3D培养人工皮肤组织黑色素瘤模型建立及评价  

Establishment of three-dimensional skin reconstruction model of melanoma and its evaluation

作　　者：谢敏[1] 李明英[2] 张帆[2] 刘庆平[2] 秦建中[2] 迟彦[2,3] XIE Min;LI Mingying;ZHANG Fan;LIU Qingping;QIN Jianzhong;CHI Yan(Affiliated Zhongshan Hospital of Dalian University,Dalian 116001,China)

机构地区：[1]大连大学附属中山医院,辽宁大连116001 [2]大连大学生命科学与技术学院.糖脂代谢重点实验室 [3]辽宁师范大学生命科学学院

出　　处：《山东医药》2018年第25期27-30,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81241148)

摘　　要：目的建立体外人工皮肤组织黑色素瘤模型,为研究黑色素肿瘤侵袭和转移的机制及药物筛选奠定基础。方法采用3D培养技术,制备体外人工皮肤组织黑色素瘤C8161、A375模型。应用不同浓度血清(0.5%、5%)优化模型的最优培养条件。HE和免疫荧光观察该模型基底膜形成情况,检测黑色素瘤细胞C8161、A375侵袭能力。应用单层细胞培养检测细胞生长速度和细胞分泌E-钙黏素情况。采用Transwell实验检测不同黑色素瘤细胞侵袭能力。结果体外人工皮肤组织肿瘤模型建立成功,血清浓度0.5%为最佳3D培养条件。构建的人工皮肤组织肿瘤模型形成的基底膜良好,黑色素瘤细胞在此模型中发生侵袭和转移,且C8161侵袭能力强于A375细胞。侵袭力较弱的A375细胞增殖速度高于侵袭力较强的C8161细胞。C8161和A375细胞均不产生E-钙黏素。结论成功建立体外人工皮肤组织肿瘤模型,血清浓度为0.5%为最佳3D培养条件;黑色素瘤细胞的侵袭与生长速度并无直接关系,且失去E-钙黏素表达能力并不是导致肿瘤细胞侵袭的必要条件。Objective To establish the three-dimensional skin reconstruction model of melanoma,in order to lay a foundation for the study of mechanism of melanoma invasion and metastasis and drug screening. Methods Using the three-dimensional（ 3 D） culture technique to establish the three-dimensional skin reconstruction model of melanoma. Using different concentrations of serum to optimize the optimal culture conditions of the model. The formation of basement membrane and the ability of invasion of melanoma cells were tested by HE and immunofluorescence methods. The monolayer cell culture technique was used to test the cell growth rate and the secretion of E-cadherin. Transwell assay was used to detect the invasion of different melanoma cells. Results The three-dimensional skin reconstruction model of melanoma was successfully established. The serum concentration of 0. 5% was the best 3 D culture condition. The model formed the basement membrane in a good state. The invasion of melanoma cells was observed in this model,and the invasion ability of C8161 was stronger than that of A375 cells. The proliferation rate of A375 cells was higher than that of C8161 cells. Both C8161 and A375 cells did not produce E-cadherin. Conclusions The three-dimensional skin reconstruction model of melanoma is successfully established. The serum concentration of 0. 5% is the best 3 D culture condition. There is no direct relationship between the invasion of melanoma cells and the growth rate. The loss of E-cadherin expression ability is not a necessary condition for invasion of tumor cells.

关 键 词：恶性黑色素瘤 人工皮肤组织黑色素瘤模型 3D培养 

分 类 号：R739.5[医药卫生—肿瘤]