卵巢癌血管生成三维模型的体外构建  

Construction of a three-dimensional model of ovarian cancer angiogenesis in vitro

作　　者：周静雅 李欣[2] 韩庆 ZHOU Jingya;LI Xin;HAN Qing(Department of Obstetrics and Gynecology,Yichang Central People′s Hospital/the First College of Clinical Medical Science,Three Gorges University,Yichang,Hubei 443003,China;Department of Chemoradiotherapy,the Second People′s Hospital of Yichang/Hubei Key Laboratory of Tumor Microenvironment and Immunotherapy,Three Gorges University,Yichang,Hubei 443003,China)

机构地区：[1]三峡大学第一临床医学院/宜昌市中心人民医院妇产科,湖北宜昌443003 [2]宜昌市第二人民医院放化疗科/三峡大学肿瘤微环境与免疫治疗湖北省重点实验室,湖北宜昌443003

出　　处：《现代医药卫生》2023年第8期1275-1280,1287,共7页Journal of Modern Medicine & Health

基　　金：湖北省科技厅科学研究计划重点项目(D20211206);湖北省宜昌市科技攻关项目(A21-2-013);三峡大学肿瘤微环境与免疫治疗湖北省重点实验室开放基金项目(2017KZL02)。

摘　　要：目的利用人脐静脉内皮细胞(HUVECs)及卵巢癌患者腹水中原代卵巢癌细胞构建体外卵巢癌血管生成三维模型。方法2019年11月至2021年1月原代提取及培养HUVECs及卵巢癌细胞,分3组进行细胞培养。A组为无Matrigel基质胶条件下卵巢癌细胞和HUVECs间接共培养,B组为Matrigel基质胶条件下HUVECs单独培养,C组为Matrigel基质胶条件下卵巢癌细胞和HUVECs三维间接共培养。观察各组HUVECs血管生成情况。采用免疫印迹试验检测卵巢癌细胞条件培养液对HUVECs的CD105表达水平的影响,Transwell实验检测卵巢癌细胞条件培养液对HUVECs侵袭能力的影响。结果成功提取原代HUVECs和卵巢癌细胞,A组HUVECs有突起或伪足,但不形成管状或网状结构;B组HUVECs排列成网状,但没有细胞的变形、伪足和融合,也不形成类毛细血管样结构;C组HUVECs形成了不连续的三维类毛细血管样结构。卵巢癌细胞条件培养液可增强HUVECs的CD105表达并增强其侵袭能力。结论成功构建了体外卵巢癌血管生成三维模型,卵巢癌细胞具有促进HUVECs增殖及侵袭能力。该模型能为研究卵巢癌的血管生成提供一个更接近体内生理状态的体外模型。Objective To construct a 3D model of ovarian cancer angiogenesis in vitro using human umbilical vein endothelial cells(HUVECs)and protogenic ovarian cancer cells from ascites of ovarian cancer patients.Methods Primary HUVECs and ovarian cancer cells were extracted and cultured from November 2019 to January 2021,and cell culture was performed in three groups.In group A,ovarian cancer cells were indirectly co-cultured with HUVECs without Matrigel gel;in group B,HUVECs were separately cultured with Matrigel gel;and in group C,ovarian cancer cells and HUVECs were indirectly co-cultured with Matrigel gel.The angiogenesis of HUVECs in each group was observed.Western blotting was used to detect the effect of ovarian cancer cell conditioned medium on CD105 expression level in HUVECs,and Transwell assay was used to detect the effect of ovarian cancer cell conditioned medium on the invasion ability of HUVECs.Results Primary HUVECs and ovarian cancer cells were extracted successfully.HUVECs in group A had protuberance or pseudopodia,but did not form tubular or reticular structure.HUVECs in group B were arranged in a network,but there was no cell deformation,pseudopod and fusion,and no capillary-like structure.HUVECs in group C formed a discontinuous 3D capillary-like structure.Ovarian cancer cells conditioned medium enhanced CD105 expression and invasiveness in HUVECs.Conclusion The 3D model of ovarian cancer angiogenesis is successfully constructed in vitro,and the ovarian cancer cells have the ability to promote the proliferation and invasion of HUVECs.This model can provide an in vitro model for studying the angiogenesis of ovarian cancer which is more similar to the physiological state in vivo.

关 键 词：卵巢肿瘤 新生血管化 体外研究 三维模型 人脐静脉内皮细胞 

分 类 号：R737.31[医药卫生—肿瘤]