摘要
目的肿瘤血管生成和抗肿瘤血管药物均存在一定的时间窗,在该时间窗内,肿瘤新生血管形态结构趋于正常,抗肿瘤效果最佳。因此本研究评估重组人血管内皮抑素(rh-Endostatin,rh-ES)对小鼠Lewis肺癌(Lewis lung carcinoma,LLC)皮下移植瘤模型肿瘤微血管正常化的最佳时间及其分子机制。方法将40只LLC移植瘤模型小鼠随机分成NS组和rh-ES组,每组各20只。NS组小鼠腹腔注射生理盐水(0.2 mL/d);rh-ES组小鼠腹腔注射rh-ES〔5mg/(kg·d)〕。每组分别于治疗后的第2、4、6、9天处死5只小鼠留取标本。测量小鼠肿瘤体积变化,绘制肿瘤生长曲线。同时,ELISA方法检测肿瘤血管重构相关指标,G蛋白调节信号5(regulator of G-protein signaling 5,RGS5)、免疫组化检测血管内皮生长因子(vascular endothelial growth factor,VEGF)和微血管密度(microvessel density,MVD)。结果成功建立C57/BL6小鼠移植瘤模型,成瘤率为100%。rh-ES组第4和6天的肿瘤体积分别是(0.81±0.1)cm3和(1.54±0.4)cm3,NS组第4和6天的肿瘤体积分别是(1.71±0.2)cm3和(2.86±0.4)cm3,差异有统计学意义,P值分别是0.04和0.03。ELISA结果显示,rh-ES组肿瘤组织中RGS5的表达在治疗第4和6天分别是(4.02±0.68)ng/mL和(2.98±0.46)ng/mL,与NS组比较明显降低,P值分别是0.01和0.02。免疫组化结果表明,NS组第4和6天VEGF在肿瘤组织表达率分别为(44.10±4.14)%和(45.13±4.28)%,rh-ES组分别为(27.16±3.68)%和(25.08±3.08)%(n=5),rh-ES组VEGF表达率明显减少,P值分别是0.03和0.02。NS组第4和6天MVD分别为25.10±5.28和25.68±5.64,rh-ES组MVD分别为15.50±3.12和8.36±2.04,rh-ES组明显减少,P值分别是0.04和0.03。结论 rh-ES作用于Lewis肺癌后第4和6天可作为血管正常化时间窗,可能与VEGF和MVD相关。
OBJECTIVE Tumor angiogenesis present time window, anti angiogenic drugs also had a certain time window ,the vascular structure of tumor tended to be normal within time window , anti tumor effect was the best. The objective of this study was to analysis the optimal time course of recombinant human endostatin （rh-ES） to inhibit Lewis lung cancer （LLC） and relevant molecular mechanism in mice. METHODS The transplanted tumor model of LLC was established on C57/BL6 mice. Totally 40 LLC mice were randomly divided into NS group and rh-ES group （20 mice per group） by coin toss method. NS group experienced treatment of intraperitoneal injection for NS 0.2 mL/d, while rh-ES group was treated for rh-ES 5 mg/（kg·d）. The samples of 5 mice were obtained from d2 ,d4 ,d6 and d9 after treatment in NS group or rh-ES group, respectively. The tumor volume was calculated, tumor growth curve was mapped in mice. Regulator of G-protein signaling 5（RGS5）, vascular endothelial growth factor（VEGF） and microvessel density（MVD） in tumors were detected by ELISA and westernblotting. RESULTS The transplanted tumor model of LLC on C57/BL6 mice was established successfully. Rh-ES group tumor volume with d4 and d6 were （0.81±0. 1） and （1.54±0.4） cma （n=5）. NS group tumor volume with d4 and d6 were （1.71±0.2） and （2. 86±0.4） cma （n=5） , rh-ES group tumor volume less than NS group, the difference was statistically significant, P values were 0. 04 and 0. 03, respectively. ELISA showed that the expression of RGS5 in rh-ES group on d4 and d6 was （4.02±0.68） and （2.98±0.46） ng/mL （n=5）, compared with NS group （n=5）, P values were 0.01 and 0.02. The results of immunohistochemistry showed that the expression of VEGF in NS group on d4 and d6 was （44.10±4.14）%, （45.13 ^-4.28）~ （n = 5）. The expression of VEGF in rh-ES group on d4 and d6 was （27.16±3.68）%, （25.08±3.08）%（n=5）, the expression of VEGF in rh- ES group was significantly decreased, P values were 0.
出处
《中华肿瘤防治杂志》
CAS
北大核心
2016年第19期1291-1296,共6页
Chinese Journal of Cancer Prevention and Treatment
基金
2012四川省卫生厅科研项目(120284)
2013四川省卫生厅科研项目(130466)
2013南充市科技局应用技术与开发资金项目(13A0061)
2014年四川省科技厅项目(2014SZ0020-7)
2015年川北医学院博士启动基金(CBY15-QD09)
