MiR－181a和miR－181b靶向调控FUT1对结直肠癌进展的影响  

MiR-181a and miR-181b influence proliferation, migration, invasion and angiogenesis via targeting FUT1 in colorectal cancer

作　　者：周杰[1] 郑远明[1] 沈宗坤 马晓露[3] 李艳菊 Zhou Jie;Zheng Yuanming;Shen Zongkun;Ma Xiaolu;Li Yanju(Department of Clinical Laboratory Medicine,Dalian Friendship Hospital,Dalian 116001,China)

机构地区：[1]大连市友谊医院检验科,116001 [2]阜新中心医院检验科 [3]大连医科大学附属第一医院检验科 [4]中国医科大学附属盛京医院大连医院检验科

出　　处：《中华检验医学杂志》2018年第11期841-846,共6页Chinese Journal of Laboratory Medicine

摘　　要：目的通过研究miR－181a及miR－181b与岩藻糖基转移酶FUT1的相关性，阐明miR－181a和miR－181b靶向调控FUT1在结直肠癌转移中的功能机制。方法收集2014年3月至2016年1月大连医科大学附属第一医院经手术切除的结直肠癌及其癌旁组织共32对组织标本，男性18例，女性14例；采用RT－PCR方法检测了32对结直肠癌患者癌组织、癌旁组织、结直肠癌患者和健康人血清及结直肠癌高转移细胞株SW620、低转移细胞株SW480中miR－181a和miR－181b的表达。通过皮尔森＂Pearson＂相关曲线得出FUT1与miR－181a、miR－181b表达相关趋势。通过生物信息学网站TargetScan、microRNA.org和Starbase v2.0预测及双荧光素酶实验报告验证miR－181a、miR－181b与FUT1的靶向关系；通过CCK8，划痕实验，transwell小室及血管形成进一步检测SW480、SW620细胞中miR－181a和miR－181b表达调控对肿瘤细胞增殖、迁移、侵袭及血管形成的影响。两独立样本间的比较采用t检验，多个样本间的比较采用单因素方差分析，相关分析采用Pearson相关系数分析。结果miR－181a、miR－181b在结直肠癌组织中的表达明显低于癌旁组织（3.12±1.88 vs 6.44±2.32，t＝11.74; 3.16±1.77 vs 5.52±2.45, t＝3.24，P均〈0.05）；miR－181a、miR－181b在结直肠癌患者血清的表达量明显低于健康人血清中的表达量（1.32±0.25，2.57±0.48，t＝10.26；0.91±0.14，1.63±0.29，t＝5.19；P均〈0.05）；在结直肠癌细胞株SW480和SW620中的表达明显低于正常结直肠上皮细胞株FHC[（0.65±0.10、0.50±0.09）vs 1.0, （0.60±0.12、0.42±0.03）vs 1.0; t＝3.08，P均〈0.05]; FUT1在结直肠癌组织及SW620中高表达（t＝5.23，P〈0.05）。TargetScan、microRNA.org和Starbase v2.0软件预测FUT1与miR－181a、miR－181b具有靶向结合位点，双荧光素酶实验验证FUT1为miR－181a、miR－181b的共同靶基因。特异性上调SW620细胞中miR－181a、miR－181b，细�ObjectiveTo investigate the correlation of miR-181a and miR-181b with fucosyltransferase FUT1, the functional mechanism was elucidated in a colorectal cancer （CRC）. MethodsIt collected 32 pairs of tissue samples, 18 males and 14 females in the first affiliated hospital of Dalian Medicinal University, from March 2014 to January 2016. The expression of miR-181a and miR-181b was detected by RT-PCR in CRC tissues, adjacent tissues, serum of colorectal cancer patients and healthy people, and CRC cell lines SW620 and SW480 with differently metastatic ability. The relationship of FUT1 and miR-181a, miR-181b expression were verificated by Pearson′s correlation curve. FUT1 was identified the target of miR-181a and miR-181b by Network prediction softwares （TargetScan Human 7.1, microRNA.org and Starbase v2.0） and luciferase assay. The effects of miR-181a and miR-181b expression on the proliferation, migration, invasion and angiogenesis of SW480 and SW620 cells were further detected by CCK8, wound healing, transwell and tube foramtion assays. T-test was used for comparison between two independent samples, and one-way anova was used for comparison between multiple samples. Pearson correlation coefficient was used for correlation analysis. ResultsThe levels of miR-181a and 181b in CRC tissues were much lower than in tumor-adjacent tissues （3.12±1.88 vs 6.44±2.32, t=11.74; 3.16±1.77 vs 5.52±2.45, t=3.24 ; P〈0.05）. The levels of miR-181a and 181b in serum of colorectal cancer patients were much lower than in healthy people（1.32±0.25, 2.57±0.48, t=10.26; 0.91±0.14, 1.63±0.29, t=5.19; P〈0.05）. The levels of miR-181a and miR-181b in SW620, SW480 CRC cells were detected to be much lower than in normal colorectal epithelial cells[（0.65±0.10, 0.50±0.09）vs 1.0; （0.60±0.12, 0.42±0.03）vs 1.0; t=3.08, P〈0.05]. FUT1 was highly expressed in CRC tissues and SW620 （t=5.23, P〈0.05）. Based on the network prediction softwares and luciferase assays, FUT1 was the common target of miR-181a and mi

关 键 词：结直肠肿瘤 岩藻糖基转移酶类 微RNAS 细胞增殖 肿瘤侵润 新生血 管化 病理性 

分 类 号：R735.34[医药卫生—肿瘤]