MV3细胞系体外构建恶性黑素瘤模型的研究  

Establishment of in vitro model for malignant melanoma with MV3 cells

作　　者：彭琳琳[1] 陆洪光[1] 

机构地区：[1]贵阳医学院附属医院皮肤科,550004

出　　处：《中华皮肤科杂志》2009年第2期97-100,共4页Chinese Journal of Dermatology

基　　金：基金项目：国家自然科学基金（30571685）

摘　　要：目的探讨MV3黑素瘤细胞接种至人去表皮的真皮（DED）上，体外构建皮肤恶性黑素瘤模型中黑素瘤细胞的侵袭方式。方法制备含部分基底膜成分的人去表皮的真皮组织（de-epidermized dermise，DED），利用PAS染色和Ⅳ型胶原免疫组化染色鉴定基底膜。将MV3黑素瘤细胞制成细胞瘤液，接种于DED表面，采用液下培养和空气一液面培养相结合的方式进行培养，用HE及免疫组化染色（S-100蛋白和HMB45），观察黑素瘤细胞在DED的分布。结果肉眼观察，DED在接种MV3黑素瘤细胞后颜色无明显变化。经PAS染色和Ⅳ型胶原免疫组化染色，验证DED中含有基底膜成分，分别位于DED表面和其内的皮肤附属器管壁上。组织学及免疫组化染色检查显示：MV3黑素瘤细胞在DED表面的基底膜带上生长分布，并在凹陷处形成大小不等团、灶；在皮肤附属器管壁内，呈环形、带状，贴附于基底膜上；在DED侧面，黑素瘤细胞弥散性浸润生长。组织中黑素瘤细胞S-100蛋白阳性和HMB45弱阳性。结论通过皮肤器官培养方式，可构建皮肤恶性黑素瘤的体外模型，基底膜影响恶性黑素瘤的侵袭生长方式。Objective To establish an in vitro model for malignant melanoma with a malignant melanoma cell line MV3 on de-epidermized dermis （DED） and to study the invasion mode of melanoma cells. Methods A human de-epidermized dermis was prepared with some elements of basal membrane （BM）. Then, the reconstructed BM was identified by periodic acid schiff （PAS） staining and immunochemical staining for collagen Ⅳ. MV3 ceils were seeded onto the prepared acellular dermis and maintained at the air-liquid interface for 13-15 days after 3-day submerged culture. Subsequently, the reconstructed malignant melanoma tissue was examined with hematoxylin and eosin （HE） staining and immunohistochemical staining with antibodies to S-100 protein and HMB45. Results No obvious changes were observed by naked eye in DED after the inoculation with MV3 cells. PAS staining and immunochemical staining for collagen Ⅳ confirmed the presence of BM component on the surface of DED and in the cavity of skin appendages in DED. Histological examination and immunochemical staining revealed that on the BM zone, MV3 cells grew into irregularly sized clusters; in the .cavity of skin appendages, they attached onto the BM and aggregated into circular or bandlike shape; and at the lateral side of DED, they invasively and diffusely grew, broke through the BM and intruded into the surrounding tissues of DED. The reconstructed tissue was positive for S-100 protein and weakly positive for HMB45. Conclusions The in vitro model of malignant melanoma could be reconstructed by skin organ culture system. And, the experiment suggests that BM could affect the invasive growth pattern of malignant melanoma cells.

关 键 词：黑色素瘤 真皮 基底膜 细胞 培养的 

分 类 号：R739.5[医药卫生—肿瘤] R651.15[医药卫生—临床医学]