论文部分内容阅读
目的研究罗勒多糖对人淋巴管内皮细胞(HLECs)的血管内皮细胞生长因子受体-2/3(VEGFR-2/3)表达的影响,揭示罗勒多糖抗肿瘤转移的分子机制。方法乏氧条件下体外培养HLECs,实验分为3组,A组为不加罗勒多糖处理的空白对照组,B组(罗勒多糖,200μg·m L~(-1)),C组(罗勒多糖,400μg·m L~(-1)),观察各组HLECs体外成管能力;实时荧光定量PCR检测罗勒多糖处理的HLECs中VEGFR-2/3 mRNA的表达;免疫细胞化学分析VEGFR-2/3蛋白的表达差异。结果 HLECs体外成管数目分别为:29.6±5.47(A组),23.6±3.68(B组),19.2±2.00(C组);与A组比较,B、C组均能减少HLECs的体外成管数目(P<0.05)。VEGFR-2 mRNA相对表达量分别为:1(A组),0.59±0.25(B组),0.90±0.13(C组);与A组比较,B、C组VEGFR-2 mRNA相对表达量下降,但差异没有统计学意义。VEGFR-3 mRNA相对表达量分别为:1(A组),0.52±0.19(B组),0.19±0.09(C组);与A组比较,B、C组VEGFR-3 mRNA相对表达量均下降,差异有统计学意义(P<0.05)。VEGFR-2蛋白表达累积光密度分别为:2.51±0.03(A组),2.42±0.03(B组),2.12±0.03(C组);与A组比较,B、C组VEGFR-2 mRNA相对表达量下降,但差异没有统计学意义。VEGFR-3蛋白表达累积光密度分别为:3.72±0.28(A组),2.91±0.26(B组),2.82±0.20(C组);与A组比较,B、C组VEGFR-3蛋白相对表达量均下降,差异有统计学意义(P<0.05)。结论罗勒多糖可降低HLECs体外成管能力和并显著下调VEGFR-3的表达,这可能是罗勒多糖抑制淋巴管新生,抗肿瘤转移的分子机制。
Objective To study the effect of basil polysaccharide on the expression of vascular endothelial growth factor receptor-2/3 (VEGFR-2/3) in human lymphatic endothelial cells (HLECs) and reveal the molecular mechanism of basil antitumor metastasis. Methods HLECs were cultured in vitro under hypoxic conditions. The experiment was divided into three groups. Group A was basal control without basilar treatment, group B (basilar polysaccharide, 200μg · m L -1), group C , 400μg · m L -1). The ability of in vitro tube formation of HLECs in each group was observed. The expression of VEGFR-2/3 mRNA in HLECs treated with basilar polysaccharide was detected by real-time fluorescence quantitative PCR. The expressions of VEGFR-2/3 Differences in protein expression. Results The numbers of in vitro tubes of HLECs were 29.6 ± 5.47 (group A), 23.6 ± 3.68 (group B) and 19.2 ± 2.00 (group C) respectively. Compared with group A, B and C groups could reduce the in vitro tube formation of HLECs Number (P <0.05). Compared with group A, the relative expression of VEGFR-2 mRNA in group B and group C was lower than that in group A (P <0.05) But the difference was not statistically significant. The relative expression levels of VEGFR-3 mRNA in group B and C were respectively 1 (group A), 0.52 ± 0.19 (group B) and 0.19 ± 0.09 (group C) , The difference was statistically significant (P <0.05). The cumulative optical density of VEGFR-2 protein expression was 2.51 ± 0.03 (group A), 2.42 ± 0.03 (group B) and 2.12 ± 0.03 (group C) respectively. Compared with group A, VEGFR- The amount decreased, but the difference was not statistically significant. The cumulative optical density of VEGFR-3 protein expression was 3.72 ± 0.28 (group A), 2.91 ± 0.26 (group B) and 2.82 ± 0.20 (group C) respectively. Compared with group A, VEGFR- Volume decreased, the difference was statistically significant (P <0.05). Conclusion Basil polysaccharide can reduce the tube formation ability of HLECs in vitro and significantly down-regulate the expression of VEGFR-3, which may be the molecular mechanism of basil inhibition of lymphangiogenesis and anti-tumor metastasis.