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目的研究Af116609基因转染胃癌耐药细胞系SGC7901/VCR后,对胃癌细胞耐药性的影响。方法采用RT-PCR法克隆Af116609基因,用Northernblot检测其差异表达,以基因转染技术调节其表达,通过体外药敏实验观察其对胃癌耐药表型的影响。结果从胃癌耐药细胞SGC7901/VCR中克隆到Af116609基因的CDS序列327bp,该序列与GenBank登录的一致。Northernblot结果显示,该基因在耐药细胞高表达。体外药敏实验发现,转染正义真核表达载体的药敏细胞中该基因上调,对长春新碱、5-氟脲嘧啶和阿糖胞苷的耐药性增加,而对阿霉素的抗性有损害,对氨甲喋呤的抗性无明显影响;转染反义真核表达载体的耐药细胞中该基因下调,对上述5种药物的抗性均减弱。结论Af116609基因与胃癌MDR表型相关,可作为逆转胃癌MDR的候选靶分子。
Objective To investigate the effect of Af116609 gene on the drug resistance of gastric cancer cells after transfection with gastric cancer cell line SGC7901 / VCR. Methods The gene of Af116609 was cloned by RT-PCR and the expression of Af116609 was detected by Northern blot. The gene expression was detected by gene transfection technique. The effect of Af116609 gene on the drug resistance phenotype of gastric cancer was observed by drug sensitivity test in vitro. Results The CDS sequence of Af116609 gene was cloned from SGC7901 / VCR gastric cancer cell line 327bp, which was identical to that of GenBank. Northern blot results showed that the gene was highly expressed in drug-resistant cells. In vitro susceptibility experiments showed that the gene transfection of sense eukaryotic expression vector of the drug-sensitive cells increased resistance to vincristine, 5-fluorouracil and cytarabine resistance increased, and the resistance to doxorubicin Sexual damage, no significant impact on methotrexate resistance; transfected antisense eukaryotic expression vector resistant cells in the gene down regulation of the five drugs were weakened. Conclusion The Af116609 gene is associated with MDR phenotype in gastric cancer and can be used as a potential target for reversing MDR in gastric cancer.