白藜芦醇对人舌鳞状细胞癌系(Tca8113)缝隙连接蛋白43蛋白质表达和酪氨酸磷酸化的影响

The effect of resveratrol on the protein expression and tyrosine phosphorylation of Cx43 in human tongue squamous cell carcinoma line(Tca8113)

目的:探讨白藜芦醇(resveratrol,Res)对人舌鳞状细胞癌细胞(Tca8113)缝隙连接蛋白43(Connexin43,Cx43)蛋白表达和酪氨酸磷酸化的影响。方法:取体外培养的处于对数生长期的Tca8113细胞,根据Res的处理浓度分为3组,未加Res组(A组),Res 50μmol/L组(B组)和Res 100μmol/L组(C组)。采用RT-PCR和Western Blot分别测定Tca8113中Cx43 mRNA/蛋白表达水平和酪氨酸磷酸化水平;免疫组化染色测定细胞Cx43蛋白表达和定位。结果:RT-PCR结果显示Cx43 mRNA表达水平随着Res浓度升高而升高。Western Blot结果显示Cx43蛋白表达水平和酪酸磷酸化水平均显著升高;免疫组化染色显示Tca8113细胞株Cx43在细胞膜上的表达较少,主要位于细胞浆内。经Res处理的Tca8113细胞Cx43在细胞膜上的阳性细胞数明显增加,蛋白质表达和Cx43的磷酸化水平都较对照组显著增加(P <0. 05),且C组的高于B组(P <0. 01)。结论:Res可提高人Tca8113细胞株Cx43 mRNA/蛋白表达和酪氨酸磷酸化水平,从而调节Cx43在细胞膜和胞浆中的分布,可能具有抑制舌鳞癌细胞生长的作用。

Objective: To explore the effect of resveratrol on the protein expression and tyrosine phosphorylation of Cx43 in human tongue squamous cell carcinoma line( Tca8113). Methods: Tca8113 cells in logarithmic growth phase were divided into group A( control group,no resveratrol),group B( treated with 50 μmol/L resveratrol),group C( treated with 100μmol/L resveratrol). Western Blot was employed to analyze the protein expression and tyrosine phosphorylation of Cx43,and immunohistochemistrywas employed to analyze the protein content and position of Cx43. Results: The result of immunohistochemistryrevealed in Tca8113 cell line Cx43 was detected mainly either intracytoplas-mically or in plasma membrane free from contact with other cells. After treated with Res,Cx43 in plasma membrane was increased,and the protein content and tyrosine phosphorylation of Cx43 were both increased( P < 0. 05) and there was significant difference between group B and C( P < 0. 01). Conclusion: The protein content and tyrosine phosphorylation of Cx43 in Tca8113 can be increased by Res and the distribution of Cx43 in cell membrane and plasma can be regulated and then may inhibit the growth of human tongue squamous cancer.