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目的研究沉默整合素连接激酶(integrin-linked kinase,ILK)基因表达,对人舌鳞癌细胞生长、侵袭和转移能力的影响。方法通过用ILK mRNA的特异性siRNA表达载体和无同源性的对照载体,在脂质体介导下稳定转染人舌鳞癌TCA8113细胞,分为TCA8113组、TCA8113 vector组和TCA8113 siILK组,通过筛选鉴定后,用MTT法、划痕试验和Transwell法分别检测细胞生长、迁移和侵袭能力,用Western blot分析细胞中ILK、p-Akt、p-GSK3β及Snail等的表达。结果沉默ILK基因表达显著抑制了细胞的生长、迁移和侵袭能力,接种的第48、72、96、120小时,TCA8113 siILK组的抑制率分别为17%、29%、25%、42%,迁移能力较TCA8113组和TCA8113 vector组分别下降67%和66%,侵袭能力则较两个对照组下降了67%和68%,p-Akt、p-GSK3β及Snail的表达较TCA8113组和TCA8113 vector组分别降低了45%和53%,57%和61%,74%和73%。结论抑制ILK基因的表达可通过Akt/GSK3β/Snail途径,显著抑制人舌鳞癌TCA8113细胞的生长、迁移和侵袭潜能,可作为治疗人舌鳞癌的靶蛋白。
Objective To investigate the effect of silencing integrin-linked kinase (ILK) gene expression on the growth, invasion and metastasis of human tongue squamous cell carcinoma. METHODS: Human tongue squamous cell carcinoma TCA8113 cells were stably transfected with TCA8113, TCA8113 vector and TCA8113 siILK cells by liposome-mediated siRNA-specific siRNA vector and control vector without homology. After screening, the cell growth, migration and invasion were detected by MTT assay, scratch assay and Transwell assay, respectively. The expression of ILK, p-Akt, p-GSK3β and Snail in cells were detected by Western blot. Results Silencing ILK gene expression significantly inhibited cell growth, migration and invasion. At the 48th, 72th, 96th and 120th hour after inoculation, the inhibitory rates of TCI8113 siILK group were 17%, 29%, 25% and 42%, respectively Compared with the TCA8113 group and the TCA8113 vector group, the ability of invasion was decreased by 67% and 66% respectively, while the invasive ability was decreased by 67% and 68% respectively compared with the two control groups. The expression of p-Akt, p-GSK3β and Snail were significantly lower than those of TCA8113 and TCA8113 vector Decreased by 45% and 53%, 57% and 61%, 74% and 73% respectively. Conclusion Inhibition of ILK gene expression can significantly inhibit the growth, migration and invasion potential of human tongue squamous cell carcinoma TCA8113 cells through the Akt / GSK3β / Snail pathway, which can be used as the target protein in the treatment of human tongue squamous cell carcinoma.