血管内皮细胞条件培养基促进胃癌细胞增殖和迁移  

Vascular endothelial cell conditioned medium promotes the proliferation and migration of gastric cancer cells

作　　者：余晓刚 朱晓丹 周法庭[1,2] 孙志民 郭庆[1,2] 张蓉[1,2] YU Xiaogang;ZHU Xiaodan;ZHOU Fating;SUN Zhimin;GUO Qing;ZHANG Rong(The First College of Clinical Medical Science,China Three Gorges University,Yichang 443003,Hubei,China;Yichang Central People's Hospital,Yichang 443003,Hubei,China;The Second People's Hospital of Yichang,Yichang 443000,Hubei,China)

机构地区：[1]三峡大学第一临床医学院,湖北宜昌443003 [2]宜昌市中心人民医院,湖北宜昌443003 [3]宜昌市第二人民医院,湖北宜昌443000

出　　处：《西部医学》2023年第7期970-975,981,共7页Medical Journal of West China

基　　金：宜昌市卫生科研资助项目(A19-301-02)。

摘　　要：目的 探讨血管内皮细胞培养基对胃癌细胞增殖和迁移的影响,分析血管内皮细胞调控胃癌细胞发生转移的机制。方法 设置对照组和共培养组,分别将正常培养基和人脐静脉血管内皮细胞HUVEC条件培养基作用于胃癌细胞HGC27,采用MTT法和划痕试验检测胃癌细胞HGC27的增殖活性和迁移能力。设置Control组、6 h组、12 h组和24 h组,以HUVEC的条件培养基作用于胃癌细胞6、12和24 h,Western blot检测EMT标志物和紧密连接蛋白的表达变化,激光共聚焦显微镜观察细胞骨架和紧密连接蛋白的分布变化。结果 MTT及划痕试验表明HUVEC条件培养基促进胃癌细胞HGC27的增殖和迁移。与Control组比较,间接共培养后胃癌细胞的形态呈间充质状改变,丝状伪足样凸起数量显著增加。Western blot结果显示间接共培养后胃癌细胞上皮标志物E-cadherin表达水平逐渐下降,而间充质标志物N-cadherin和MMP-9逐渐增加,具有时序性变化规律。ZO-1和Occludin的表达也逐渐下降,细胞膜分布减少。结论 间接共培养下,血管内皮细胞通过上调胃癌细胞MMP-9,破坏紧密连接,促进胃癌细胞增殖和迁移。Objective This study was to investigate the effect of vascular endothelial cell conditioned medium on the proliferation and migration of gastric cancer cells,and further to explore the potential mechanism of gastric cancer metastasis.Methods Control group and co-culture group were set up.Gastric cancer cells HGC27 were stimulated with human umbilical vein endothelial cells(HUVEC)conditioned mediumr or normal medium,respectively,and the proliferation and migration of HGC27 were detected by MTT assay and scratch test,respectively.Then control group,6h group,12h group and 24h group were built the following tests.The expression of epithelial-mesenchymal transition(EMT)and tight junction were detected by Western blot after HGC27 treated with HUVEC conditioned medium for 0h,6h,12h and 24h durations.Furthermore,the distribution changes of cytoskeleton and tight junctions were observed with laser confocal microscopy.Results MTT and scratch test showed that HUVEC medium promoted the proliferation and migration of HGC27,respectively.Compared with the Control group,the morphology of HGC27 turned to mesenchymal after co-culture,and the number of filamentous pseudopod-like protrusions(FLPs)increased significantly.WB results showed that the expression level of E-cadherin decreased gradually,while N-cadherin and MMP-9 increased gradually in a time-depend manner.The expression of ZO-1 and Occludin also displayed a decreasing trend,and the distribution were both decreased obviously.Conclusion Vascular endothelial cells promote gastric cancer cell migration by up-regulating MMP-9 and destroying tight junctions in the indirect co-culture condition.

关 键 词：胃癌 上皮间充质转化 血管内皮细胞 基质金属蛋白酶 紧密连接 

分 类 号：R735[医药卫生—肿瘤]