摘要
目的探讨CD109对口腔鳞状细胞癌(OSCC)细胞增殖、迁移和侵袭的影响。方法定量逆转录聚合酶链反应和蛋白质印迹法检测CD109在OSCC细胞中的表达。通过siRNA技术将空载体和CD109-siRNA转染至人舌鳞癌Cal27细胞,并分别作为对照组(siNC)和CD109干扰组(siCD109)。细胞计数试剂盒(CCK-8)和EdU实验检测细胞增殖,流式细胞术检测细胞周期,细胞划痕实验检测细胞迁移能力,Transwell实验检测细胞侵袭能力,基因富集分析检测CD109影响OSCC的潜在机制并通过蛋白质印迹法验证。结果 CD109 mRNA和蛋白在OSCC细胞系中高表达,在口腔上皮细胞中低表达,P<0.001。转染CD109-siRNA后,siCD109组细胞中CD109 mRNA水平为0.27±0.07,siNC组为1.00±0.00,差异有统计学意义,t=18.102,P=0.003。siCD109组CD109蛋白表达水平为0.54±0.10,siNC组为1.41±0.15,差异有统计学意义,t=8.365,P=0.020。siCD109组细胞的生长速度低于siNC组,差异有统计学意义,t=12.571,P<0.001。siCD109组细胞的EdU阳性率[(16.23±3.80)%]低于siNC组[(30.33±5.14)%],差异有统计学意义,t=3.821,P=0.018。相较于siNC组细胞,siCD109组G_(1)期(t=6.151,P=0.004)和G_(2)期(t=5.704,P=0.005)细胞比例增加,S期(t=6.642,P=0.003)细胞比例下降,差异有统计学意义;siCD109组细胞划痕愈合速度[(50.05±4.81)%]低于siNC组[(75.27±5.16)%],差异有统计学意义,t=6.220,P=0.003。siCD109组侵袭细胞数量为(258±45)个,低于siNC组的(455±20)个,差异有统计学意义,t=6.292,P=0.003。siCD109组细胞的PI3K/AKT磷酸化水平较siNC组低,差异有统计学意义,均P<0.05。结论 CD109通过PI3K/AKT信号通路抑制OSCC细胞迁移和侵袭,可能是OSCC潜在的治疗靶点。
Objective To investigate the effects of CD109on proliferation,migration and invasion of oral squamous cell carcinoma(OSCC)cells.Methods Quantitative reverse transcription polymerase chain reaction and Western blot were used to detect the expression of CD109in OSCC cells.The empty vector and CD109-siRNA were transfected into human tongue squamous cell carcinoma Cal27cells by siRNA technology,which were control group(siNC)and CD109interference group(siCD109),respectively.The cell counting kit(CCK-8)and EdU experiments were performed to detect proliferation.The cell cycle was detected by flow cytometry.Wound healing assay was performed to detect cell migration.Transwell assay detected cell invasion.The potential mechanism of CD109affecting OSCC was detected by gene set enrichment analysis and was verified by Western blot.Results CD109mRNA and protein were highly expressed in the OSCC cell line and lowly expressed in oral epithelial cells (P<0.001).After transfection of CD109-siRNA,the level of CD109mRNA in cells in the siCD109group was 0.27±0.07,and that in the siNC group was 1.00±0.00,the difference was statistically significant(t=18.102,P=0.003).The protein expression level of siCD109histone was 0.54±0.10,and that of siNC group was 1.41±0.15,the difference was statistically significant(t=8.365,P=0.020).The growth rate of cells in the siCD109group was lower than that of the siNC group,the difference was statistically significant(t=12.571,P<0.001).The EdU positive rate of cells in the siCD109group[(16.23±3.80)%]was lower than that of the siNC group[(30.33±5.14)%],the difference was statistically significant(t=3.821,P=0.018).Compared with the cells in the siNC group,the proportion of cells in the G_(1)(t=6.151,P=0.004)and G_(2)phases(t=5.704,P=0.005)in the siCD109group increased,and the proportion of cells in the S phase(t=6.642,P=0.003)decreased.The difference was statistically significant.The scratch healing speed of cells in the siCD109group[(50.05±4.81)%]was lower than that of the siNC group[(75.27±5.16)%
作者
侯德强
白宁
魏子程
刘彦魁
程洋
邹建明
HOU De-qiang;BAI Ning;WEI Zi-cheng;LIU Yan-kui;CHENG Yang;ZOU Jian-ming(Affiliated Hospital of Jiangnan University,Wuxi 214000,China;Laboratory of Pathogenic Infection and Immunology,Department of Public Health and Preventive Medicitie,Wuxi Medical College,Jiangnan University,Wuxi 214000,China)
出处
《中华肿瘤防治杂志》
CAS
北大核心
2021年第20期1533-1540,共8页
Chinese Journal of Cancer Prevention and Treatment
基金
无锡市卫健委科研项目(MS201909)
太湖高层次人才(双百中青年医疗卫生拔尖人才)(BJ2020056)。
