LINC00665通过靶向miR-708-5p/DKK3抑制食管癌细胞的增殖及侵袭  

LINC00665 inhibits proliferation and invasion of esophageal cancer cells via targeting miR-708-5p/DKK3 axis

作　　者：吕翠婷[1] 尹情 韩俊淑 董稚明[2] 郭炜[2] 沈素朋[2] 梁佳[2] 郭艳丽[2] LÜCui-ting;YIN Qing;HAN Jun-shu;DONG Zhi-ming;GUO Wei;SHEN Su-peng;LIANG Jia;GUO Yan-li(Department of Reproductive Medicine,the Fourth Hospital of Hebei Medical University,Shijiazhuang 050011,China;Hebei Cancer Institute,the Fourth Hospital of Hebei Medical University,Shijiazhuang 050011,China)

机构地区：[1]河北医科大学第四医院生殖科,石家庄050011 [2]河北医科大学第四医院肿瘤研究所,石家庄050011

出　　处：《临床与实验病理学杂志》2022年第4期410-415,共6页Chinese Journal of Clinical and Experimental Pathology

基　　金：河北省自然科学基金(H2020206368);河北省人才工程培养资助项目(201901035)。

摘　　要：目的探讨LINC00665在食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)组织及细胞中的表达,并探讨其异常表达对ESCC细胞增殖、侵袭能力的影响及可能的分子机制。方法采用qRT-PCR法检测LINC00665及miR-708-5p的表达情况;采用MTS、Transwell侵袭实验检测LINC00665异常表达对ESCC细胞增殖及侵袭能力的影响。采用双荧光素酶报告基因检测LINC00665与miR-708-5p的相互作用及miR-708-5p对靶基因的调控作用。结果ESCC组织中LINC00665表达量(3.834±2.016)明显低于癌旁正常组织(7.973±2.752)(P<0.01)。LINC00665表达与ESCC淋巴结转移、浸润深度及TNM分期密切相关(P均<0.05)。LINC00665过表达明显抑制TE13细胞的增殖及侵袭能力(P均<0.05)。LINC00665过表达降低TE13细胞中miR-708-5p的表达及报告基因的荧光素酶活性(P均<0.05);miR-708-5p mimics可降低DKK3基因表达及含DKK33’UTR区结合位点片段的报告基因质粒的荧光素酶活性(P均<0.05);同时,miR-708-5p过表达可抵消TE13细胞中由LINC00665过表达诱导的DKK3基因表达上调和荧光素酶活性的上调(P均<0.05)。结论LINC00665可通过竞争结合miR-708-5p靶向调控DKK3基因表达,进而抑制ESCC细胞的增殖及侵袭能力。LINC00665有望成为ESCC患者分子靶向治疗的潜在靶点及预后评估的新型分子标志物。Purpose To investigate the expression of LINC00665 in esophageal squamous cell carcinoma(ESCC)samples and cell lines,as well as the effect of LINC00665 on the proliferation,invasion and its possible molecular mechanism.Methods The expression levels of LINC00665 and miR-708-5p were detected by qRT-PCR method.MTS and Transwell assay were performed to assess the effect of LINC00665 on proliferation and invasion of esophageal cancer cells.The interaction between LINC00665 and miR-708-5p,as well as the miR-708-5p and DKK3 gene were detected by double fluorescent reporter gene.Results The expression level of LINC00665 in ESCC was 3.834±2.016,which was significantly lower than that in adjacent normal tissues(7.973±2.752,P<0.01).Low expression of LINC00665 was correlated with lymph node metastasis(P<0.01),depth of invasion(P<0.01),TNM stage(P<0.05).Overexpression of LINC00665 significantly reduced proliferation(P<0.05)and invasion ability(P<0.05)in TE13 cells.Overexpression of LINC00665 noticeably reduced miR-708-5p expression level(P<0.05)and the luciferase activity(P<0.05)in TE13 cells.In addition,miR-708-5p mimics lowered the expression level(P<0.05)and luciferase activity of reporter plasmid containing DKK33’UTR binding site fragment(P<0.05).Meanwhile,overexpression of miR-708-5p could counteract the corresponding upregulated expression(P<0.05)and luciferase activity of DKK3 induced by LINC00665 overexpression in TE13 cells(P<0.05).Conclusion LINC00665 could regulate the expression of DKK3 gene via competitive binding miR-708-5p further to promote the proliferation and invasion of ESCC cells.LINC00665 is expected to provide potential targets for molecular therapy and the novel molecular markers for the prognosis evaluation of ESCC patients.

关 键 词：食管肿瘤 鳞状细胞癌 LINC00665 miR-708-5p DKK3基因 

分 类 号：R735.1[医药卫生—肿瘤]