沉默CITED1基因对甲状腺乳头状癌K1细胞生物学特性的影响

Effects of CITED1 gene silencing on biological functions of papillary thyroid carcinoma K1 cells

目的:探讨沉默CITED1(Cbp/p300 interacting transactivator with Glu/Asp rich carboxy-terminal domain 1)基因表达对甲状腺乳头状癌K1细胞增殖、凋亡和迁移等生物学行为的影响。方法:将携带特异性靶向CITED1基因的shRNA(CITED1-shRNA)及其阴性对照shRNA(negative control-shRNA,NC-shRNA)的重组慢病毒分别感染甲状腺乳头状癌K1细胞株,采用实时荧光定量PCR法和蛋白质印迹法检测CITED1基因沉默效率;然后采用CCK-8法检测细胞增殖,FCM法检测细胞周期和细胞凋亡,划痕愈合实验检测细胞迁移能力。结果:经携带CITED1-shRNA的重组慢病毒感染后,甲状腺乳头状癌K1细胞中CITED1 mRNA和蛋白的表达水平均明显降低(P值均<0.01),证明成功获得了CITED1基因沉默的K1细胞株。沉默CITED1基因表达后,K1细胞的增殖能力明显降低(P <0.01),细胞凋亡率明显升高(P=0.001),G0/G1期细胞所占比例明显增高(P=0.007),G2/M及S期细胞所占比例均明显降低(P值均<0.05),而且12 h和24 h时细胞迁移能力明显减弱(P值均<0.01)。结论:沉默CITED1基因表达可以抑制甲状腺乳头状癌K1细胞的增殖和迁移,促进肿瘤细胞凋亡。

Objective： To investigate the effects of Cbp/p300 interacting transactivator with Glu/Asp rich carboxy-terminal domain 1 （CITED1） gene silencing on the proliferation, apoptosis and migration of papillary thyroid carcinoma K1 cells. Metbods： The recombinant lentivirus carrying CITEDI-shRNA or the negative control （NC）-shRNA was successfully infected into thyroid cancer K1 cells. The silencing efficiency of CITED1 gene was detected by real-time fluorescent quantitative PCR and Western blotting, respectively. The cell proliferation was detected by CCK-8 method, the cell cycle distribution and apoptosis were detected by FCM, and the cell migration ability was detected by scratch wound healing assay. Results： After the recombinant lentivirus carrying CITEDI-shRNA was successfully infected into papillary thyroid carcinoma K1 cells, the expression levels of CITED1 mRNA and protein were significantly decreased （both P 〈 0.01）, which suggested that the K1 cells with CITED1 gene silencing were successfully obtained. After silencing CITED1 gene expression, the cell proliferation was significantly inhibited （P 〈 0.01）, the apoptosis rate was significantly increased （P = 0.001）, the proportion of G0/Gl-phase cells was significanlty increased （P = 0.007）, the proportion of G2/M- and S-phase cells was significanlty decreased （both P 〈 0.05）, and the cell migration abilities at 12 h and 24 h were significantly decreased （both P 〈 0.01）. Conclusion： Silencing CITED1 gene expression can inhibit the proliferation and migration of papillary thyroid carcinoma K1 cells, and promote the apoptosis of tumor cells.