00070
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发表时间：
2020-03-18
影响因子：
3.9
通讯作者：
Li, Xiao-ping
中科院分区：
文献类型：
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作者：
Liu, Bing-jie;Xu, Qi-ying;Li, Xiao-ping
简介：放射治疗、铂类药物联合紫杉醇化疗和/或内分泌治疗是子宫内膜癌术后重要的辅助治疗手段。然而，对它们的抵抗仍然不清楚。本研究旨在从EC细胞系中分离SP细胞的特性，研究其对紫杉醇耐药、对紫杉醇耐药和对辐射耐药的机制，为EC的研究提供基础。采用免疫细胞化学方法检测HEC-1A SP细胞和非SP细胞中乳腺癌耐药蛋白（BCRP）的表达，MTS法比较SP细胞和非SP细胞的放射抗性和紫杉醇抗性。应用流式细胞术检测MPA对石川、石川SP和石川非SP细胞凋亡的影响。结果：HEC-1A、石川和RL 95 -2中SP细胞比例分别为1.44 ± 0.93%、2.86 ± 3.09%和2.87 ± 1.29%。HEC-1A中SP细胞的克隆形成效率高于非SP细胞[（6.02 +/- 1.17）vs.（0.53 +/- 0.20）%，P = 0.001]，HEC-1A中SP细胞和非SP细胞的致瘤率存在显著差异（87.5vs. 12.5%）。SP细胞的BCRP表达水平（P = 0.001）以及对紫杉醇和放射的抵抗力（P < 0.05）高于非SP细胞。MPA处理后，石川组、石川SP组和石川非SP组细胞凋亡率分别为（4.64 ± 0.18）%、（4.01 ± 0.43）%和（9.3 ± 0.67）%，差异有统计学意义（P = 0.05），且石川组Caspase-3表达高于石川SP组。结论：SP细胞在不同EC细胞系中有明显的富集，这些SP细胞对紫杉醇、甲孕酮和放射治疗有较强的抵抗力。BCRP在SP细胞中的过度表达可能是导致其对紫杉醇、紫杉醇和放射治疗抵抗的原因，其机制可能与细胞凋亡和自噬活性有关。
Introduction: Radiotherapy, combined regimens as platinum-paclitaxel chemotherapy and/or endocrine therapy is an important adjuvant treatment after surgery for endometrial cancer (EC). While, the resistance to them remain unclear. In our study, to separate the characteristics of side population (SP) cells from EC cell lines, study the mechanism of Taxol-resistance, progestin resistance and radioresistanc, and provide the basic for EC.Methods: SP cells from EC cell lines HEC-1A, Ishikawa and RL95-2 were separated by Hoechst 33342 staining and flow cytometry analysis. The expression of breast cancer resistance protein (BCRP) in SP cells and non-SP cells from HEC-1A was examined by immunocytochemistry, and the radiation-resistant and Taxol-resistant characteristics of SP cells and non-SP cells were compared by MTS. Ishikawa, Ishikawa-SP, and Ishikawa-non-SP cells incubated with MPA were selected for cell apoptosis assays by using flow cytometry. The expression of caspase-3 was examined by immunocytochemistry, and autophagy was detected by MDC staining.Results: Small proportions of SP cells, namely, 1.44 +/- 0.93%, 2.86 +/- 3.09%, and 2.87 +/- 1.29%, were detected in HEC-1A, Ishikawa and RL95-2, respectively. There was a stronger clone formation efficiency for the SP cells than for non-SP cells in HEC-1A [(6.02 +/- 1.17) vs. (0.53 +/- 0.20)%, P = 0.001], and there was a significant difference in the rate of tumourigenicity between the SP cells and non-SP cells in HEC-1A (87.5 vs. 12.5%). There were higher levels of BCRP expression (P = 0.001) and resistance to Taxol and radiation (P < 0.05) in the SP cells than in non-SP cells. After MPA treatment, the apoptosis rates were significantly different among the Ishikawa, Ishikawa-SP and Ishikawa-non-SP groups [(4.64 +/- 0.18)%, (4.01 +/- 0.43)%, and (9.3 +/- 0.67)%; (P = 0.05)], and the expression of Caspase-3 in the Ishikawa group was higher than that in Ishikawa-SP group. The autophagic activity of the Ishikawa-SP cells was the strongest, while the autophagic activity of Ishikawa-non-SP was the weakest.Conclusions: There is a significant enrichment in SP cells among different EC cell lines, and these SP cells be more resistant to Taxol, MPA and radiation therapy. The overexpression of BCRP among SP cells may be the cause of resistance to Taxol, progestin and radiotherapy, which may be related to apoptosis and autophagic activity.