维甲酸对睾丸癌细胞缝隙连接蛋白Cx43表达及其功能的影响

目的 探讨维甲酸对睾丸癌细胞缝隙连接蛋白（Cx43）表达及其功能的影响。方法 体外培养的睾丸癌I-10细胞分别予2.5、5.0、10.0 μmol/L维甲酸处理24 h，Western blot法检测I-10细胞Cx43蛋白表达，细胞免疫荧光法观察细胞膜表面Cx43蛋白分布的改变，细胞接种荧光示踪法检测细胞荧光传递功能的变化。结果 Western blot结果显示，与对照组比较，维甲酸2.5、5.0、10.0 μmol/L可增加I-10细胞中Cx43蛋白表达(P<0.05)，其增强率分别为43.14%±2.1%、58.09%±1.8%、143.13%±1.6%；细胞免疫荧光法结果显示，维甲酸2.5、5.0、10.0 μmol/L可显著增加I-10细胞膜表面Cx43蛋白的表达；细胞接种荧光示踪法结果显示，与对照组比较，维甲酸2.5、5.0、10.0 μmol/L可增强I-10细胞间荧光传递功能(P<0.01)，增强率分别为26.1%±2.3%、63.3%±1.6%、140.5%±3.4%。结论 维甲酸可通过增加I-10细胞Cx43蛋白的表达来增强细胞间的缝隙连接功能。

Effects of Retinoic Acid on the Expression of Cx43 and Its Gap Juncion Intercellular Communication Function

Objective To investigate the effects of retinoic acid (RA) on the expression of Cx43 and its gap junction intercellular communication function in testicular cancer cells. Methods Cultured testicular cancer cells I-10 were treated with different concentration of RA (2.5,5.0,10.0 μmol/L). The expression of Cx43 in I-10 cells was detected by Western blot, and the distribution and location of Cx43 on cellular membrane was studied with immunofluorescence assay. Parachute assay was used to detect the function of gap junction intercellular communication composed of Cx43 in I-10 cells. Results RA (2.5, 5.0, 10.0 μmol/L) markedly increased the expression of Cx43 in I-10 cells, the enhancement ratios were 43.14%±2.1%, 58.09%±1.8%, 143.13%±1.6%, respectively. The result of immunofluorescence assay showed that RA (2.5, 5.0, 10.0 μmol/L) obviously increased the level of Cx43 located on the cellular membrane of I-10 cells. The result of parachute assay demonstrated that RA (2.5,5.0,10.0 μmol/L) could enhance the intercellular dye coupling through gap junction, the enhancement ratios were 26.1%±2.3%, 63.3%±1.6%, 140.5%±3.4%, respectively. Conclusion RA could enhance the gap junction intercellular communication by increasing the expression of Cx43 in I-10 cells.