摘要
目的探究CD168对口腔鳞状细胞癌增殖、侵袭的作用机制。方法比较人正常口腔角质细胞系HOK与不同口腔鳞癌细胞株间CD168表达差异,选择HN13细胞株作为研究对象,感染慢病毒shRNA载体后,实时荧光定量PCR(RT-qPCR)和Western印迹法检测CD168基因和蛋白表达检测干预效果,CCK-8法检测细胞增殖情况,流式细胞仪检测细胞凋亡率,Transwell试验检测细胞侵袭情况,Western印迹检测细胞增殖、凋亡、侵袭以及CXCL12-CXCR4/CXCR7信号轴相关蛋白表达。结果选择HN13细胞和CD168-shRNA2慢病毒载体进行后续实验(P<0.05);沉默CD168可使HN13细胞增殖、侵袭数量减少,细胞凋亡率升高(P<0.05),VEGF、PCNA、MMP-2、MMP-9、CXCL12、CXCR4、CXCR7蛋白表达量降低(P<0.05),Bax/Bcl-2比值升高(P<0.05),shRNA-NC组变化无统计学意义(P>0.05)。结论靶向沉默CD168可抑制口腔鳞状癌细胞HN13增殖、侵袭,促进其凋亡,可能与CXCL12-CXCR4/CXCR7信号轴有关。
Objective To explore the effect of CD168 on the proliferation and invasion of oral squamous cell carcinoma and its underlying mechanism.Methods The expression level of CD168 in the human normal oral keratinocyte cell line HOK and several oral squamous cell carcinoma cell lines was compared,and HN13 cell line was selected for the follow-up researches.Gene and protein expression levels of CD168 in cells infected with CD168 shRNA lentivirus were detected by using RT-qPCR and Western blotting.CCK-8 method was applied for the assay of the cell proliferation,the flow cytometry was used for the assay of cell apoptosis and the transwell assay was adopted to detect the cell invasion.Western blotting was performed to detect the expression levels of proteins related to cell proliferation,apoptosis,invasion and CXCL12-CXCR4/CXCR7 signaling axis.Results The HN13 control cell line and the CD168-shRNA2 cell line were used for further studies(P<0.05).Silencing CD168 could suppress the proliferation and invasion of HN13 cells and increase its apoptosis rate(P<0.05).The protein expression levels of VEGF,PCNA,MMP-2,MMP-9,CXCL12,CXCR4 and CXCR7 were decreased in the shCH168 cells(P<0.05),while the Bax/Bcl2 ratio was increased(P<0.05),with no statistical changes observed between the shRNA-NC group and the control group(P>0.05).Conclusion Silencing of CD168 can inhibit the proliferation and invasion of oral squamous cell carcinoma cell line HN13,and promote their apoptosis,the mechanism may be related to the CXCL12-CXCR4/CXCR7 signaling axis.
作者
刘昕
李天客
杜宁
路月亭
刘学聪
LIU Xin;LI Tianke;DU Ning;LU Yueting;LIU Xuecong(Department of Stomatology,Fourth Hospital of Hebei Medical University,Shijiazhuang,050011,China;Department of Stomatology,Children’s Hospital of Hebei Province,Shijiazhuang,050031,China)
出处
《医学分子生物学杂志》
CAS
2022年第6期457-463,共7页
Journal of Medical Molecular Biology
基金
河北省卫健委重点科技研究计划(No.20180526)。
