CCNB1基因通过调控PI3K/Akt信号通路对口腔鳞癌细胞增殖、侵袭和迁移的影响

目的 探讨细胞周期蛋白B1(CCNB1)基因对口腔鳞癌细胞增殖、侵袭和迁移的影响及作用机制。方法 实时荧光定量PCR(qRT-PCR)检测人正常口腔上皮细胞系HOK和人口腔鳞癌细胞系SCC-15、SCC-4、Cal-27中CCNB1基因的表达量,将CCNB1 siRNA(si-CCNB1)和阴性对照(si-NC)转染至SCC-15细胞,同时设置空白对照(Control),qRT-PCR和Western blot检测各组SCC-15细胞中CCNB1的表达,MTT实验、Transwell实验和划痕实验分别检测沉默CCNB1对SCC-15细胞增殖、侵袭和迁移能力的影响。Western blot检测细胞中MMP-2、MMP-9、PI3K、Akt和p-Akt蛋白的表达。结果 CCNB1在口腔鳞癌细胞系中的表达显著高于正常口腔上皮细胞(P＜0.05),si-NC组SCC-15细胞中CCNB1 mRNA和蛋白的表达与Control组无统计学差异(P＞0.05);si-CCNB1组中CCNB1 mRNA和蛋白的表达明显低于si-NC组和Control组(P＜0.05),si-NC组SCC-15细胞增殖、侵袭和迁移能力及MMP-2、MMP-9、PI3K、Akt、p-Akt蛋白表达与Control组无统计学差异(P＞0.05);si-CCNB1组细胞增殖、侵袭和迁移能力及MMP-2、MMP-9、PI3K、p-Akt蛋白表达均明显低于si-NC组和Control组(P＜0.05),两组间Akt蛋白表达无统计学差异(P＞0.05)。结论 CCNB1在口腔鳞癌细胞系中呈高表达,沉默CCNB1基因能够抑制人口腔鳞癌SCC-15细胞增殖、侵袭和迁移,其作用机制可能与抑制PI3K/Akt信号通路的激活有关。

Effect of CCNB1 gene on proliferation,invasion and migration of oral squamous cell carcinoma cells by regulating PI3K/Akt signaling pathway

Objective The aim of this study was to investigate the effect and mechanism of cyclin B1(CCNB1)gene on the proliferation,invasion and migration of oral squamous cell carcinoma.Methods Quantitative real-time fluorescent PCR(qRT-PCR)was used to detect the expression of CCNB1 gene in human normal oral epithelial HOK cell line and human oral squamous cell carcinoma SCC-15,SCC-4 and Gal-27 cell lines.CCNB1 siRNA(si-CCNB1)and negative control(si-NC)were transfected into SCC-15 cells.At the same time,blank control(Control)was set.qRT-PCR and Western blot were used to detect the expression of CCNB1 in each group of SCC-15 cells.MTT,Transwell and Scratch assays were used to detect the effects of silencing CCNB1 on the proliferation,invasion and migration of SCC-15 cells.Western blot was used to detect the expression of MMP-2,MMP-9,PI3K,Akt and p-Akt in SCC-15 cells.Results The expression of CCNB1 in oral squamous carcinoma cells was significantly higher than that in normal oral epithelial cells(P<0.05).The expression of CCNB1 mRNA and protein in SCC-15 cells in the si-NC group was not different from that of the control group(P>0.05),The expression of CCNB1 mRNA and protein in the si-CCNB1 group was significantly lower than that in the si-NC group and control group(P<0.05),The proliferation,invasion and migration ability of SCC-15 cells in the si-NC group and the expression of MMP-2,MMP-9,PI3K,Akt and p-Akt in the si-NC group were not significantly different from those in the control group(P>0.05).The cell proliferation,invasive and migratory abilities,and the expressions of MMP-2,MMP-9,PI3K and p-Akt in the si-CCNB1 group were significantly lower than those in the si-NC group and control group(P<0.05).There was no difference in the expression of Akt protein between the si-CCNB1 and si-NC groups(P>0.05).Conclusion CCNB1 is highly expressed in oral squamous carcinoma cells.Silencing CCNB1 gene can inhibit the proliferation,invasion and migration of human oral squamous carcinoma SCC-15 cells.The mechanism may be related to the inhibitory activation of PI3K/Akt signaling pathway.