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- 2018
沉默Galectin-3基因对甲状腺癌细胞生长的影响
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Abstract:
目的:探讨沉默半乳凝集素-3(Galectin-3)基因表达对甲状腺癌细胞生长的影响。方法:采用RT-PCR及Western blot方法检测46例甲状腺癌及其相应的距癌灶1 cm以上癌旁正常组织中Galectin-3 mRNA和蛋白的表达。将阴性对照(NC)-siRNA、Galectin-3-siRNA转染人甲状腺癌SW579细胞,转染12、24、48 h后CCK-8法检测细胞增殖率; 转染48 h后,RT-PCR及Western blot检测Galectin-3 mRNA及蛋白的表达,流式细胞术检测凋亡率,Western blot法检测Ki67、PCNA、Cleaved caspase3、PI3K、AKT、p-AKT蛋白。以未转染细胞作空白对照。结果:甲状腺癌组织中Galectin-3 mRNA及蛋白的表达均高于癌旁正常组织(P<0.01)。与空白对照和NC-siRNA组比较,Galectin-3-siRNA组细胞Galectin-3 mRNA及蛋白表达降低(P<0.01); 细胞增殖率降低,凋亡率升高(P<0.01); Ki67、PI3K、PCNA和p-AKT蛋白表达降低,Cleaved caspase3蛋白表达升高(P<0.01)。结论:Galectin-3可能通过PI3K/AKT信号通路调控甲状腺癌细胞的生长。
Aim: To investigate the effect of silencing Galectin-3 gene on the growth of thyroid carcinoma cells.Methods: RT-PCR and Western blot methods were used to detect Galectin-3 mRNA and protein in 46 cases of thyroid carcinoma tissue and the corresponding normal tissue. Negative control(NC)-siRNA and Galectin-3-siRNA were transfected into human thyroid carcinoma SW579 cells after 12,24,48 h treatment, cell proliferation rate was detected by CCK-8 test; after 48 h, the Galectin-3 mRNA and protein were detected by RT-PCR and Western blot,respectively, apoptosis was detected by flow cytometry, and Ki67,PCNA, Cleaved caspase3, PI3K,AKT, and p-AKT proteins were detected by Western blot. The cells without transfection were the blank control.Results: The expressions of Galectin-3 mRNA and protein in thyroid carcinoma tissue were significantly higher than those in the corresponding normal tissue(P<0.01). Compared with those of the blank control and NC-siRNA group, the expression of Galectin-3 mRNA and protein in Galectin-3-siRNA group were significantly lower(P<0.01); the cell proliferation rate decreased and cell apoptosis rate increased(P<0.01); the expressions of Ki67, PCNA, PI3K, p-AKT protein decreased, and the expressions of Cleaved caspase3 protein increased(P<0.01).Conclusion: Galectin-3 may regulate the growth of thyroid carcinoma cells through PI3K/AKT signaling pathway
