c-Jun通过调控RNF144A-AS1转录影响食管鳞状细胞癌恶性生物学行为OA
c-Jun regulated the malignant biological behaviors of esophageal squamous cell carcinoma through transcriptional activation of RNF144A-AS1
目的 探讨c-Jun通过调控RNF144A-AS1转录影响食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)细胞的增殖、迁移以及侵袭能力.方法 常规培养人食管上皮细胞HEEpiC和ESCC细胞Eca109、Kyse170、Kyse150、TE1,用转染试剂将敲减序列和过表达质粒转染ESCC细胞.qRT-PCR法检测c-Jun mRNA和RNF144A-AS1在ESCC组织、不同ESCC细胞系以及各组Eca109细胞中的表达,Western blot法检测c-Jun的敲减效率,应用MTS实验检测各组Eca109细胞的增殖能力,划痕愈合和Transwell小室实验检测各组Eca109细胞的迁移和侵袭能力,利用生物信息学预测c-Jun与RNF144A-AS1启动子区的结合位点,染色质免疫沉淀实验和双荧光素酶报告基因实验验证c-Jun与RNF144A-AS1启动子区之间的结合关系.结果 qRT-PCR检测显示RNF144A-AS1和c-Jun在ESCC组织[1.10(0.44,2.39);1.02(0.35,4.39)]中的表达高于癌旁正常组织[0.40(0.15,1.76);0.51(0.35,1.61),P均<0.05].qRT-PCR检测显示RNF144A-AS1 和c-Jun在ESCC细胞系Eca109(15.54±1.74;15.14±1.52)、Kyse170(10.92±0.70;9.62±0.86)、Kyse150(8.91±0.18;7.68±0.46)、TE1(4.75±0.54;6.11±0.67)中的表达均高于HEEpiC(1.15±0.13;1.02±0.05,P均<0.01).GEPIA数据分析显示,在食管癌中RNF144A-AS1和c-Jun表达呈正相关(r=0.24,P=0.000 89);在ESCC组织中,RNF144A-AS1和c-Jun表达亦呈正相关(r=0.456,P<0.01);c-Jun蛋白表达水平与pTNM分期有相关性(P<0.05);敲减RNF144A-AS1后可抑制Eca109细胞的增殖、迁移、侵袭能力(P<0.05);敲减或过表达c-Jun后RNF144A-AS1表达量显著降低或升高(P<0.01);c-Jun与RNF144A-AS1启动子区直接结合并转录激活其表达(P<0.05);过表达RNF144A-AS1可部分逆转敲减c-Jun对Eca109细胞恶性生物学行为的影响(P<0.05).结论 c-Jun通过转录激活RNF144A-AS1参与ESCC细胞的恶性生物学行为.
Objective This study aimed to investigate the role of c-Jun in regulating the transcription of RNF144A-AS1 and its impact on the proliferation,migration and invasion of esophageal squamous cell carcinoma(ESCC)cells.Methods Human esophageal epithelial cell(HEEpiC)and ESCC cell lines(Eca109,Kyse170,Kyse150 and TE1)were cultured under standard conditions.Knockdown sequences and overexpression plasmids were transfected into ESCC cells using transfection reagents.The expression of c-Jun mRNA and RNF144A-AS1 in ESCC tissues,different cell lines,and transfected Eca109 cells was detected by qRT-PCR.Western blotting was performed to evaluate the knockdown efficiency of c-Jun.Cell proliferation,migration,and invasion were examined using MTS,wound healing,and Transwell assays.Bioinformatic analysis was applied to predict the binding sites of c-Jun within the RNF144A-AS1 promoter region,which were further validated by ChIP and dual-luciferase reporter assays.Results qRT-PCR showed that RNF144A-AS1 and c-Jun expression levels in ESCC tissues[1.10(0.44,2.39);1.02(0.35,4.39)]were higher than in adjacent normal tissues[0.40(0.15,1.76);0.51(0.35,1.61),all P<0.05].Similarly,their expression levels in ESCC cell lines Eca109(15.54±1.74;15.14±1.52),Kyse170(10.92±0.70;9.62±0.86),Kyse150(8.91±0.18;7.68±0.46),and TE1(4.75±0.54;6.11±0.67)were markedly el-evated compared with HEEpiC cells(1.15±0.13;1.02±0.05,all P<0.01).GEPIA dataset analysis revealed a positive correlation between RNF144A-AS1 and c-Jun expression in esophageal cancer tissues(r=0.24,P=0.000 89),which was also observed in ESCC tissues(r=0.456,P<0.01).c-Jun protein expression was significantly correlated with pTNM stage(P<0.05).Knockdown of RNF144A-AS1 suppressed proliferation,migration,and invasion of Eca109 cells(P<0.05).siRNA-mediated c-Jun knockdown reduced RNF144A-AS1 expression,whereas c-Jun over-expression increased it(P<0.01).c-Jun directly bound to the RNF144A-AS1 promoter and transcriptionally acti-vated its expression(P<0.05).Overexpression of RNF144A-AS1 partially reversed the inhibitory effects of c-Jun knockdown on the malignant behaviors of Eca109 cells(P<0.05).Conclusion c-Jun promotes the malignant bio-logical behavior of ESCC cells by transcriptionally activating RNF144A-AS1.
冯博;曹家瑞;许彦超;陈伟霞;马纯政
河南省中医院肿瘤科,郑州 450003河南中医药大学研究生院,郑州 450046河南省中医院肿瘤科,郑州 450003河南省中医院肿瘤科,郑州 450003河南省中医院肿瘤科,郑州 450003
医药卫生
食管肿瘤鳞状细胞癌长链非编码RNA转录调控c-JunRNF144A-AS1
esophageal neoplasmssquamous cell carcinomalong non-coding RNAtranscriptional regulationc-JunRNF144A-AS1
《临床与实验病理学杂志》 2026 (1)
46-55,10
河南省重点研发与推广专项(科技攻关)项目(232102310126)、河南省中医院(河南中医药大学第二附属医院)博士科研基金(2021BSJJ04) Henan Provincial Key R&D and Promotion Special(Scientific and Technological Research)Project(232102310126)Henan Province Hospital of Traditional Chinese Medicine(the Second Affiliated Hospital of Henan University of Chinese Medicine)Doctoral Research Fund(2021BSJJ04)
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