新VEGFR2靶向抑制剂筛选及其抗肿瘤生物学活性检测

目的从8种新合成血管内皮细胞生长因子受体2（vascularendothelialgrowthfactorreceptor2，VEGFR2）靶向抑制剂中筛选有效抑制VEGFR2的药物，并检测其抗肿瘤效果。方法8种VEGFR2靶向抑制剂（标记为1—8号）各配成6种浓度分别作用于人脐静脉血管内皮细胞（humanumbilicalveinendothelialcells，HUVEC）和舌癌细胞株（Tea8113）细胞，同时设置5-氟尿嘧啶（5-FU）为阳性对照组，无药物组为空白对照组。用噻唑蓝（MTT）法检测细胞生长情况，免疫细胞化学法检测饱和浓度VEGFR2抑制剂组和空白对照组细胞VEGFR2表达，以验证抑制剂对VEGFR2的作用效应。结果与空白对照组相比，阳性对照组能显著抑制这两种细胞的生长增殖（P〈0．05），不同浓度各VEGFR2靶向抑制剂组对这两种细胞生长增殖的差异均无统计学意义（P〉0．05）。免疫细胞化学结果显示，空白对照组Tca8113细胞和HUVEC细胞膜上VEGFR2高表达；与空白对照组相比，饱和浓度不同抑制剂组的Tca8113细胞中只有6号试剂组可以显著下调细胞膜上VEGFR2表达（P=0．000），而HUVEC各组VEGFR2表达差异均无统计学意义（P〉0．05）。结论这些VEGFR2靶向抑制剂对人脐静脉血管内皮细胞和舌癌细胞生长增殖无抑制作用，其中只有6号抑制剂能显著下调舌癌细胞VEGFR2表达，可以作为进一步研究的候选药物。

Novel VEGFR2 targeted inhibitors: screening and detection of anti-tumor biological properties

Objective To screen effective medicine with VEGFR2 inhibitory activity from 8 novel VEGFR2 targeted inhibitors and to determine their anti-tumor properties. Methods Two cell lines（Tea8113 cells from tongue cancer and HUVEC ） were cultured respectively in groups with 6 different concentrations of the eight inhibitors （ marked No. I to No. 8 ）. Positive control using different concentrations of 5-FU and blank control without medicine were set up simultaneously. Cell growth was measured by MTr assay. Immunocytochemistry was carried out to deter- mined VEGFR2 levels in groups with different saturated inhibitors or in blank control. Results Cell growths of both cell lines were significantly inhibited in positive control groups comparing to that in blank control （ P 〈 0.05 ） ; there was no significant difference in cell growth between VEGFR2 inhibitor groups and blank control in the same cell line （P 〉0.05）. Immtmohistochemistry showed that VEGFR2 was highly expressed on the cell membranes of TcaS113 cells and HUVEC; comparing to blank control, only No. 6 VEGFR2 targeted inhibitor decreased VEGFR2 expression on Tca8113 cells significantly （P = 0. 000）, while all inhibitors showed no VEGFR2 inhibition on either cell line （P 〉 0.05）. Conclusion These VEGFR2 targeted inhibitors have no proliferative inhibition on the cell lines of HUVEC and TcaS113. However, No. 6 inhibitor decreases the VEGFR2 expression on Tca8113 cells significantly thus becom- ing a candidate for further study.