分泌磷酸蛋白1对肺腺癌A549细胞生物学特性的影响  

Effects of SPP1 on biological characteristics of lung adenocarcinoma A549 cells

作　　者：栾艳超[1] 梁超[1] 韩青松[1] 刘佳坤[1] 杨立伟[2] 李志峰 LUAN Yanchao;LIANG Chao;HAN Qingsong;LIU Jiakun;YANG Liwei;LI Zhifeng(Department of Thoracic Surgery,Hebei Chest Hospital,Shijiazhuang 050001,China;Department of Thoracic Surgery,the Fourth Hospital of Hebei Medical University)

机构地区：[1]河北省胸科医院胸外科,050001 [2]河北医科大学第四医院胸外科

出　　处：《天津医药》2022年第5期449-455,共7页Tianjin Medical Journal

基　　金：河北省卫生健康委科研基金项目(20191004)。

摘　　要：目的探讨分泌磷酸蛋白1(SPP1)在肺腺癌(LUAD)中的表达情况及对LUAD细胞生物学功能的影响。方法通过UALCAN、GEPIA数据库分析SPP1 mRNA在LUAD中的表达情况,GEPIA、K-M数据库分析SPP1 mRNA表达与预后的关系;通过实时荧光定量PCR(qPCR)和Western blot检测SPP1在LUAD组织及细胞系中表达情况。构建SPP1干扰序列并将其转染入A549细胞中,设为Blank组(不转染)、NC组(转染NC序列)、si SPP1组(转染si SPP1序列)。Western blot检测细胞中SPP1沉默效果;CCK-8、EdU、细胞克隆形成实验检测si SPP1对LUAD细胞增殖能力的影响;TUNEL实验检测si SPP1对LUAD细胞凋亡的影响;Transwell和划痕实验检测si SPP1对LUAD细胞侵袭和迁移能力的影响。结果生物信息数据库分析显示,SPP1 mRNA在LUAD中高表达(P<0.05),且SPP1 mRNA高表达者累积生存率较低(P<0.05);LUAD组织及A549细胞中SPP1均高表达(P<0.05);沉默SPP1表达能够抑制A549细胞的增殖、侵袭和迁移能力,诱导细胞凋亡(P<0.05)。结论SPP1可作为肺腺癌诊治的潜在靶点,其可能通过调控细胞增殖、迁移及侵袭等特性影响肺腺癌的发生、发展。Objective(LUAD)and its effect on the biological function of LUAD cells.Methods analyzed by UALCAN and GEPIA databases.The relationship between SPP1 mRNA expression and prognosis was analyzed by GEPIA and K-M database.The expression levels of SPP1 in LUAD tissues and cell lines was detected by real-time fluorescence quantitative PCR(qPCR)and Western blot assay.The interfering sequence targeting SPP1 was constructed and transfected into A549 cells,and cells were divided into the blank group(no transfection),the NC group(transfection with SPP1 negative control sequence)and the si SPP1 group(transfection with siRNA sequence targeting SPP1).Western blot assay was used to detect the silencing effect of SPP1 in cells.The effects of si SPP1 on the proliferation of LUAD cells were detected by CCK-8,EDU and plate clonal formation experiments.TUNEL apoptosis assay was used to detect the effect of si SPP1 on apoptosis of LUAD cells.The effects of si SPP1 on the invasion and migration of LUAD cells were detected by Transwell and scratch assay.ResultsBioinformation database analysis showed that SPP1 mRNA was highly expressed in LUAD(P<0.05).The high expression of SPP1 mRNA was negatively correlated with the overall survival(OS)of patients with LUAD(P<0.05).SPP1 was highly expressed in LUAD and A549 cells(P<0.05).Silencing SPP1 could inhibit the proliferation,invasion and migration of A549 cells and induce apoptosis(P<0.05).Conclusion SPP1can be a potential target in the diagnosis and treatment of lung adenocarcinoma,which may affect the occurrence and development of lung adenocarcinoma by regulating cell proliferation,migration and invasion.

关 键 词：肺肿瘤 腺癌 细胞增殖 细胞凋亡 细胞运动 分泌磷酸蛋白1 

分 类 号：R734.2[医药卫生—肿瘤]