吉非替尼耐药的人肺腺癌HCC-827/GR细胞乙醛脱氢酶亚型表达分析

Expression of Acetaldehyde Dehydrogenase in Gefitinib-resistant Human Lung Adenocarcinoma HCC-827/GR Cells

背景与目的肿瘤的复发和耐药是肿瘤患者死亡的主要原因。乙醛脱氢酶(acetaldehyde dehydrogenase,ALDH)家族与肿瘤细胞的增殖、迁移、侵袭和耐药密切相关,且ALDH不同亚型基因在不同肿瘤细胞中有差异性表达。本实验旨在分析对吉非替尼耐药的人肺腺癌细胞HCC-827/GR ALDH亚型的表达。方法利用人肺腺癌细胞系HCC-827制备吉非替尼耐药细胞株HCC-827/GR;利用流式检测HCC-827及HCC-827/GR中ALDH的表达;采用MTT法检测ALDH抑制剂二乙氨基苯甲醛(diethyllaminaldehyde,DEAB)处理前后HCC-827/GR细胞的增殖能力和对吉非替尼的敏感性;利用q RT-PCR检测HCC-827与HCC-827/GR细胞中ALDH各亚型在m RNA水平的表达。结果与HCC-827细胞相比,ALDH在吉非替尼耐药的细胞株HCC-827/GR的阳性率增加;经100μmol/L DEAB处理后,HCC-827/GR细胞增殖能力下降;与HCC-827细胞相比,ALDH1A1和ALDH1L1在HCC-827/GR细胞中m RNA表达水平增高;ALDH3B2表达降低。结论 ALDH具有检测吉非替尼耐药的人肺腺癌细胞的标志分子的潜力,其中ALDH1A1可能参与人肺腺癌细胞对吉非替尼耐药性的形成过程。

Background and objective Tumor recurrence and drug resistance are the main causes of death in tumor patients. The family of acetaldehyde dehydrogenase（ALDH） is closely related to the proliferation, migration, invasion and resistance of tumor cells, and different ALDH subtypes are expressed in different tumor cells. The aim of this study is to elucidate the ALDH subtype in human lung adenocarcinoma HCC-827/GR cells, which resistant to the gefitinib. Methods The human lung adenocarcinoma HCC-827 cells were used to generate the gefitinib-resistant HCC-827/GR cells; the expression of ALDH subtype in either HCC-827 or HCC-827/GR was detected by flow cytometry; The proliferative capacity and sensitivity to gefitinib of hcc-827/GR cells were analyzed by MTT assay before and after treatment with 100 μmol/L diethyllaminaldehyde（DEAB）; Real-time quantitative PCR was used to detect the expression of ALDH subtypes at mRNA levels in hcc-827 cells and hcc-827/GR cells. Results Compared with HCC-827 cells, the positive rate of ALDH in HCC-827/GR cells increased. The proliferation ability of HCC-827/GR cells decreased after treatment with 100 μmol/L DEAB. Compared with HCC-827 cells, the expression of ALDH1A1 and ALDH1L1 mRNA wasincreased in hcc-827/GR cells, but the ALDH3B2 expression was decreased. Conclusion ALDH might be used as a molecular biomarker to test the gefitinib-resistant to lung adenocarcinoma cancer cells, and the ALDH1A1 may play a role in gefitinib resistance in lung cancer.