摘要
目的研究核因子-κB(NF-κB)抑制剂吡咯烷二硫代氨基甲酸盐(PDTC)联合TNF-α对宫颈癌HeLa细胞生长及凋亡的影响,并初步探讨其作用机制。方法应用噻唑蓝(MTT)法检测不同浓度PDTC、TNF-α及两者联合作用对HeLa细胞生长抑制率的影响;采用光镜及DAPI法观察细胞形态学变化及凋亡情况;Westernblot法检测HeLa细胞A20和caspase-3蛋白的表达。结果 PDTC(50,100μmol/L)或TNF-α(30,60,120mg/L)可明显抑制细胞的增殖;低浓度PDTC(25μmol/L)不影响细胞增殖,但与TNF-α联合应用与单用TNF-α比较,细胞增殖抑制率明显增加(P<0.01);光镜与DAPI染色结果显示,不同浓度药物组的细胞均有明显的细胞凋亡特征,且联合用药组细胞凋亡率增高(P<0.01);PDTC(25μmol/L)与TNF-α(60mg/L)联合用药与单用TNF-α(60mg/L)或PDTC(50μmol/L)比较,胞质A20蛋白表达无明显变化,但caspase-3蛋白的表达明显增加(P<0.05)。结论 PDTC可增强TNF-α对宫颈癌HeLa细胞的毒性作用,其机制可能与PDTC抑制TNF-α诱导的核转录因子NF-κB的活性,最终上调凋亡蛋白caspase-3表达有关。
Objective To detect the effect of PDTC(the selective inhibitor of NF-κB) combined with TNF-α on the growth and apoptosis of human cervical cancer HeLa cells,and the related mechanisms.Methods After treated by different doses of PDTC,TNF-α or combined with the two on human cervical cancer HeLa cells,MTT assay was used to measure the growth inhibition of HeLa cells.Light microscopy and DAPI method were used to observe the morphological changes and apoptosis.The protein expression of A20 and caspase-3 were detected by Western blot assay.Results PDTC(50,100 μmol/L) or TNF-α(30,60,120 mg/L) significantly inhibited the growth of HeLa cells.Low dose of PDTC(25 μmol/L) did not affect cell growth,however,its combination with TNF-α significantly increased the cell growth inhibition rate(P<0.01) as compared with TNF-α.Cells appeared apoptotic morphological changes and the combination group increased the percentage of cell apoptosis(P<0.01).In addition,TNF-α(60 mg/L) in combination with PDTC(25 μmol/L) group had no significant change in cytoplasmic protein A20,but increased the expression of caspase-3 protein(P<0.05),compared with either TNF-α(60 mg/L)or PDTC(50 μmol/L) group.Co nclusions PDTC evidently enhances the cytotoxicity of TNF-α on cervical cancer cell line HeLa cells,which may be performed by inhibiting the activity of the TNF-α-induced nuclear transcription factor NF-κB and ultimately up-regulating of caspase-3 expression.
出处
《中华临床医师杂志（电子版）》
CAS
2011年第12期3496-3502,共7页
Chinese Journal of Clinicians(Electronic Edition)
