TEM8在卵巢癌血管生成和侵袭迁移中的分子机制研究

The Molecular Mechanism of TEM8 in Angiogenesis,Invasion and Migration of Ovarian Cancer

目的:探讨肿瘤内皮细胞标志物8(tumor endothelial marker 8,TEM8)在卵巢癌血管生成和侵袭迁移中的分子机制。方法:体内实验中,选取4~5周龄SPF级BALB/C雌性小鼠20只,按随机原则分为两组,每组10只。分别将对照组细胞与实验组细胞接种于小鼠建立人卵巢癌皮下移植瘤模型。通过游标卡尺测量模型建立后各时期的肿瘤体积,2周后处死小鼠并分离皮下卵巢癌肿瘤组织并称重。通过反转录-聚合酶链反应(RT-PCR)观察各组组织中TEM8、血管内皮生长因子(vascular endothelial growth factor,VEGF)及CD34的表达水平。体外实验中,分别将处于对数生长期的人脐静脉内皮细胞(HUVEC)与转染了siRNA-TEM8的HUVEC细胞分为对照组及实验组,通过血管生成实验观察各组癌组织细胞的血管生成情况;通过Transwell试验观察各组癌组织细胞的侵袭迁移能力。结果:通过siRNA沉默TEM8后,实验组小鼠肿瘤体积及重量均小于对照组(P<0.05)。血管生成实验结果显示实验组小鼠血管生成数量明显少于对照组(P<0.05);Transwell试验显示,实验组迁移细胞数量与对照组相比较少(P<0.05);RT-PCR结果显示,实验组小鼠卵巢癌组织中TEM8、VEGF与CD34水平均明显低于对照组(P<0.05)。结论:TEM8能够促进卵巢癌血管生成,增强其侵袭迁移能力。

Objective:To investigate the molecular mechanism of tumor endothelial cell marker 8(TEM8)in angiogenesis,invasion and migration of ovarian cancer.Method:In vivo,twenty SPF BALB/C female mice aged from 4 to 5 weeks were randomly selected,according to the random principle,there were divided into two groups,10 cases in each group.Cells from the control group and the experimental group were inoculated into mice to establish human ovarian cancer subcutaneous transplanted tumor model.Tumor volume was measured by vernier caliper at each period after model establishment.Mice were sacrificed 2 weeks later and subcutaneous ovarian cancer tissues were isolated and weighed.The expression levels of TEM8,vascular endothelial growth factor(VEGF)and CD34 in each group were observed by reverse transcription-polymerase chain reaction(RT-PCR).In vitro,human umbilical vein endothelial cells(HUVEC)and siRNA-TEM8-transfected HUVEC cells were divided into control group and experimental group,respectively,and the angiogenesis of cancer tissue cells in each group was observed by angiogenesis assay.Transwell test was used to observe the invasion and migration of cancer cells in each group.Result:After TEM8 silencing by siRNA,the tumor volume and weight of mice in experimental group were smaller than those in control group(P<0.05).The results of angiogenesis experiment showed that the number of angiogenesis in experimental group was significantly less than that in control group(P<0.05).Transwell test showed that the number of migrating cells in the experimental group was less than that in the control group(P<0.05).The RT-PCR results showed that the levels of TEM8,VEGF and CD34 in ovarian cancer tissues of mice in experimental group were significantly lower than those in control group(P<0.05).Conclusion:TEM8 can promote angiogenesis of ovarian cancer and enhance its invasion and migration ability.