鱼藤素对胆管癌TFK-1细胞诱导肿瘤新生血管生成的抑制作用

摘要

目的 观察并评价鱼藤素在抑制胆管癌细胞诱导人脐静脉内皮细胞(HUVECs)增殖、迁移以及体内血管生成中的作用.方法 以CCK-8检测鱼藤素对HUVECs增殖的影响;Ki-67检测鱼藤素对TFK-1细胞诱导HUVECs增殖能力的影响;Transwell细胞迁移实验以及小管形成实验检测鱼藤素对TFK-1细胞诱导HUVECs迁移和体外血管生成能力的影响;基质胶血管生成实验检测鱼藤素对TFK-1细胞在肿瘤新生血管生成能力的影响.结果 鱼藤素在体外对HUVECs活性无影响;添加胆管癌TFK-1细胞培养上清24 h后HUVECs Ki-67表达阳性率为(97.3±0.71)％,普通1640培养基24h后HUVECs Ki-67表达阳性率为(77.22±1.83)％,差异有统计学意义(P＜0.01);添加鱼藤素1 μmol/L培养24 h后Ki-67表达阳性率为(87.86±2.81)％,与加入TFK-1上清组(97.3 ±0.71)％比较差异有统计学意义(P＜0.05);将1μmol/L鱼藤素加入含有胆管癌TFK-1细胞上清的HU-VECs作用24h后,对照组穿膜细胞数为(236.7±13.86)个/视野,鱼藤素1μmol/L组为(94.67±7.45)个/视野,两组比较差异有统计学意义(P＜0.01);将1 μmol/L鱼藤素加入含有胆管癌TFK-1细胞上清的HUVECs作用24 h后,对照组管腔样结构为(60.67±6.12)个/视野,鱼藤素1μmol/L组为(24.33±3.53)个/视野,两组比较差异有统计学意义(P＜0.01);鱼藤素组基质胶栓瘤体中新生血管数量为(9.60±1.36)个/视野,对照组为(4.49±0.93)个/视野,两组比较差异有统计学意义(P＜0.05);CD31和血管内皮生长因子的表达亦明显低于对照组.结论 鱼藤素能够明显抑制胆管癌TFK-1细胞诱导人脐静脉内皮细胞在瘤体中新生血管生成的能力.%Objective To observe and evaluate the inhibitory effects of deguelin on cholangiocarcinoma cells induced human umbilical vein endothelial cells(HUVECs) proliferation,migration and in vivo vascular generation.Methods The effects of deguelin to HUVECs on proliferation were detected by CCK-8.The effects of deguelin to cholangiocarcinoma TFK-1 cells inducing HUVECs on proliferation were detected by Ki-67.The migration and tube formation ability of HUVECs inducing by TFK-1 cells were determined by transwell assay and tube formation assay respectively.The effects of deguelin on TFK-1 cells to angiogenesis in vivo were examined by Matrigel plug assay.Results The deguelin had no effects to HUVECs on proliferation.The expression of Ki-67 of HUVECs treated by TFK-1 cells medium for 24 h was(97.3 ±0.71) ％,comparing to(77.22 ± 1.83) ％ in 1640 medium,the difference was significant (P ＜ 0.01),and (87.86 ± 2.81) ％ in TFK-1 cells medium with addition of deguelin (P ＜ 0.05).The invaded HUVECs that treated by deguelin 1 μmol/L in TFK-1 cells medium for 24 h was (94.67 ± 7.45),comparing with(236.7 ± 13.86) in control group,the difference was significant(P ＜0.01).The number of new tubules that treated by deguelin 1 μmol/L in TFK-1 cells medium for 24 h was 24.33 ± 3.53,comparing with 60.67 ± 6.12 in control group,the difference was significant (P ＜ 0.01).The number of new vessels in Matrigel plug that treated by deguelin group was 9.60 ± 1.36,comparing with 4.49 ± 0.93 in control group,the difference was significant(P ＜ 0.05),and the expression of CD31 and VEGF was inhibited by deguelin.Conclusions Deguelin could inhibit the angiogenesis of human umbilical vein endothelial cells that induced by cholangiocarcinoma TFK-1 cells.