昆布多糖对人乳头状甲状腺癌细胞TPC-1生长信号转导的影响

Regulation of Laminarin on Growth Signal Transduction of Human Papillary Thyroid Cancer Cell Line TPC-1

目的探讨昆布多糖对人乳头状甲状腺癌细胞TPC-1生长信号转导影响研究。方法选用TPC-1人乳头状甲状腺癌细胞株,分别采用相应药物进行处理。采用噻唑兰(MTT)染色法检测昆布多糖对TPC-1细胞生长的抑制作用,采用流式细胞术分析昆布多糖对TPC-1细胞凋亡及细胞周期比例的影响,采用蛋白免疫印迹(Western Blot)法检测各组TPC-1细胞促甲状腺素受体(TSHR)、磷脂酶C-β3(PLC-β3)、蛋白激酶Cα(PKCα)蛋白表达水平。结果干预24h、48h、72h后,与对照组比较,昆布多糖0.4 g/L、0.8 g/L、1.6 g/L、2.0 g/L组TPC-1细胞生长抑制率较高,且昆布多糖0.4 g/L组<0.8 g/L组<1.6 g/L组及2.0 g/L组,差异有统计学意义(P<0.05)。与对照组比较,昆布多糖0.4 g/L、0.8 g/L、1.6 g/L组TPC-1细胞凋亡率较高,G2/M期细胞比例较高,且昆布多糖0.4 g/L组<0.8 g/L组<1.6 g/L组,差异有统计学意义(P<0.05)。与对照组比较,昆布多糖0.4 g/L、0.8 g/L、1.6 g/L组TSHR、PLC-β3、PKCα蛋白表达水平较低,且昆布多糖0.4 g/L组>0.8 g/L组>1.6 g/L组,差异有统计学意义(P<0.05)。结论昆布多糖能剂量依赖性抑制人乳头状甲状腺癌细胞TPC-1生长,使细胞阻滞于G2期,促进细胞凋亡,其作用可能与抑制TSHR-PLC-β3-PKCα生长信号通路转导有关。

Objective To analyzethe regulation of laminarin on growth signal transduction of human papillary thyroid cancer cell line TPC-1.Methods TPC-1 human papillary thyroid cancer cell lines were selected and treated with the corresponding drugs.The inhibitory effect of laminarin on growth of TPC-1 cells was detected by Thiazolyl Blue(MTT)staining method.The flow cytometry was used to analyze the effect of laminarin on the apoptosis and cell cycle ratio of TPC-1 cells.The expression levels of thyrotropin receptor(TSHR),phospholipase C-β3(PLC-β3),protein kinase Cα(PKCα)protein in TPC-1 cells of each group were detected by Western Blot method in TPC-1 cells of each group.Results After the intervention of 24 h,48 h and 32 h,compared with that of the control group,the growth inhibition rate of TPC-1 cells were higher in laminarin 0.4 g/L,0.8 g/L,1.6 g/L and 2.0 g/L groups,and laminarin group 0.4 g/L group<0.8 g/L group<1.6 g/L group and 2.0 g/L group,the differences were statistically significant(P<0.05).Compared with that of the control group,the apoptotic rate of TPC-1 cells was higher,the cell proportions in G2/M phase were higher in laminarin 0.4 g/L,0.8 g/L and 1.6 g/L groups,and laminarin 0.4 g/L group P<0.8 g/L group P<1.6 g/L group,the differences were statistically significant(P<0.05).Compared with those of the control group,the expression levels of TSHR,PLC-β3 and PKCαprotein were lower in laminarin 0.4 g/L,0.8 g/L and 1.6 g/L groups,and 0.4 g/L group>0.8 g/L group>1.6 g/L group,the differences were statistically significant(P<0.05).Conclusion Laminarin can inhibit the growth of human papillary thyroid cancer cell line TPC-1 in a dose-dependent manner,block cells in G2 phase and promote apoptosis.The effects may be related to the inhibition of TSHR-PLC-β3-PKCαgrowth signaling pathway transduction.