可溶性CD40配体对HL-60细胞增殖影响及其机制探讨  

Effect of soluble CD40 ligand on proliferation of HL-60 cells and its mechanism

作　　者：封忠昕[1] 陈琦[2] 肖建辉 冯进[1] 

机构地区：[1]遵义医学院附属医院检验科,贵州遵义563003 [2]遵义医学院附属医院血液内科,贵州遵义563003 [3]贵州省细胞工程重点实验室,贵州遵义563003

出　　处：《中华肿瘤防治杂志》2016年第2期69-73,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：贵州省社会发展攻关项目(黔科合SY字[2014]3025);贵州省科技厅联合基金(黔科合J字LKZ[2013]55)

摘　　要：目的 Survivin和IL-8在急性髓系白血病(acute myeloid leukemia,AML)等多种恶性肿瘤中高表达,与白血病的治疗效果及预后密切相关。本研究分析可溶性CD40配体(soluble CD40L,sCD40L)对人白血病HL-60细胞增殖的影响,探讨其可能作用机制。方法 2、4和6μg/mL 3组不同浓度的sCD40L作用HL-60细胞24、48和72h,采用MTT法检测sCD40L孵育HL-60细胞体外增殖的影响;通过RT-PCR检测sCD40L对HL-60细胞Survivin mRNA表达的影响;采用ELISA测定sCD40L对HL-60细胞分泌IL-8的影响。结果 sCD40L3组不同浓度(2、4和6μg/mL)均能显著抑制HL-60细胞增殖,并呈剂量依赖关系,不同浓度之间差异有统计学意义,F=150.076,P<0.05;不同时间之间差异有统计学意义,F=358.969,P<0.05。在24、48和72h,以作用48h时抑制效果最明显。RT-PCR检测结果显示,以6.00μg/mL的sCD40L作用时Survivin mRNA表达量最低,时间以作用48h时HL-60细胞中Survivin mRNA表达量最低,其表达量为89.7±6.2,与As2O3阳性对照组比较,差异有统计学意义,P<0.05。ELISA检测结果显示,以6.00μg/mL的sCD40L作用时IL-8表达量最低,时间以作用48h时HL-60细胞中IL-8表达量最低,其表达量为380.2±29.5,与As2O3阳性对照组比较,差异有统计学意义,P<0.05。结论 sCD40L在体外能够明显抑制HL-60细胞增殖,具有浓度依赖性,且与作用时间有关。sCD40L抑制HL-60细胞增殖与Survivin及IL-8表达相关,并可能与其下调有关。OBJECTIVE Survivin and IL-8were highly expressed in acute myeloid leukemia and a variety of malignant tumors,and closely related to the therapeutic effect and prognosis of leukemia.The objective of this study was investigate the effect of sCD40 Lon proliferation of HL-60 cells and expressions of IL-8and Survivin,and explore its possible mechanism.METHODS HL-60 cells were cultered with three different concentrations of sCD40L（2,4and 6μg/mL）for24,48 and 72h,respectively.The MTT assay was used to examine the effect of different concentrations of sCD40 Lon Leukemia HL-60 cell proliferation in vitro at different incubation times.RT-PCR was used to determine the effect of different concentrations of sCD40 Lon the expression of Survivin mRNA in HL-60 cells at different incubation times.ELISA was used to detect changes in cytokine IL-8levels in the culture supernatants of leukemia HL-60 cell of different concentrations of sCD40 Lat different incubation times.RESULTS Three different concentrations of sCD40 Lcould all significantly inhibit HL-60 cell proliferation and there was a dose-dependent tendency.There was statistical significance between different concentrations（F =150.076,P〈0.05）,and there was statistical significance between different time（F =358.969,P〈0.05）.sCD40 Lhad the best inhibitory effect on HL-60 for 48h.Three different concentrations of sCD40 L could all significantly inhibit the expression of Survivin mRNA in HL-60 cells.The 6.00μg/mL sCD40 Lgroup was the lowest.In the range of 24-72 h,Survivin mRNA expression in HL-60 cells was the lowest for 48 h.The expression rates of Survivin mRNA was respectively 89.7±6.2.Compared with As2O3 control group,it was statistically significant（P〈0.05）.Three different concentrations of sCD40 Lcould all significantly inhibit IL-8secretion in HL-60 cells.The6.00μg/mL group had the lowest expression rate.In the range of 24-72 h,IL-8secretion in HL60 cells was the lowest for 48 h.The expression rates of IL-8was respectively 380.2±29.5.Compared with

关 键 词：急性髓系白血病 可溶性CD40配体 HL-60 细胞增殖 SURVIVIN 白细胞介素8 

分 类 号：R733.71[医药卫生—肿瘤] R730.43[医药卫生—临床医学]