ARHI通过Wnt/自噬通路调控口腔鳞癌细胞凋亡的研究  

ARHI induces apoptosis of oral squamous cell carcinoma via Wnt/autophagy-related signaling pathways

作　　者：李建豪 吕少华 李吉辰[1] 朴松林[1] LI Jian-hao;LüShao-hua;LI Ji-chen;PIAO Song-lin(Department of Oral and Maxillofacial Surgery,School of Stomatology,Harbin Medical University,Harbin 150001,China)

机构地区：[1]哈尔滨医科大学口腔医学院口腔颌面外科,黑龙江哈尔滨150001

出　　处：《实用药物与临床》2021年第11期966-973,共8页Practical Pharmacy and Clinical Remedies

基　　金：哈尔滨医科大学附属第一医院科研创新基金(2020M13,2018L004)。

摘　　要：目的探讨Aplasia-Ras同源物成员I(ARHI)在口腔鳞癌(OSCC)中的表达情况,并验证ARHI能否通过Wnt信号通路调控OSCC细胞凋亡。方法通过生物信息学方法整合GEO数据库中OSCC芯片(GSE75540和GSE9844),免疫组化方法检测20例OSCC和癌旁健康组织中ARHI的表达,并用GEPIA和UALCAN数据库评价ARHI在头颈部鳞癌的表达和预后。过表达ARHI后的OSCC细胞(CAL-27以及SCC-115)用CCK-8以及AO/EB方法检测其细胞增殖活性和凋亡情况,通过Western blot以及LC3荧光染色检测过表达ARHI后OSCC细胞(CAL-27、SCC-115)Wnt信号通路相关蛋白以及OSCC细胞自噬的情况。结果GEO口腔鳞癌芯片数据分析结果显示,ARHI在OSCC组织中表达明显下调;生物信息学结果显示,ARHI在头颈部鳞癌中表达降低;20例免疫组化结果显示,ARHI在OSCC组织中的表达降低。OSCC患者的生存曲线分析显示,ARHI的表达水平与OSCC患者的预后相关。过表达ARHI能够很大程度地抑制OSCC细胞(CAL-27、SCC-115)的增殖活性,同时诱导OSCC细胞凋亡。在OSCC细胞中过表达ARHI后,Cleaved-caspase-3、Caspase-3及Bax的表达明显升高,β-catenin蛋白表达降低,GSK-3β表达升高。过表达ARHI后,OSCC细胞(CAL-27、SCC-115)中自噬小体明显增多,且自噬相关蛋白beclin-1表达升高,LC3II/I表达升高。结论ARHI在OSCC组织中低表达,且过表达ARHI能够通过经典Wnt信号通路诱导OSCC细胞发生自噬与凋亡。ARHI可能是OSCC潜在的治疗靶点和预后标志物。Objective To explore the expression of ARHI in OSCC and verify whether ARHI can induces apoptosis of oral squamous cell carcinoma cells via Wnt signaling pathways.Methods OSCC chips(GSE75540 and GSE9844)of GEO database was integrated by bioinformatic method.Immunohistochemical method was used to detect the expression of ARHI in 20 cases of OSCC and normal adjacent tissues.The expression and prognosis of ARHI in head and neck squamous cell carcinoma were evaluated by GEPIA and UALCAN databases.CCK-8 and AO/EB were used to detect the cell proliferation viability and apoptosis in CAL-27 and SCC-115 cells after transfection with ARHI.Western blot and immunofluorescence were used to detect the protein level of Wnt signaling pathway and autophagy in CAL-27 and SCC-115 cells after treated with ARHI.Results Microarray analysis showed that the expression of ARHI was significantly down-regulated in OSCC.Meanwhile,bioinformatic results showed that the expression of ARHI in head and neck squamous cell carcinoma was decreased.The expression of ARHI in OSCC tissues of 20 cases was decreased,which is detected by immunohistochemistry.Survival analysis of OSCC patients also showed that the expression level of ARHI was correlated with the prognosis of patients with OSCC.Overexpression of ARHI significantly inhibited the proliferation activity of Cal-27 and SCC-115 cells and induced apoptosis.Overexpression of ARHI in OSCC cells significantly increased Cleaved caspase-3,Caspase-3,and Bax expression.The expression ofβ-catenin protein was decreased,and the expression of GSK-3βwas increased.After ARHI was overexpressed,the autophagy bodies in Cal-27 and SCC-115 cells were significantly increased,and the expression of Beclin-1 protein and LC3 II/I was increased.Conclusion ARHI is lowly expressed in OSCC tissues,and overexpression of ARHI can induce apoptosis and autophagy of OSCC cells via Wnt-related signaling pathway.ARHI may be a potential therapeutic target and prognostic biomarker in OSCC.

关 键 词：ARHI WNT信号通路 口腔鳞癌 自噬 凋亡 

分 类 号：R739.8[医药卫生—肿瘤]