TRAIL联合PDTC对SGC-7901细胞生长抑制及凋亡诱导实验探究.doc

TRAIL联合PDTC对SGC-7901细胞生长抑制及凋亡诱导实验探究 　【摘要】 目的 观察肿瘤坏死因子相关凋亡诱导配体（TRAIL）及TRAIL联合核转录因子核因子-κB（NF-κB）活性特异性抑制剂吡咯烷二硫代氨基甲酸盐(PDTC)对胃癌细胞SGC-7901的生长抑制和凋亡诱导作用，并寻找逆转TRAIL耐药的方法。方法 采用不同浓度的TRAIL及TRAIL联合PDTC作用于胃癌细胞后，MTT法检测细胞生长抑制率，流式细胞仪检测细胞的凋亡率，免疫细胞化学染色方法观察药物作用前后NF-κB表达情况。结果 单独使用TRAIL时，随着浓度从50 μg/L增加到500 μg/L，细胞的生长抑制率和凋亡率逐渐增加（Plt;0.05）,但这种作用是非线性的。TRAIL联合0.05 μmol/L、0.1 μmol/L PDTC后肿瘤细胞的生长抑制率和凋亡率较单用TRAIL显著下降（Plt;0.05）；而联合1 μmol/L、10 μmol/L PDTC后其作用较上有所上升（Plt;0.05）。联合作用组 p65蛋白在胞核中染色的强度低于单纯使用TRAIL组（Plt;0.05）。结论 TRAIL对胃癌细胞株SGC-7901具有一定程度的生长抑制和凋亡诱导作用；NF-κB信号转导途径可能具有双向调节作用，大剂量PDTC可能逆转胃癌细胞对TRAIL的耐药。 【关键词】 肿瘤坏死因子相关凋亡诱导配体；吡咯烷二硫代氨基甲酸盐；胃癌细胞；凋亡；核因子-κB Abstract：Objective To observe the effect of a combination of pyrrolidine dithiocarbonate (PDTC), a specific inhibitor of NF-κB and tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) on the apoptosis of gastric cancer cells and their effect on the expression of NF-κB, and to search for the approaches to reversing resistance to TRAIL in human stomach cancer cells. Methods Various concentrations of TRAIL and PDTC were added in exponential growth SGC-7901 gastric cancer cells. MTT assays were used to measure the growth inhibitory effect of combined use of TRAIL and PDTC or alone. Apoptosis of gastric cancer cell SGC-7901 was analyzed with PI staining method and flow cytometry (FCM). The expression of NF-κB was observed using immunohistochemical staining method. Results After the SGC-7901 gastric cancer cells were exposed to TRAIL at a dose of 50 μg/L to 500 μg/L, the growth inhibitory rate and apoptosis rate increased (Plt; 0.05), but there was no linearity for this dose-effect relationship. Following TRAIL treatment in combination with 0.05 μmol/L and 0.1 μmol/L PDTC, the growth inhibitory rate of and apoptosis rate significantly decreased compared with treatment with TRAIL or PDTC alone (Plt;0.05), while following TRAIL treatment in combination with PDTC 1 μmol/L and 10 μmol/L, the growth inhibitory rate and apoptosis rates increas