FOXQ1-siRNA对甲状腺乳头癌TPC-1细胞侵袭迁移的影响及其机制

Effects of FOXQ1-siRNA on invasion and migration of papillary thyroid carcinoma TPC-1 cells

目的探讨叉头框Q1(FOXQ1)-siRNA对甲状腺乳头状癌TPC-1细胞侵袭迁移能力的影响及其相关机制。方法通过LipofectamineTM2000将FOXQ1-siRNA和无意义序列siRNA分别转入TPC-1细胞,分别作为实验组和无义序列组,空白组不给予任何处理。转染成功后,利用qRT-PCR和Western blotting法分别检测FOXQ1 mRNA和蛋白的表达水平,采用Western blotting法检测TPC-1细胞中磷酸化蛋白激酶B(p-AKT)、基质金属蛋白酶7、9(MMP7、MMP9)的表达水平。转染48 h,采用Transwell小室试验观察细胞体外的侵袭、迁移能力。结果与空白组和无义序列组相比,实验组中FOXQ1 mRNA和蛋白的相对表达水平降低(P均<0.05);实验组中p-AKT、MMP7及MMP9蛋白的相对表达水平低于空白组和无义序列组(P均<0.05)。转染48 h,实验组中穿过小室膜的细胞数低于空白组和无义序列组(P均<0.05)。结论 FOXQ1-siRNA可抑制TPC-1细胞的侵袭迁移,这一作用可能是通过抑制PI3K/AKT信号通路实现。

Objective To investigate the effects of FOXQ1-siRNA on the invasion and migration of papillary thyroid carcinoma TPC-1 cells and its related mechanisms. Methods The FOXQ1-siRNA and non-sense sequence siRNA were transfected into the TPC-1 cells as the experimental and nonsense sequence group by lipofectamineTM2000,and the blank group did not receive any treatment. The expression of FOXQ1 mRNA and protein was detected by qRT-PCR and Western blotting,respectively. The expression of phosphorylated protein kinase B（ p-AKT）,matrix metalloproteinase 7,9（ MMP-7 and MMP-9） in the TPC-1 cells was detected by Western blotting. At 48 h,the invasion and migration of cells were observed by Transwell chamber assay. Results The relative expression levels of FOXQ1 mRNA and protein in the experimental group were significantly lower than those in the blank group and the nonsense sequence group,and the difference was statistically significant（ all P〈0. 05）. The relative expression levels of p-AKT,MMP-7,and MMP-9 in the experimental group were significantly lower than those in the blank group and the nonsense sequence group,and the difference was statistically significant（ all P〈0. 05）. At 48 h after transfection,the number of cells passing through the cell membrane in the experimental group was significantly less than that in the blank group and the nonsense sequence group,and significant difference was found（ all P〈0. 05）. Conclusion FOXQ1-siRNA inhibits the invasion and metastasis of TPC-1 cells by inhibiting the PI3 K/AKT signaling pathway.