LncRNA FGD5-AS1对口腔鳞状细胞癌细胞恶性表型的影响  

Effect of LncRNA FGD5-AS1 on the malignant phenotype of oral squamous cell carcinoma cells

作　　者：杨锋[1] 刘广龙 王艳卿 YANG Feng;LIU Guanglong;WANG Yanqing(Stomatological Department of Tengzhou Central People’s Hospital Affiliated to Jining Medical College,Tengzhou 277599;China.2.Operating Room of Tengzhou Central People’s Hospital,Tengzhou 277599)

机构地区：[1]济宁医学院附属滕州市中心人民医院口腔科,山东滕州277599 [2]滕州市中心人民医院手术室,山东滕州277599

出　　处：《中国比较医学杂志》2023年第11期78-87,共10页Chinese Journal of Comparative Medicine

基　　金：济宁医学院校级科研项目(JY2017FS043)。

摘　　要：目的探讨长链非编码RNA(LncRNA)FGD5-AS1对口腔鳞状细胞癌(OSCC)细胞增殖、凋亡、迁移和侵袭的影响与机制。方法利用在线数据库分析FGD5-AS1在OSCC中的表达。以在滕州市中心人民医院口腔科收集的30例OSCC患者的肿瘤组织、正常组织和体外培养的人口腔黏膜细胞(HOK)和OSCC细胞(SCC-9、HSC-4、SCC-25、CAL-27)为研究对象,采用qRT-PCR法检测FGD5-AS1和miR-129-5p表达。将FGD5-AS1表达最高的CAL-27细胞分成Control组、si-NC组、si-FGD5-AS1组、si-FGD5-AS1+NC inhibitor组和si-FGD5-AS1+miR-129-5p inhibitor组,CCK-8法和克隆形成实验检测细胞增殖能力;流式细胞术检测细胞凋亡水平;划痕愈合实验检测细胞迁移能力;Transwell小室检测细胞侵袭能力;双荧光素酶报告实验验证FGD5-AS1与miR-129-5p的靶向关系;Western blot检测高迁移率族蛋白B1(HMGB1)蛋白表达。构建体内异种移植瘤模型,并分为sh-NC组、sh-FGD5-AS1组、miR-129-5p inhibitor组和sh-FGD5-AS1+miR-129-5p inhibitor组,检测肿瘤体积和肿瘤;qRT-PCR检测移植瘤组织FGD5-AS1、miR-129-5p表达;免疫组化检测移植瘤组织HMGB1、Ki67表达。结果数据库分析显示,OSCC肿瘤组织中FGD5-AS1的表达水平是正常组织的4倍,且FGD5-AS1表达与OSCC患者分级较差相关。与正常组织或人口腔黏膜细胞相比,肿瘤组织和OSCC细胞系中FGD5-AS1表达明显升高,miR-129-5p表达明显降低(P<0.05),选择FGD5-AS1表达水平最高的CAL-27细胞进行转染实验。沉默FGD5-AS1可升高细胞凋亡率,降低细胞活力、划痕愈合率及侵袭细胞数,并增强miR-129-5p表达,下调HMGB1表达(P<0.05)。miR-129-5p是FGD5-AS1的靶基因,抑制miR-129-5p表达可逆转沉默FGD5-AS1对OSCC细胞增殖、凋亡、迁移和侵袭的影响。体内实验显示,沉默FGD5-AS1明显抑制移植瘤生长和HMGB1、Ki67表达(P<0.05),抑制miR-129-5p则相反;抑制miR-129-5p可逆转沉默FGD5-AS1对肿瘤生长和HMGB1、Ki67表达的抑制作用(P<0.05)。结论FGObjectiveTo investigate the effect and mechanism of long non-coding RNA(LncRNA)FGD5-AS1 on the proliferation,apoptosis,migration,and invasion of oral squamous cell carcinoma(OSCC)cells.MethodsFGD5-AS1 expression in OSCC was analyzed using an online database.Tumor and normal tissues of 30 patients with OSCC collected at the Stomatology Department of Tengzhou Central People’s Hospital,and human oral mucosal cell line HOK and OSCC cell lines SCC-9,HSC-4,SCC-25,and CAL-27 cultured in vitro were investigated.qRT-PCR was performed to measure FGD5-AS1 and miR-129-5p expression.CAL-27 cells with the highest FGD5-AS1 expression were divided into Control,si-NC,si-FGD5-AS1,si-FGD5-AS1+NC inhibitor,and si-FGD5-AS1+miR-129-5p inhibitor groups.CCK-8 and colony formation assays were used to assess cell proliferation.Apoptosis was detected by flow cytometry.Cell migration was assessed by wound healing assays.Transwell chambers were used to assess cell invasion.Dual luciferase reporter assays were sued to verify the targeting relationship between FGD5-AS1 and miR-129-5p.Expression of high mobility group protein B1(HMGB1)was detected by Western blot.An in vivo xenograft tumor model was established and divided into sh-NC,sh-FGD5-AS1,miR-129-5p inhibitor,and sh-FGD5-AS1+miR-129-5p inhibitor groups.The tumor volume and tumor were assessed.qRT-PCR was used to measure FGD5-AS1 and miR-129-5p expression in transplanted tumor tissues.HMGB1 and Ki67 expression was detected by immunohistochemistry.ResultsDatabase analysis showed that the expression level of FGD5-AS1 in OSCC tumor tissues was 4 times higher than that in normal tissues.FGD5-AS1 expression was associated with a poor grade in OSCC patients.Compared with normal tissues and human oral mucosal cells,FGD5-AS1 expression in tumor tissues and OSCC cell lines was significantly increased,and miR-129-5p expression was significantly decreased(P<0.05).CAL-27 cells with the highest expression level of FGD5-AS1 were selected for transfection experiments.Silencing FGD5-AS1 increased the a

关 键 词：FGD5-AS1 miR-129-5p 口腔鳞状细胞癌 增殖 凋亡 

分 类 号：R-33[医药卫生]