阻断VEGFR-3表达对肿瘤细胞诱导的淋巴管内皮细胞增殖的影响

Effect of blocking the expression of VEGFR-3 on lymphatic endothelial cell proliferation in a tumor cell line

目的观察阻断VEGFR-3(Flt 4)表达对肿瘤细胞(前列腺癌细胞株PC3)诱导的淋巴管内皮细胞增殖的影响.方法实验分4组,第1组为对照组.每组有6孔,每孔具有相同细胞数2×105/ml,实验组每孔各加入试剂100 ml/L.第1组(对照组)加入淋巴管内皮细胞完全条件培养液(LEC)1 ml,第2组加入LEC 1 ml+兔血清100 μl,第3组加入LEC 1 ml+PC3细胞上清100 μl.第4组加入LEC+抗Flt 4抗体100 μl.并于24、48、72、96 h观察各组淋巴管内皮细胞生长情况.各时间段计数第1～3组细胞数,第4组于72 h计数后,去除抗体,重新加入LEC+PC3细胞上清继续培养至120 h;除96 h外,其余各时间段均计数细胞数.比较各组细胞增殖情况.结果第1、2组淋巴管内皮细胞在加试剂后各时间段两组细胞数及形态无明显差别,第3组加入PC3细胞上清后,细胞数明显多于第1、2组.第4组加试剂后24、48、72 h细胞计数均少于前24 h.清除抗体后加入PC3细胞上清48 h计数细胞,细胞数仅略见增加.结论 VEGF-C高表达的PC3细胞上清能显著刺激淋巴管内皮细胞增殖,阻断Flt4表达,可在一定程度上阻断PC3细胞上清促淋巴管内皮细胞增殖作用.

Purpose To observe the effect of blocking the expression of VEGFR-3 on lymphatic endothelial cells proliferation in a tumor cell line PC3. Methods There were four groups in this experiment： one control and three experimental groups, 6 pores in each group. The number of lymphatic endothelial cells（LEC） in each pore was equal. LEC was added into the first group, LEC and rabbit serum into the second group, LEC and superuatant of PC3 into the third group, and LEC and the antibody to VEGFR-3 into the fourth group. We observed the growth and proliferation of LEC in every group after adding reagents at 24h, 48h, 72h and 96 h. Cell counting was performed from the first group to the third group at every point of time. The antibody was cleared at 72 hours after adding reagent, and the superuatant of PC3 ＋ LEC were added into the fourth group. Cells of the fourth group were cultured till 120h and counted at 120h. Cell counting was done at every point of time to compare theoproliferation of cells in each group except 96 h. Results After adding reagents into the lymphatic endothelial cells, there were no difference in shape and growth between the control and the second group at 24 h, 48 h, 72 h and 96 h. The cells of the third group grew actively, and the number of cells was higher than that of the first and the second groups. In the fourth group, the number of cells was lower than those at 24 h, 48 h and 72 h. The number of cells was increased a little at 48h after adding the superuatant of PC3 cell. Conclusions The superuatant of cultured PC3 cells promote lymphatic endothelial cell proliferation obviously. If the expression of Fit 4 is blocked, the proliferation of lymphatic endothelial cells induced by superuatant of PC3 cell culture will be blocked in some degree.