联合金雀异黄素和TRAIL处理对SMMC―7721肝癌干细胞样细胞凋亡的影响.docVIP

联合金雀异黄素和TRAIL处理对SMMC―7721肝癌干细胞样细胞凋亡的影响 [摘要] 目的 肿瘤坏死因子相关凋亡诱导配体（TRAIL）选择性诱导肿瘤细胞凋亡，但是其诱导促生存反应导致耐药。金雀异黄素抑制核因子-κB（NF-κB）活性。本文旨在探讨金雀异黄素和TRAIL以及两者合用诱导人肝细胞癌SMMC-7721细胞系肝癌干细胞样细胞凋亡的作用。 方法 干细胞条件培养基超低粘附板培养SMMC-7721细胞获得第3代肿瘤球形成细胞，作为肝癌干细胞样细胞。金雀异黄素（10 μM）、TRAIL（10 ng/mL）及两者联合处理肝癌干细胞样细胞72 h；免疫酶联吸附试验（ELISA）测定细胞组蛋白/DNA碎片；碘化丙啶（PI）染色流式细胞术分析细胞凋亡率；Western blot检测细胞NF-κB（p65）蛋白表达水平。 结果 10 μM 金雀异黄素诱导肝癌干细胞样细胞组蛋白/DNA碎片水平（100% vs 152%）和凋亡率[（2.71±0.64）% vs （5.14±0.76）%]适度增高（P 0.05）；10 ng/mL TRAIL对肝癌干细胞样细胞组蛋白/DNA碎片水平（100% vs 110%）和凋亡率[（2.71±0.64）% vs （3.15±0.72）%]无明显影响；10 μM 金雀异黄素预孵育1 h，联合10 ng/mL TRAIL处理导致组蛋白/DNA碎片水平（100% vs 568%）和凋亡率[（2.71±0.64）% vs （17.3±1.27）%]显著增高（P 0.05）。10 ng/mL TRAIL诱导肝癌干细胞样细胞NF-κB（p65）蛋白表达，10 μM 金雀异黄素抑制NF-κB（p65）蛋白表达，联合金雀异黄素与TRAIL有效降低NF-κB（p65）蛋白表达水平。 结论 金雀异黄素与TRAIL协同诱导肝癌干细胞样细胞凋亡涉及NF-κB抑制。 [关键词] 肝细胞癌；肿瘤干细胞；细胞凋亡；NF-κB [中图分类号] R735.7 [文献标识码] A [文章编号] 1673-9701（2015）13-0023-03 [Abstract] Objective Tumor necrosis factor related apoptosis-inducing ligand （TRAIL） has been demonstrated that selectively induced cancer cells， but facilitated prosurvival response leading to resistance. It has been reported that genistein could inhibit nuclear factor-κB（NF-κB） exprotein and activaty. The purpose of this study was to examine the effects of genistein or TRAIL， or both on apoptosis of liver cancer stem-like cells（LCSLCs） derived from SMMC-7721 cell line. Methods The third passage sphere-forming cells（SFCs） derived from SMMC-7721 cell line which was also called LCSLCs were obtained though suspended culture in ultra-low adhesion plate with stem cell-conditioned medium. LCSLCs were treated with Genistein（10 μM） and TRAIL（10 ng/mL） alone or in combination for 72 h. Histone/DNA fragment was tested using enzyme-linked immunosorbent assay（ELISA）. And apoptosis rate was detected by flow cytometry （FCM） with propidium iodide（PI） staining； the protein expression of NF-κB（p65） was analyzed by western blot. Results The level of histone/DNA fragment（100% vs 152.6%） and apoptosis rate[（2.71±0.64）%