环状RNA circPRDM5在食管鳞状细胞癌中的表达和功能研究
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摘要
目的探索环状RNA circPRDM5在食管鳞状细胞癌(ESCC)细胞和组织中的表达水平,及其对ESCC细胞功能的影响。方法 qRT?PCR检测circPRDM5在正常食管上皮细胞(HEEC)、ESCC细胞系、和44例ESCC组织及对应的癌旁正常组织中的相对表达水平;Transwell实验和CCK8实验分别检测siRNA沉默circPRDM5后,对细胞迁移侵袭和增殖功能的影响;构建裸鼠成瘤模型,检测circPRDM5对ESCC细胞在动物体内成瘤能力的影响。结果与HEEC相比,circPRDM5在ESCC细胞系KYSE150、ECA109中高表达(P<0.05),且在ESCC组织中表达(2.90±0.18)较癌旁正常组织(2.01±0.15)表达高(P<0.001)。Transwell实验和CCK8实验证明,沉默circPRDM5能明显抑制ESCC细胞的迁移侵袭和增殖能力(P<0.05);裸鼠成瘤实验结果表明,沉默circPRDM5后,ESCC细胞的成瘤体积和重量较对照组明显下降(P<0.01)。结论 circPRDM5在ESCC的细胞和组织中高表达。在ESCC细胞中沉默circPRDM5能降低ESCC细胞的增殖、迁移侵袭、和在动物体内的成瘤能力。提示circPRDM5可能成为ESCC治疗的一个新靶点。
Objective To investigate the expression level of circular RNA circPRDM5 in esophageal squamous cell carcinoma(ESCC)cells and tissues,and its effect on ESCC cell function. Methods qRT-PCR was used to detect the relative expression of circPRDM5 in normal esophageal epithelial cells(HEEC),ESCC cell line,and 44 ESCC tissues and corresponding adjacent normal tissues;Transwell and CCK8 assays were performed to detect the effects on cell migration,invasion and proliferation after siRNA silencing circPRDM5;a nude mouse model was established to examine the effect of circPRDM5 on the tumorigenic ability of ESCC cells in animals. Results Compared with HEEC,circPRDM5 was highly expressed in ESCC cell lines KYSE150 and ECA109(P<0.05),and the expression in ESCC tissues(2.90±0.18)was higher than that in adjacent normal tissues(2.01±0.15)(P<0.001);Transwell assay and CCK8 assay demonstrated that silencing of circPRDM5 significantly inhibited the migration,invasion and proliferation of ESCC cells(P<0.05). The results of tumor formation in nude mice showed that the volume and weight of ESCC cells after silencing circPRDM5 were significantly lower than those of the control group(P<0.01). Conclusion circPRDM5 is highly expressed in the cells and tissues of ESCC. Silencing circPRDM5 in ESCC cells reduced the proliferation,migration and invasion of ESCC cells,and the vivo tumorigenesis abilities. It is suggested that circPRDM5 may become a new target for ESCC treatment.
Objective To investigate the expression level of circular RNA circPRDM5 in esophageal squamous cell carcinoma(ESCC)cells and tissues,and its effect on ESCC cell function. Methods qRT-PCR was used to detect the relative expression of circPRDM5 in normal esophageal epithelial cells(HEEC),ESCC cell line,and 44 ESCC tissues and corresponding adjacent normal tissues;Transwell and CCK8 assays were performed to detect the effects on cell migration,invasion and proliferation after siRNA silencing circPRDM5;a nude mouse model was established to examine the effect of circPRDM5 on the tumorigenic ability of ESCC cells in animals. Results Compared with HEEC,circPRDM5 was highly expressed in ESCC cell lines KYSE150 and ECA109(P<0.05),and the expression in ESCC tissues(2.90±0.18)was higher than that in adjacent normal tissues(2.01±0.15)(P<0.001);Transwell assay and CCK8 assay demonstrated that silencing of circPRDM5 significantly inhibited the migration,invasion and proliferation of ESCC cells(P<0.05). The results of tumor formation in nude mice showed that the volume and weight of ESCC cells after silencing circPRDM5 were significantly lower than those of the control group(P<0.01). Conclusion circPRDM5 is highly expressed in the cells and tissues of ESCC. Silencing circPRDM5 in ESCC cells reduced the proliferation,migration and invasion of ESCC cells,and the vivo tumorigenesis abilities. It is suggested that circPRDM5 may become a new target for ESCC treatment.
引文
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[2]Torre LA,Bray F,Siegel RL,et al.Global cancer statistics,2012[J].CA Cancer J Clin,2015,65(2):87-108.
[3]Sjoquist KM,Burmeister BH,Smithers BM,et al.Survival after neoadjuvant chemotherapy or chemoradiotherapy for resectable oesophageal carcinoma:an updated meta-analysis[J].Lancet Oncol,2011,12(7):681-692.
[4]Fakhrian K,Ordu AD,Lordick F,et al.Long-term outcomes of trimodality treatment for squamous cell carcinoma of the esophagus with cisplatin and/or 5-FU:more than 20 years'experience at a single institution[J].Strahlenther Onkol,2014.190(12):1133-1140.
[5]Chen LL,Yang L.Regulation of circRNA biogenesis[J].RNABiology,2015,12(4):381-388.
[6]Piwecka M,Glazar P,Hernandez-Miranda LR,et al.Loss of a mammalian circular RNA locus causes miRNA deregulation and affects brain function[J].Science,2017,357(6357).
[7]Li Y,Zheng F,Xiao X,et al.CircHIPK3 sponges miR-558 to suppress heparanase expression in bladder cancer cells[J].EMBO Rep,2017,18(9):1646-1659.
[8]Hall IF,Climent M,Quintavalle M,et al.Circ_Lrp6,a circular RNA enriched in vascular smooth muscle cells,acts as a sponge regulating miRNA-145 function[J].Circ Res,2019,124(4):498-510.
[9]Jeck WR,Sharpless NE.Sharpless,Detecting and characterizing circular RNAs[J].Nat Biotechnol,2014,32(5):453-461.
[10]Vicens Q,Westhof E.Biogenesis of Circular RNAs[J].Cell,2014,159(1):13-14.
[11]Bose R,Ain R.Regulation of Transcription by Circular RNAs[J].Adv Exp Med Biol,2018,1087:81-94.
[12]Zhang H,Wang X,Huang H,et al.Hsa_circ_0067997 promotes the progression of gastric cancer by inhibition of miR-515-5p and activation of X chromosome-linked inhibitor of apoptosis(XIAP)[J].Artif Cells Nanomed Biotechnol,2019.47(1):308-318.
[13]Zheng H,Chen T,Li C,et al.A circular RNA hsa_circ_0079929 inhibits tumor growth in hepatocellular carcinoma[J].Cancer Management and Research,2019,11:443-454.
[14]Qiu BQ,Zhang PF,Xiong D,et al.CircRNA fibroblast growth factor receptor 3 promotes tumor progression in non-small cell lung cancer by regulating Galectin-1-AKT/ERK1/2 signaling[J].J Cell Physiol,2019,234(7):11256-11264.
[15]Akhter R.Circular RNA and Alzheimer′s Disease[J].Adv Exp Med Biol,2018,1087:239-243.
[16]Altesha MA,Ni T,Khan A,et al.Circular RNA in cardiovascular disease[J].J Cell Physiol,2019,234(5):5588-5600.
[17]Jiang C,Xu D,You Z,et al.Dysregulated circRNAs and ceR-NA network in esophageal squamous cell carcinoma[J].Front Biosci(Landmark Ed),2019,24:277-290.
[18]Cao S,Chen G,Yan L,et al.Contribution of dysregulated circRNA_100876 to proliferation and metastasis of esophageal squamous cell carcinoma[J].Onco Targets Ther,2018,11:7385-7394.
[19]Shi N,Shan B,Gu B,et al.Circular RNA circ-PRKCI functions as a competitive endogenous RNA to regulate AKT3 expression by sponging miR-3680-3p in esophageal squamous cell carcinoma[J].J Cell Biochem,2019,120(6):10021-10030.
[20]Di Zazzo E,De Rosa C,Abbondanza C,Moncharmont B.PRDM Proteins:Molecular Mechanisms in Signal Transduction and Transcriptional Regulation[J].Biology(Basel),2013,2(1):107-141.
[21]Meani N,Pezzimenti F,Deflorian G,et al.The tumor suppressor PRDM5 regulates Wnt signaling at early stages of zebrafish development[J].PLoS One,2009,4(1):e4273.
[22]Shu XS,Geng H,Li L,et al.The epigenetic modifier PRDM5functions as a tumor suppressor through modulating WNT/betacatenin signaling and is frequently silenced in multiple tumors[J].PLoS One,2011,6(11):e27346.