腺样囊性癌细胞与大鼠施万细胞共培养的实验研究

目的 ：建立腺样囊性癌细胞与大鼠施万细胞共培养模型，探讨细胞间相互作用时BDNF、TrkB和E-cadherin的表达变化情况。方法 ：采用Transwell系统建立腺样囊性癌细胞与大鼠施万细胞共培养模型，共培养96 h后，观察肿瘤细胞的形态学变化，以ELISA法检测各组培养液中BDNF含量，实时定量PCR（RT-PCR）、Western免疫印迹检测肿瘤细胞中BDNF、TrkB及E-cadherin的表达情况。采用SPSS17.0软件包对实验结果进行统计学处理。结果 ：与施万细胞共培养的腺样囊性癌细胞发生长梭形及多角形改变，E-cadherin表达下调，TrkB表达上调，BDNF表达未见明显变化。与腺样囊性癌细胞系共培养的培养液中，施万细胞表达BDNF的量比单独培养的施万细胞显著增高；而对照组黏液表皮样癌细胞与施万细胞共培养时未发生类似改变。结论 ：BDNF/TrkB信号通路在唾液腺腺样囊性癌嗜神经侵袭过程中发挥重要作用，但其具体分子机制有待进一步研究。

Research on the co-culture model of salivary adenoid cystic carcinoma cells and rat schwann cells

PURPOSE : To establish a co-culture system of salivary adenoid cystic carcinoma (SACC) cells with rat schwann cells for further investigating the changes of brain-derived neurotrophic factor (BDNF), tropomysin-related kinase B (TrkB) and E-cadherin after cells interaction. METHODS : Transwell system was used to establish co-culture model of SACC cells with rat schwann cells. After co-culture for 96 hours, the morphology of tumor cells were investigated, the change of BDNF in co-culture medium of each group was tested by enzyme-linked immunosorbent assay (ELISA) and the expression of BDNF, TrkB and E-cadherin in tumor cells were detected by RT-PCR and Western blot. The date was analyzed using SPSS 17.0 software package. RESULTS : Compared with the single culture group, the morphology of SACC cells co-cultured with schwann cells changed to spindle-shape and polygon-shape; the expression of E-cadherin in co-cultured SACC cells was down-regulated while the expression of TrkB and phospho-TrkB was up-regulated, but the expression of BDNF had no obvious change; more expression of BDNF was detected in the co-culture medium secreted by schwann cells. The mucoepidermoid carcinoma (MEC) cells in the control group, co-cultured with schwann cells, had no similar changes. CONCLUSIONS : BDNF/TrkB signal pathway plays an important role in the process of perineural invasion of SACC, but the concrete molecular mechanism needs further experimental investigation.