【摘要】 背景:丁酸钠是一种组蛋白去乙酰化酶抑制剂,能够抑制多种肿瘤细胞的增殖、诱导细胞凋亡及分化,但其联合肿瘤坏死因子相关凋亡诱导配体对肺癌干细胞的影响尚未见报道。目的:研究丁酸钠单独或联合给药对肺癌干细胞生物学行为的影响。方法:利用免疫磁珠细胞分选技术从人肺腺癌A549细胞中分选CD133~+肺癌干细胞,将CD133~+肺癌干细胞分4组培养,对照组以DMEM/F12培养基培养,丁酸钠组以含5 mmol/L丁酸钠的DMEM/F12培养基培养,TRAIL组以含50μg/L肿瘤坏死因子相关凋亡诱导配体的DMEM/F12培养基培养,联合组以含5 mmol/L丁酸钠+50μg/L肿瘤坏死因子相关凋亡诱导配体的DMEM/F12培养基培养。检测培养96 h内的细胞增殖、培养24 h后的细胞凋亡、培养48 h内的细胞迁移能力,以及培养48 h后的多能性转录因子Oct4、Sox2及Nanog蛋白表达。结果与结论:(1)细胞增殖:联合组培养不同时间点的细胞增殖抑制率显著高于丁酸钠组、TRAIL组(P<0.05);(2)细胞凋亡:丁酸钠组、TRAIL组、联合组细胞凋亡率高于对照组(P<0.05),联合组高于丁酸钠组、TRAIL组(P<0.05);(3)细胞迁移能力:丁酸钠组、TRAIL组、联合组细胞划痕距离大于对照组(P<0.05),联合组大于丁酸钠组、TRAIL组(P<0.05);(4)多能性转录因子表达:丁酸钠组、TRAIL组、联合组多能性转录因子Oct4、Sox2及Nanog蛋白表低于对照组(P<0.05),联合组低于丁酸钠组、TRAIL组(P<0.05);(5)结果表明:丁酸钠与肿瘤坏死因子相关凋亡诱导配体联合用药,对肺癌干细胞有协同抑制作用。更多还原

【Abstract】 BACKGROUND: Sodium butyrate, a histone deacetylase inhibitor, can inhibit cell proliferation, and induce apoptosis and differentiation of various cancer cells. However, the role of sodium butyrate combined with tumor necrosis factor-related apoptosis-inducing ligand(TRAIL) on lung cancer stem cells remains unclear. OBJECTIVE: To explore the effect of sodium butyrate combined with TRAIL on biological behaviors of lung cancer stem cells. METHODS: Magnetic bead separation was used to separate lung cancer stem cells(CD133~+) from human lung adenocarcinoma A549 cells. After the lung cancer stem cells were treated with simple DMEM/F12, DMEM/F12 containing sodium butyrate(5 mmol/L), TRAIL(50 μg/L) or sodium butyrate combined with TRAIL, the cell proliferation within 96 hours of culture was determined by MTT assay; the apoptosis within 24 hours of culture was measured by flow cytometry; the cell migration within 48 hours of culture was detected by cell scratch test; the expression levels of pluripotent transcription factors(Oct4, Sox2 and Nanog) within 48 hours of culture were detected using western blot analysis. RESULTS AND CONCLUSION: The CD133~+ lung cancer stem cells were successfully enriched from human lung adenocarcinoma A549 cells. MTT assay showed that sodium butyrate and TRAIL significantly inhibited the proliferation of lung cancer stem cells(P < 0.05), and the combination effect was even stronger(P < 0.05). Results from flow cytometry analysis and scratch test showed that sodium butyrate or TRAIL induced apoptosis and inhibited cell migration of lung cancer stem cells(P < 0.05), and the combination of sodium butyrate and TRAIL showed a stronger effect(P < 0.05). In addition, the expression levels of Oct4, Sox2 and Nanog were significantly down-regulated by sodium butyrate(P < 0.05), TRAIL or sodium butyrate combined with TRAIL, and the combination effect was stronger(P < 0.05). In conclusion, sodium butyrate and TRAIL have synergistic effects on lung cancer stem cells, indicating a new way for treatment of lung cancer.更多还原