摘要目的 研究非甾体抗炎药物吲哚美辛与化疗药物奥沙利铂对人肺癌A549细胞株裸鼠移植瘤微淋巴管生成的影响及化疗增敏作用.方法 将人肺癌A549细胞接种于裸鼠背部皮下,将裸鼠按随机数字表法分为4组:对照组、吲哚美辛组、奥沙利铂组、吲哚美辛和奥沙利铂联合用药组,并给予相应的药物.观察裸鼠生长情况,每7天测量1次肿瘤体积.42 d后处死裸鼠,切取移植瘤组织,用免疫组织化学染色法检测血管内皮生长因子C(VEGF-C)、β-连环素及生存素蛋白的表达,并测定微淋巴管密度,用实时荧光定量PCR检测VEGF-C mRNA及生存素mRNA的表达.用SPSS 16.0软件进行统计学分析,实验数据以((-x)±s)表示.多组间比较用方差分析,多组样本均数两两比较采用SNK检验,两变量的相关程度采用直线相关分析.结果 (1)吲哚美辛组、奥沙利铂组和联合用药组治疗后移植瘤体积分别为( 1322±327) mm3、(962 ±221) mm3和(611±161) mm3,低于对照组的(1664 ±318) mm3(F=23.33,P＜0.01)；(2)吲哚美辛组和联合用药组VEGF-C、生存素、β-连环素蛋白的表达及微淋巴管密度积分吸光度值均低于对照组(均P＜0.05).奥沙利铂组VEGF-C蛋白含量(20 825±2067)高于对照组(16 075±875,F=97.24,P＜0.05),β-连环素蛋白和微淋巴管密度( 17 396±1693,9666±978)与对照组(9824±1181,17 588±1698)比较无明显差异；(3) VEGF-C蛋白、生存素蛋白与微淋巴管密度呈直线正相关(r值分别为0.737和0.662,均P＜0.01),β-连环素蛋白与生存素蛋白呈直线正相关(r =0.582,P＜0.01),VEGF-C mRNA的表达与其蛋白表达呈直线正相关(r =0.873,P＜0.01).结论 吲哚美辛与奥沙利铂联合应用可提高抗肿瘤及抗微淋巴管生成作用,其作用机制可能是协同抑制了VEGF-C/VEGF受体3和Wnt信号途径.

abstractsObjective To investigate the antitumor lymphangiogenesis activity and the synergistic effect of indomethacin and oxaliplatin in a transplantation tumor model of human lung cancer in nude mice.Methods Human lung cancer A549 cells were injected subcutaneously into BALB/c nude mice.The mice were randomly divided into 4 groups:the control group,the indomethacin group,the oxaliplatin group,and the indomethacin combined with oxaliplatin group.The tumor volume was measured in a 7 day interval,and the mice were sacrificed after 42 days. The tumor tissues were collected and prepared for detection by immunohistochemistry of expressions of VEGF-C,Survivin,β-catenin and intratumoral microlymphatics density(MLD).Real-time fluorescence quantitative RT-PCR was performed for detecting the expression of VEGF-C and Survivin mRNA. SPSS version 16.0 was used for statistical analysis. Data were present as ((-x) ± s),and means were compared with analysis of variance.The correlation of 2 variables was analyzed by liner correlation analysis.Results The tumor volumes in the indomethacin group,oxaliplatin group,and indomethacin plus oxaliplatin group[ ( 1322 ±327) mm3,(962 ±221) mm3,(611 ± 161) mm3 ] decreased significantly as compared with that of the control group [ ( 1664 ± 318 ) mm3,F =23.331,P ＜ 0.01 ].Compared with the control group,the expression levels of VEGF-C,Survivin,β-catenin and MLD protein in the indomethacin group,and indomethacin plus oxaliplatin group were significantly reduced (P ＜ 0.05 ).Immunohistochemical studies showed an increase in the expression of VEGF-C in the oxaliplatin group (20 825 ± 2067 ) as compared to the control group ( 16 075 ± 875,F =97.24,P ＜ 0.05 ),but no significant differences in β-catenin and MLD ( 17 396 ± 1693,9666 ± 978) as compared with the control group (9824 ±1181,17 588 ± 1698)respectively,P ＞0.05.VEGF-C and Survivin proteins were positively correlated with MLD respectively (t =0.737,0.662,respectively P ＜ 0.01 ),and β-catenin protein was positively correlated with Survivin protein (r =0.582,P ＜ 0.01 ).VEGF-CmRNA was positively correlated with its protein (r =0.873,P ＜ 0.01 ).Conclusion The combined use of indomethacin and oxaliplatin promoted antitumor lymphangiogenesis activity,possible by inhibiting the VEGF-C/VEGFR-3 and Wnt pathways.