高低淋巴结转移潜能鼻咽癌细胞与LEC共培养后目标细胞miR-92a-3p表达及其靶基因功能分析  

Expression,target genes,and function of miR-92a-3p in target cells after co-culture of high and low lymph node metastasis potential of nasopharyngeal carcinoma cells with lymphatic endothelial cells

作　　者：蒋旭 刘琦 张莹 姚茜 韦正波 谢莹 JIANG Xu;LIU Qi;ZHANG Ying;YAO Xi;WEI Zhengbo;XIE Ying(Guangxi Medical University,Nanning 530000,China;不详)

机构地区：[1]广西医科大学生命科学研究院,广西南宁530000 [2]广西医科大学附属肿瘤医院 [3]广西区域性高发肿瘤早期防治研究教育部重点实验室

出　　处：《山东医药》2020年第9期22-26,共5页Shandong Medical Journal

基　　金：国家自然科学基金地区科学基金项目(81760361);广西自然科学基金面上项目(2017GXNSFAA198064)。

摘　　要：目的观察高低淋巴结转移潜能的鼻咽癌(NPC)细胞系5-8F(高淋巴结转移潜能)及6-10B(低淋巴结转移潜能)与淋巴管内皮细胞(LEC)共培养后各目标细胞中miR-92a-3p的表达情况,预测其靶基因并分析其功能,探讨其对NPC淋巴结转移的影响及作用机制。方法依据Transwell上下室接种细胞的不同进行分组,LEC+5-8F组Transwell上室接种LEC、下室接种5-8F细胞,LEC+6-10B组上室接种LEC、下室接种6-10B细胞,5-8F+LEC组上室接种5-8F细胞、下室接种LEC,6-10B+LEC组上室接种6-10B细胞、下室接种LEC,作为对照的LEC组、5-8F组、6-10B组分别接种LEC、5-8F细胞、6-10B细胞于6孔板中,以共培养后的Transwell上室细胞和6孔板培养的单细胞为目标细胞。各组细胞培养48 h时,采用qRT-PCR法检测各组目标细胞中miR-92a-3p。通过4个miRNA靶基因预测网站miRDB、miRTarbase、miRWalk、TargetScan预测miR-92a-3p的靶基因并取交集,使用KOBAS3.0在线网站对共同预测的靶基因进行KEGG通路分析。结果LEC+5-8F组、LEC+6-10B组、LEC组细胞中miR-92a-3p的相对表达量分别为605.00±119.00、46.30±11.40、1.01±0.16,组间相比,P均<0.05。5-8F+LEC组、5-8F组细胞中miR-92a-3p的相对表达量分别为212.00±39.40、1.00±0.08,组间相比,P<0.05。6-10B+LEC组、6-10B组细胞中miR-92a-3p的相对表达量分别为4.14±0.87、1.02±0.24,组间相比,P<0.05。有74个基因被4个miRNA靶基因预测网站共同预测为miR-92a-3p的靶基因,富集在TGFβ、PI3k/Akt、AMPK、mToR以及钙信号通路等与肿瘤转移相关的通路上。结论高低淋巴结转移潜能的NPC细胞系与LEC共培养后目标细胞中miR-92a-3p表达均升高,NPC细胞系与LEC共培养后细胞中miR-92a-3p的表达水平与淋巴结转移潜能有关,miR-92a-3p可能是NPC淋巴结转移的潜在标志物。Objective To observe the expression of miR-92a-3p in the target cells after co-culture of high(5-8F)and low(6-10B)lymph node metastasis potential of nasopharyngeal carcinoma cells(NPC)with lymphatic endothelial cells(LEC),to predict the target genes and analyze their function,and to explore its effect on lymph node metastasis of NPC and its mechanism.Methods According to the different cells in Transwell's upper and lower chambers,we divided the cells into different groups;in the LEC+5-8F group,the upper chamber was inoculated with LEC and the lower chamber was inoculated with 5-8F cells;in the LEC+6-10B group,the cells were inoculated with LEC in the upper chamber and 6-10B cells in the lower chamber;in the 5-8F+LEC group,the cells were inoculated with 5-8F cells in the upper chamber and LEC in the lower chamber;in the 6-10B+LEC group,the cells were inoculated with 6-10B cells in the upper chamber and LEC in the lower chamber.As the controls,the cells in the LEC group,5-8F group,and 6-10B group were inoculated with LEC,5-8F cells,and 6-10B cells in 6-well plates,respectively.When cells of each group were cultured for 48 h,qRT-PCR was used to detect miR-92a-3p in target cells of each group.Four miRNA target gene prediction websites miRDB,miRTarbase,miRWalk,and TargetScan were used to predict the target genes of miR-92a-3p and we took intersections.KEGG pathway analysis of commonly predicted target genes was conducted by using KOBAS 3.0 online website.Results The relative expression levels of miR-92a-3p in cells of the LEC+5-8F group,LEC+6-10B group,and LEC group were 605.00±119.00,46.30±11.40,and 1.01±0.16,respectively,with statistically significant differences(all P<0.05).The relative expression levels of miR-92a-3p in cells of the 5-8F+LEC group and 5-8F group were 212.00±39.40 and 1.00±0.08,respectively,with statistically significant difference(P<0.05).The relative expression of miR-92a-3p in cells of the 6-10B+LEC group and 6-10B group were 4.14±0.87 and 1.02±0.24,respectively,with statistically signifi

关 键 词：微小RNA miR-92a-3p 鼻咽癌 鼻咽癌细胞系 淋巴管内皮细胞 

分 类 号：R739.62[医药卫生—肿瘤]