摘要
目的探讨环状RNA(circRNA)hsa_circ_0102049调控结直肠癌(colorectal cancer,CRC)细胞增殖和转移能力的机制。方法从GEO数据库下载GSE126094数据集并通过R软件筛选差异表达的circRNA,构建hsa_circ_0102049的小干扰RNA(small interfering RNA,siRNA)以敲低hsa_circ_0102049的表达量并通过RT-qPCR、CCK-8、Transwell和Western blot法等检测转染后CRC细胞的增殖和转移能力变化并探寻相关机制。结果在GSE126094数据集中,hsa_circ_0102049在癌组织中表达量显著高于癌旁组织(P<0.001)。在HCT116和DLD-1细胞中干扰hsa_circ_0102049后,细胞的增殖和迁移侵袭能力减弱(P<0.05),Western blot法检测结果显示,E-cadherin的表达量升高而N-cadherin和Vimentin的表达量降低(P<0.05)。为了探寻hsa_circ_0102049的作用机制,数据库分析筛选出可以与hsa_circ_0102049结合的小RNA(microRNA,miRNA)miR-96-5p以及miR-96-5p的下游靶标AK3,并且敲低hsa_circ_0102049引起miR-96-5p的升高和AK3的降低(P<0.05)。结论在CRC细胞中,高表达的hsa_circ_0102049通过miR-96-5p/AK3轴促进上皮-间质转化(epithelial-mesenchymal transformation,EMT)从而增强细胞的增殖和转移。
Objective To investigate the function and mechanism of circRNA,hsa_circ_0102049,in regulating the proliferation and metastasis of colorectal cancer(CRC)cells.Methods GSE126094dataset was downloaded from GEO database and differently expressed circRNAs were screened by R software.The small interfering RNA(siRNA)of hsa_circ_0102049 was constructed to knockdown the expression level of hsa_circ_0102049,RT-qPCR,CCK-8,Transwell and Western blot experiments were conducted to detected the proliferation and metastasis of CRC cells after transfection and explore the related mechanism.Results In the GSE126094 dataset,hsa_circ_0102049 was significantly higher expressed in cancer tissues than in para-cancer tissues(P<0.001).After knocking down hsa_circ_0102049,the proliferation,migration and invasion ability of HCT116 and DLD-1 cells were reduced(P<0.05),meanwhile the result of Western blot indicated that the expression of E-cadherin was increased and that of N-cadherin and Vimentin decreased(P<0.05).To find the mechanism of hsa_circ_0102049 in CRC cells,related microRNA(miRNA)miR-96-5p that could bind hsa_circ_0102049 and its downstream AK3 were screened from databases,and hsa_circ_0102049 knockdown led to the increase of miR-96-5p and the decrease of AK3(P<0.05).Conclusion High expression of hsa_circ_0102049 promotes epithelial-mesenchymal transformation(EMT)through miR-96-5p/AK3 axis,thereby enhancing the proliferation and metastasis of CRC cells.
作者
刘胜楠
陈卓
LIU Shengnan;CHEN Zhuo(Department of Gastroenterology,Affiliated Hospital of Xuzhou Medical University,Jiangsu 221000,China)
出处
《医学研究杂志》
2022年第11期154-159,共6页
Journal of Medical Research