下调Galectin-3基因表达抑制神经母细胞瘤增殖及诱导凋亡的机制

Study on the mechanism of down - regulation of Galectin - 3 gene expression on proliferation and apoptosis of neuroblastoma

目的观察下调半乳糖凝集素-3（Galectin-3）基因表达对神经母细胞瘤增殖及凋亡的影响。方法反转录-聚合酶链反应（RT-PCR）及Westernblot分别检测Galectin-3基因在神经母细胞瘤组织的mRNA及蛋白表达；Galectin-3-siRNA（小干扰RNA）转染人神经母细胞瘤细胞株SK-N-SH，另设阴性对照组（NC-siRNA）和空白对照组（Contr01），细胞转染48h后，Westernblot检测各组细胞中Galectin-3、活化的含半胱氨酸的天冬氨酸蛋白水解酶-3（CleavedCaspase-3）、磷脂酰肌醇3-激酶（P13K）、磷酸化的丝氨酸苏氨酸激酶（Akt）蛋白表达；细胞计数试剂盒（CCK-8）检测细胞增殖；流式细胞仪检测细胞凋亡。结果Galectin-3基因在神经母细胞瘤中的mRNA及蛋白表达均显著高于瘤旁组织（tmRNA=8．174，PmRNA=0．001；t蛋自=17．648，P蛋白=0．000）；转染Galectin-3-siRNA后的SK-N-SH细胞中Galectin-3的蛋白表达显著低于Control组（t=12．461，P=0．000）。结论下调Galectin-3基因表达可显著抑制神经母细胞瘤增殖及诱导细胞凋亡，其机制与抑制P13K／Akt信号通路有关。

Objective To investigate the effect of downregulation of Galectin - 3 gene expression on proliferation and apoptosis of neuroblastoma. Methods The mRNA and protein expression of Galectin - 3 in neuroblastoma tissues were detected by reverse transcriptase - polymerase chain reaction （ RT - PCR） and Western blotting; Galectin -3 -siRNA （small interfering RNA） was transfected into human neuroblastoma cell line SK -N -SH, a negative control group （NC -siRNA） and control group （Control） were set, ex- pression of Galectin - 3, Cleaved Caspase -3, phosphatidylinositol 3 kinase （ PI3K）, phosphorylated pro- tein kinase B （p -Akt） protein were detected by Western blotting cells were transfected for 48 h; cell pro- liferation was detected by cell counting kit - 8 （ CCK - 8） ; cell apoptosis were determined by flow cytome- try. Results The mRNA and proteinexpression of Galeetin - 3 gene in neuroblastoma significantly higher than the tumor adjacent tissues （tmRNA=8. 174, PmRNA =0. 001 ; t蛋白= 17. 648, P蛋白 =0. 000） ; the ex- pression of Galectin - 3 proteins transfected Galectin - 3 - siRNA in the SK - N - SH cells was lower com- pared with Control group （ t = 12. 461, P=0. 000）. Conclusion Downregulation of Galectin - 3 gene ex- pression can significantly inhibit proliferation and induce apoptosis of neuroblastoma, and the mechanism is related to the inhibition of PI3K/Akt signaling pathway.