KLF4转染对口腔鳞癌细胞侵袭能力的影响

KLF4 transduction promotes oral squamous cell carcinoma cells invasion

目的观察KLF4转染对人口腔鳞癌细胞系SCC15侵袭能力的影响。方法培养人口腔鳞癌细胞系SCC15,构建pReceiver-Lv105/KLF4慢病毒表达载体,包装病毒颗粒,感染SCC15后,puromycin筛选KLF4稳定表达的细胞系。荧光显微镜和免疫荧光细胞化学鉴定病毒感染成功。然后通过划痕实验检测KLF4转导对细胞体外迁移能力的影响。Transwell细胞侵袭实验检测KLF4转导对细胞侵袭能力的影响。实时荧光定量PCR检测与细胞侵袭和迁移密切相关的因子MMP-9和Twist1的表达。结果与对照载体转导组SCC15/Lv105细胞相比,SCC15/KLF4细胞中KLF4的表达明显增强。SCC15/KLF4细胞的相对迁移率显著高于SCC15/Lv105细胞(P<0.01)。SCC15/KLF4组的侵袭穿膜细胞数明显多于SCC15/Lv105组(P<0.05)。另外,SCC15/KLF4细胞中MMP-9和Twist1 RNA的表达也高于对照组SCC15/Lv105细胞,其中MMP-9 RNA的表达升高更为显著(P<0.01)。结论KLF4转导SCC15细胞后,可能通过增强MMP-9和Twist1的表达促进细胞的迁移和侵袭能力。

Objective To investigate the effect of KLF4 transduction on invasion of oral squamous cell carcinoma （OSCC） cell line SCC15. Methods OSCC cell line SCC15 was cultured in DMEM, F12 medium with 10% FBS. Lentiviruses carrying KLF4 gene were constructed and transducted into SCC15. Puromycin was used to select the stable cell line expressing KLF4 gene. Fluorescence microscope and immnnoeytochemistry were used to identify the success of transduetion. Cellular migration was assessed by wound-healing assays. The invasion of infected cells was examined by cell invasion assay. The expressions of MMP-9 and Twistl that are associated with cellular invasion and migration were detected by RT-PCR. Results The expression of KLF4 in SCC15/KLF4 cells was significantly elevated compared with SCC15/ Lvl05 cells. The relative migration rate of SCC15/KLF4 cells was significantly increased than that of SCC15/Lv105 cells （ P 〈 0.01 ）. The numbers of cells that crossed the matrigel and membrane in KLF4 infected cells were much more than control （P 〈 0. 05 ）. In addition, the expressions of MMP-9 and Twist1 RNA in SCC15/KLF4 cells were also higher than that of SCC15/Lv105 ceils, and the expression of MMP-9 RNA in SCC15/KLF4 ceils was significantly higher than that of SCC15/Lv105 cells（P 〈0. 01）. Conclusion SCC15 cells transduction with KLF4 promote the migration and invasion of SCC15 cells in vitro by enhancing the expressions of MMP-9 and Twist1.