FAK基因沉默促进舌癌细胞失巢凋亡及抑制转移的实验研究

Focal adhesion kinase gene silencing can promote anoikis and suppresses metastasis of human tongue cancer cell line

目的:研究黏着斑激酶(FAK)基因沉默是否可促进舌癌细胞系Tca8113失巢凋亡,进而逆转其失巢凋亡抗性,抑制舌癌细胞侵袭转移。方法:以舌癌细胞株Tca8113为研究对象,首先利用RNA干扰方法抑制FAK基因表达。采用poly-HEMA诱导悬浮培养,进而采用流式细胞仪观察体外培养过程中,FAK基因沉默对舌癌细胞株Tca8113失巢凋亡抗性的影响。最后以划痕实验、Transwell小室侵袭模型研究舌癌细胞迁移及侵袭能力。结果 :在体外培养过程中,Tca8113细胞具有很强的失巢凋亡抗性。针对FAK的siRNA可显著下调FAK基因的表达。当FAK基因下调后,Tca8113细胞失巢凋亡抗性显著下降(P<0.01),并且细胞的侵袭及迁移能力明显下降(P<0.01)。结论:FAK基因沉默可促进失巢凋亡,逆转舌癌细胞失巢凋亡抗性,进而抑制其转移,是一个潜在的抗舌癌转移分子治疗的靶点。

Objective：To investigate the roles of FAK in the process of invasion, migration and anoikis resistance in the human tongue cancer cell line, Tca8113. Methods： Expression of FAK was suppressed by RNA interference method with pSilencer-FAK vector in Tca8113 cells. Anoikis was induced in Tca8113 by the use of polyHEMA culture, and anoikis was detected by flow cytometry. The velocity of cell migration was measured by wound-repair experiment, and the invasion of Tca8113 was detected by Invasion chamber assay. Results：Tca8113 cells showed remarkable anoikis resis-tance, which was associated with higher levels of FAK expression. FAK gene silencing could promote anoikis remarkably in Tca8113 （P〈0.01）. After FAK gene silencing, the velocity of cells migration and the invasion of cells were suppressed sig-nificantly in Tca8113 cells. Conclusion：RNAi-mediated FAK reduction decreased tongue cancer cell migration, invasion by reversing acquired anoikis resistance. Moreover, RNAi targeting FAK could serve as a potential therapeutic for the treat-ment of tongue cancer.