摘要
目的探讨常用化疗药物紫杉醇(TAX)对人树突状细胞(DC)生物活性的影响及作用机制,为化疗联合生物免疫治疗抗肿瘤提供理论基础。方法体外实验:分别在紫杉醇5 ng/mL(b组)、10 ng/mL(c组)、20 ng/mL(d组)、40 ng/mL(e组)的浓度下及不加紫杉醇的条件下(a组)培养DC细胞,检测各组DC细胞的存活率、细胞成熟标志物及细胞因子分泌量,探究紫杉醇对人树突状细胞生物活性的影响,同时通过乳酸脱氢酶释放法(LDH)测定与紫杉醇处理的树突细胞共培养的细胞因子诱导的杀伤细胞(CIK)对肺癌A549细胞株的杀伤作用,测定紫杉醇对树突细胞抗原呈递作用的影响。体内实验:采用人肺癌细胞株A549的肿瘤组织块,皮下接种于裸鼠腋下,建立裸鼠肺癌模型。2 d后将30只裸鼠随机分成5组,分别为A组(对照组):细胞株A549+盐水(NS);B组:细胞株A549+DC+CIK;C组:细胞株A549+CIK+TAX;D组:细胞株A549+TAX;E组:细胞株A549+TAX+DC+CIK。每3天检测1次肿瘤大小、裸鼠存活时间。结果体外实验:在共培养6 d的前提下,b、c组第6天存活率与第0天存活率比较,差异无统计学意义(P>0.05),d、e组第6天存活率与第0天存活率比较,差异有高度统计学意义(P<0.01),e组存活率仅为17.5%。c组和e组CD86、CD40、MHCⅡ的表达以及IL-12的分泌与正常组相比有显著增加(P<0.05),d组除了CD86,其他检测指标较正常组差异均有统计学意义,b组中只有MHCⅡ较正常组差异有统计学意义,经10 ng/mL紫杉醇处理的细胞在效靶比5:1和10:1的情况下,杀伤效果明显高于未经紫杉醇处理的细胞。其中紫杉醇处理组的细胞在效靶比为10:1的情况下,杀伤活性已经到达(91.37±5.24)%。体内实验:B、C、D、E组的平均存活时间分别为33.0、34.0、31.8、41.8 d,明显高于对照A组(23.8 d)(P<0.05)。E组平均存活时间大于41.8 d,明显高于B、C、D组(P<0.01),其中E组中,45 d仍有3只存活,D组在前25 d体现出较好的治疗效果
Objective To explore the biological activity＇s effect and mechanism of common chemotherapy drug Paclitaxel （TAX） on human dendritic cells （DC）,and to provide a theoretical basis for the anti-tumor combination therapy of chemotherapy and immune therapy.Methods In vitro：DC were cultured separately without TAX （group a） and with different concentrations of TAX：5 ng/mL （group b）,10 ng/mL （group c）,20 ng/mL （group d）,and 40 ng/mL （group e）.In order to explore the effect of TAX on biological activity of DC,survival rates,cell maturation markers and cytokine secretion were detected.In order to explore the effect of TAX on DCs＇ antigen-presenting function,the cytotoxicity on the lung cancer cell line A549 of cytokine-induced killer cells （CIK）,which were co-cultured with pre-treated dendritic cells with TAX,was determined by lactate dehydrogenase release assay.In vivo：lung cancer model of nude mice was established by subcutaneously inoculating human lung cancer cell line A549 tumor tissue blocks into nude mice armpit.Two days later,30 mice were randomly divided into five groups：group A （control group）：cell line A549 ＋ saline （NS）,group B：cell line A549 ＋ DC ＋ CIK,group C：cell line A549 ＋ CIK ＋ TAX,group D：cell line A549 ＋ TAX,and group E：cell line A549 ＋ TAX ＋ DC ＋ CIK.The tumor size and the survival time of mice were examined every 3 days.Results In vitro：there was no statistical difference （P ＞ 0.05） between the cell survival rates in group b and group c on the day 6 and that on day 0 after co-cultured 6 days.There was significant statistical difference （P ＜ 0.01） between the cell survival rates in group d and group e on the day 6 and that on day 0 after co-cultured 6 days.The survival rate of DC in group e was only 17.5％.The expression of CD86,CD40 and MHC Ⅱ and the secretion of IL-12 in group c and group e increased significantly compared with control group.Those test results expect the expression of CD86 in group d were
出处
《中国医药导报》
CAS
2013年第33期15-19,共5页
China Medical Herald
基金
辽宁省科技厅
财政厅批准组建土木基础设施安全监测与控制等重点实验室(项目编号:辽科发[2011]20号)
关键词
紫杉醇
人树突状细胞
生物活性
肺癌
Paclitaxel
Human dendritic cells
Biological activity
Lung cancer
