体外共培养hBMSCs和HUVECs对Panc-1胰腺癌细胞上皮间质转化的影响  

In vitro co culture of hBMSCs and HUVECs regulate epithelial mesenchymal transition of Panc-1 pancreatic cancer cells

作　　者：张兴隆 赵松[2] ZHANG Xing-long;ZHAO Song(Department of Oncology,The First Hospital of Handan City,Hebei 056000,China;Department of Pathology,Hebei Medical University,Shijiazhuang 056000,China)

机构地区：[1]河北省邯郸市第一医院肿瘤科,河北邯郸056000 [2]河北医科大学病理学教研室,河北石家庄050017

出　　处：《中国现代普通外科进展》2020年第2期93-98,共6页Chinese Journal of Current Advances in General Surgery

摘　　要：目的:利用三维共培养技术体外构建含多种细胞的胰腺癌微组织,研究骨髓间充质干细胞(hBMSCs)、内皮细胞(HUVECs)对Panc-1胰腺癌细胞上皮间质转化的影响。方法:水凝胶复合细胞培养构建三维胰腺癌微组织模型。第一组:单纯Panc-1细胞;第二组:Panc-1细胞复合HUVECs;第三组:Panc-1细胞复合hBMSCs;第四组:Panc-1细胞复合HUVECs和hBMSCs。实时定量PCR分析胰腺癌肿瘤侵袭相关的基质金属蛋白酶(MMP-9)、肿瘤上皮间质化E-cadherin和Snail基因。Western blot测定胰腺癌肿瘤侵袭相关的MMP-9、肿瘤上皮间质化E-cadherin和Snail蛋白。结果:HUVECs共培养不影响MMP-9、E-cadherin和Snail基因和蛋白的表达(P>0.05),hBMSCs共培养促进MMP-9和Snail基因和蛋白的表达、抑制E-cadherin基因和蛋白的表达(P<0.05),同时加入两种细胞促进MMP-9和Snail基因和蛋白的表达、抑制E-cadherin基因和蛋白的表达(P<0.05)。结论:利用水凝胶为载体,加入骨髓间充质干细胞、内皮细胞与Panc-1胰腺癌细胞共培养,成功构建复杂的胰腺癌肿瘤微组织。通过对胰腺癌上皮间质转化相关机制研究,发现骨髓间充质干细胞通过调节MMP-9和上皮间质化对胰腺癌的侵袭行为起重要作用。Objective:The aim of this study was to construct the microtissue containing multiple cells in vitro by three-dimensional co culture. Investigate the effects of co culture of hBMSCs and HUVECs regulate epithelial mesenchymal transition of Panc-1 pancreatic cancer cells. Methods: Construction of three dimensional microtissue model of pancreatic cancer by SIS hydrogel and cells. The first group: pure Panc-1 cells, the second group was Panc-1 cells combined with HUVECs, the third group was Panc-1 cells combined with hBMSCs, the fourth group: Panc-1 cells combined HUVECs and hBMSCs. Real-time quantitative PCR analysis of MMP-9, E-cadherin and Snail. Determination of MMP-9, E-cadherin and Snail protein by Western blot. Results: Real time quantitative PCR assay and Western Bolt results showed that the expression of Snail and MMP-9 gene and protein in Panc-1 cells was promoted by three different cell cultures in different degrees. The most significant effects were added the stem cells and endothelial cells simultaneously(P<0.05). Compared with the Panc-1 group, the expression of E-cadherin gene and protein in Panc-1 cells was inhibited by three different cells co culture in different degrees. The most significant effects were added the stem cells and endothelial cells simultaneously(P<0.05). Conclusions: The complicated microtissue of the pancreatic cancer was successfully constructed. hBMSCs play an important role in the invasion of pancreatic cancer by regulating MMP-9 and epithelial mesenchymal transition.

关 键 词：胰腺癌 肿瘤微环境 骨髓间充质干细胞 共培养 上皮间质转化 

分 类 号：R735.9[医药卫生—肿瘤] R730.23[医药卫生—临床医学]