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目的 探讨CD95系统在胰腺癌细胞对化疗敏感性中的作用。方法 通过脂质体介导将CD95基因转入胰腺癌细胞株SW 1990 ,G4 18筛选转导CD95基因的细胞克隆 ,Northern blot和Western blot检测转导细胞中CD95的表达 ;MTT法观察转导细胞对氟尿嘧啶 (5 FU)、阿霉素 (ADM )、双氟脱氧胞苷 (GCB)及与抗CD95单克隆抗体联合治疗的反应 ;流式细胞仪 (FCM)检测化疗药诱导转导细胞凋亡的作用。结果 CD95基因转导的SW 1990细胞可稳定地过表达CD95 ,其mRNA与蛋白的表达均较未转导及空白对照组显著增高 ,抗CD95单抗可阻滞转导细胞的生长。临床相关药物浓度的化疗药对转导细胞的生长抑制作用明显高于未转导细胞 ,加入抗CD95单抗可显著增强化疗药的作用。ADM诱导胰腺癌细胞凋亡作用较明显 ,且转导细胞显著高于未转导细胞。结论 转导CD95基因可提高SW1990细胞对化疗药的敏感性 ,部分逆转其耐药性。化疗药与抗CD95单抗联合应用对胰腺癌细胞具有协同作用
Objective To investigate the role of CD95 in chemosensitivity of pancreatic cancer cells. Methods The CD95 gene was transfected into pancreatic cancer cell line SW 1990 by lipofectamine. The cell clones transduced with CD95 were screened by G4 18. The expression of CD95 was detected by Northern blot and Western blot. The expression of CD95 was detected by MTT assay The response of 5 FU, ADM, GCB and anti-CD95 monoclonal antibody treatment was detected by flow cytometry (FCM) Role. Results The CD95 gene-transduced SW 1990 cells stably overexpressed CD95. The mRNA and protein expressions of CD95 were significantly higher than those of untransduced and blank control cells. Anti-CD95 mAb could block the growth of transduced cells. The chemotherapeutic drugs with clinically relevant drug concentrations have a significantly higher inhibitory effect on transduced cells than non-transduced cells. The addition of anti-CD95 mAb can significantly enhance the effect of chemotherapeutic drugs. ADM induced apoptosis of pancreatic cancer cells was more obvious, and the transduced cells were significantly higher than the non-transduced cells. Conclusion Transfection of CD95 gene can increase the sensitivity of SW1990 cells to chemotherapeutic drugs and partially reverse its drug resistance. Chemotherapy drugs and anti-CD95 monoclonal antibody combination has synergistic effect on pancreatic cancer cells