胶质瘤转染Cx43cDNA与放疗的协同作用

The Synergistic Effect of Cx43cDNA Transfection and Radiotherapy on C6 Cells

目的探讨体外培养脑胶质瘤细胞转染Cx43cDNA与放疗是否存在协同作用及该作用与缝隙连接细胞间通讯功能(GJIC)的关系。方法LipofectAMINE2000介导质粒DNA转染,G418筛选,半定量RT-PCR及划痕荷载染料传输试验鉴定。分别以MTT法及流式细胞仪检测C6、C6-Non、C6-Cx43细胞放疗组和未放疗组的细胞增殖率及凋亡率,进而观察转染Cx43基因与放疗杀伤胶质瘤细胞是否有协同作用。结果①C6-Cx43细胞Cx43mRNA表达显著增加,染料传输能力较C6-Non明显增强;②转染Cx43cDNA,上调C6细胞GJIC功能后,放疗对脑胶质瘤细胞的毒性作用明显增强。结论①转染Cx43cDNA可上调C6细胞GJIC。②GJIC功能上调后,放疗对脑胶质瘤细胞的毒性作用明显增强,转染Cx43cDNA与放疗存在协同作用。③缝隙连接细胞间通讯可能是肿瘤转染Cx43cDNA与放疗存在协同作用的机制之一。

Objective To investigate the synergistic effect of Cx43cDNA transfection and radiotherapy on glioma and evaluate the relationship between the gap junctional intercellular communieation（GJIC） and the synergistic effect. Methods C6 cells were transfected by plasmid DNA mediated by LipofectAMINE 2 000. The transfected cells were obtained by using G418.Then transfected cells were identified by RT-PCR and scrape loading dye transfer test. The proliferative and apoptosis rates of C6 cells were determined respectively by MTT and flow cytometry. The synergistic effect of Cx43 DNA transfection and radiotherapy on C6 cells was observed. Results ①The level of Cx43mRNA was significantly elevated in C6-Cx43 group compared to that in C6-Non group, and the dye transfer ability of C6-Cx43 was stronger than that of C6-Non. ②After transfected Cx43cDNA, the GJIC function of C6-Cx43 was significantly upregulated, and the cytotoxicity effect of radiotherapy on C6 cells was amplified. Conclusions ①Cx43cDNA transfection can elevate the level of Cx43mRNA in cells, and upregulate GJIC function of C6 cells. ②The upregulation of GJIC function can significantly amplify the cytotoxicity effect of radiotherapy on C6 cells, and there is the synergistic effect of Cx43cDNA transfection and radiotherapy on C6 cells. ③ It is suggested that the upregulation of GJIC function may be one of the mechanism of the synergistic effect of Cx43cDNA transfection and radiotherapy on C6 cells.