沉默circROBO1通过下调KRAS抑制鼻咽癌CNE2细胞侵袭与肺转移  

Silencing circROBO1 inhibits the invasion and pulmonary metastasis of nasopharyn-geal carcinoma CNE2 cells by down-regulating KRAS

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作  者:文玲波 孙丕云 罗琴 肖娟 WEN Lingbo;SUN Piyun;LUO Qin;XIAO Juan(The Third Affiliated Hospital of Guangxi University of Chinese Medicine,Liuzhou Traditional Chinese Medicine Hospital,Liuzhou 545001,Guangxi,China;The Second Affiliated Hospital,Hengyang Medical School,University of South China,Hengyang 421001,Hunan,China)

机构地区:[1]广西中医药大学第三附属医院柳州市中医医院,广西柳州545001 [2]南华大学衡阳医学院附属第二医院,湖南衡阳421001

出  处:《中南医学科学杂志》2023年第2期188-192,共5页Medical Science Journal of Central South China

基  金:湖南省科技厅临床医疗技术创新引导项目(2020SK51703);柳州市科技计划项目(2022CAC0219)。

摘  要:目的 探讨沉默环状RNA hsa_circ_0124696(circROBO1)对鼻咽癌CNE2细胞侵袭与肺转移的影响机制。方法 qRT-PCR检测鼻咽癌及癌旁组织中circROBO1表达。采用干扰小RNA(siRNA)沉默鼻咽癌CNE2细胞中circROBO1的表达,Transwell及HE染色检测circROBO1对CNE2细胞迁移能力和体内肺转移的影响。TargetScan在线软件预测circROBO1下游miR-217与下游靶基因KRAS的靶向结合位点,双荧光素酶报告基因实验验证两者之间的靶向调控关系。蛋白免疫印迹检测siRNA沉默CNE2细胞中circROBO1表达对KRAS的影响。结果 鼻咽癌组织中circROBO1表达高于癌旁组织(P<0.05)。与转染si-circNC的对照组相比,si-circROBO1组鼻咽癌CNE2细胞侵袭与体内肺转移能力均显著降低(P<0.05)。circROBO1下游miR-217与KRAS之间存在靶向结合位点,并且circROBO1可影响KRAS的蛋白和mRNA表达量。结论 沉默circROBO1通过miR-217下调KRAS抑制鼻咽癌CNE2细胞侵袭与肺转移。Aim To explore the mechanism of silencing cyclic RNA hsa_circ_0124696(circROBO1)invasion and pulmonary metastasis of nasopharyngeal carcinoma CNE2 cells.Methods The qRT-PCR experiments detected the expression of circROBO1 in nasopharyngeal carcinoma and adjacent tissues.small interfering RNA(siRNA)was used to silence the expression of circROBO1 in CNE2 cells of nasopharyngeal carcinoma,Transwell and HE staining experiments to detect the effect of circROBO1 on the migration ability of CNE2 cells and pulmonary metastasis in vivo.The target binding site of the downstream miR-217 of circROBO1 and the downstream target gene KRAS was predicted by TargetScan online software,and the targeting regulatory relationship between the two was verified by double luciferase reporter gene experiments.Western blotting assays to detect the effect of siRNA silencing on the expression of circROBO1 on KRAS in CNE2 cells.Results The expression level of circROBO1 in nasopharyngeal carcinoma tissue was higher than that in the paracancerous tissue(P<0.05).Compared with the control group transfected with si-control,the invasion and in vivo lung metastasis abilities of CNE2 cells of nasopharyngeal carcinoma in the si-circROBO1 group were significantly reduced(P<0.05).There is a targeted binding site between miR-217 downstream of circROBO1 and KRAS,and circROBO1 can affect the protein and mRNA expression of KRAS.Conclusion Silencing circROBO1 inhibits the invasion and pulmonary metastasis of nasopharyngeal carcinoma CNE2 cells by down-regulating KRAS level through miR-217.

关 键 词:鼻咽癌 circROBO1 miR-217 KRAS 侵袭 肺转移 

分 类 号:R739.2[医药卫生—肿瘤]

 

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