摘要
目的探讨抑制甲状腺乳头状癌中斯钙素1(STC1)的表达对肿瘤细胞增殖和凋亡的影响及作用机制。方法将NC-siRNA和STC1-siRNA转染至甲状腺乳头状癌K1细胞,分别作为阴性对照组和STC1-siRNA组,以未转染的细胞作为空白对照组。蛋白质印迹法(Western blot)检测各组细胞中STC1、B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(BAX)、蛋白激酶B(AKT)、磷酸化的AKT(p-AKT)的相对表达量,CCK8检测细胞的增殖活力,流式细胞仪检测细胞的凋亡率及活性氧簇(ROS)水平。结果阴性对照组和空白对照组中STC1蛋白的相对表达量比较,差异无统计学意义(P>0.05);STC1-siRNA组中STC1蛋白的相对表达量明显低于空白对照组(P<0.01)。转染24、48、72 h后,STC1-siRNA组细胞的光密度(OD)均明显低于空白对照组(P<0.01);转染48 h后,STC1-siRNA组细胞的凋亡率明显高于空白对照组(P<0.01)。STC1-siRNA组中ROS水平及BAX蛋白的相对表达量均明显高于空白对照组,Bcl-2、p-AKT蛋白的相对表达量均明显低于空白对照组,差异均有统计学意义(P<0.01)。结论抑制甲状腺乳头状癌中STC1基因的表达可降低肿瘤细胞的增殖活力,诱导肿瘤细胞凋亡,其作用机制可能与下调AKT信号通路有关。
Objective To investigate the effect of inhibiting stanniocalcin 1(STC1) expression on the proliferation and apoptosis of papillary thyroid carcinoma cell, and to discuss the underlying mechanism. Method NC-siRNA and STC1-siRNA were transfected into papillary thyroid carcinoma K1 cells as negative control group and STC1-siRNA group, respectively, besides, the nontransfected cells were used as blank control group. Through Western blot, the relative expression of STC1, B-cell lymphoma/leukemia-2(Bcl-2), Bcl-2 associated X-protein(BAX), protein kinase B(AKT)and phosphorylated AKT(p-AKT) were detected, besides, CCK8 was utilized to evaluate the proliferative activity, and flow cytometry was applied to determine the apoptosis rate and reactive oxygen species(ROS) level. Result The relative expression of STC1 protein in the negative control group did not differ from that in the blank control group(P>0.05);while the relative expression of STC1 protein in the STC1-siRNA group was significantly lower than that in the blank control group(P<0.01). In 24, 48, 72 h after transfection, the optical density(OD) in STC1-siRNA group was markedly lower compared with that in blank control group(P<0.01); in 48 h after transfection, the cell apoptosis rate in STC1-siRNA group was higher than that in blank control group with statistical significance noted(P<0.01). In STC1-siRNA group,the ROS level and BAX protein expression were significantly increased, and that of Bcl-2 and p-AKT protein were significantly lower compared with blank control group, showing statistically significant difference(P<0.01). Conclusion Inhibition of STC1 gene expression in thyroid papillary carcinoma cancer results in decreased proliferative activity of tumor cells, inducing tumor cell apoptosis, of which the mechanism may be related to the down-regulation of AKT signaling pathway.
引文
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