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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。
The purpose of tumor angiogenesis and anti-tumor vascular drugs are there is a certain time window, in this time window, tumor neovascularization tends to normal morphology, the best anti-tumor effect. Therefore, this study was to evaluate the optimal time and molecular mechanism of recombinant human endostatin (rh-ES) on the normalization of tumor microvessels in mouse Lewis lung carcinoma (LLC) subcutaneous tumor models. Methods Forty LLC LLC xenografts were randomly divided into NS group and rh-ES group, with 20 mice in each group. NS group mice were intraperitoneally injected with saline (0.2 mL / d); rh-ES group mice were injected intraperitoneally with rh-ES [5 mg / (kg · d)]. Five mice were sacrificed on the 2nd, 4th, 6th and 9th day after treatment respectively. Tumor volume changes were measured in mice and tumor growth curves were plotted. At the same time, ELISA method was used to detect the correlation of tumor vascular remodeling, RGS5, VEGF and microvessel density, MVD). Results The C57 / BL6 mouse xenograft model was successfully established with a tumorigenic rate of 100%. The tumor volumes at day 4 and day 6 in rh-ES group were (0.81 ± 0.1) cm3 and (1.54 ± 0.4) cm3, respectively. The tumor volumes at day 4 and day 6 in NS group were (1.71 ± 0.2) cm3 and 0.4) cm3, the difference was statistically significant, P values were 0.04 and 0.03. The results of ELISA showed that the expression of RGS5 in rh-ES group was (4.02 ± 0.68) ng / mL and (2.98 ± 0.46) ng / mL on the 4th and 6th day, respectively, which was significantly lower than that in the NS group Respectively 0.01 and 0.02. The results of immunohistochemistry showed that the expression rates of VEGF in NS group were (44.10 ± 4.14)% and (45.13 ± 4.28)% on the 4th and 6th day, and those in rh-ES group were (27.16 ± 3.68)% and ± 3.08)% (n = 5), the expression of VEGF in rh-ES group was significantly decreased, with P values of 0.03 and 0.02, respectively. The MVD of NS group was 25.10 ± 5.28 and 25.68 ± 5.64 on the 4th and 6th day, respectively. The MVD of the rh-ES group was 15.50 ± 3.12 and 8.36 ± 2.04 respectively, and the rh-ES group was significantly reduced. The P values were 0.04 and 0.03 respectively. Conclusions rh-ES can be used as a window of normalization of blood vessels on the 4th and 6th days after Lewis lung cancer, which may be related to VEGF and MVD.