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microRNA-124高表达对甲状腺乳头状癌细胞系K1侵袭、迁移的影响及其机制 预览

Effects of high expression of microRNA-124 on invasion and migration of thyroid papillary carcinoma cell line K1
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摘要 目的观察微小RNA 124(microRNA-124)高表达对甲状腺乳头状癌(TPC)细胞系K1侵袭、迁移的影响,并探讨其作用机制。方法取对数生长期K1细胞,分为三组,观察组用促进microRNA-124表达的microRNA-124mimics转染,对照组转染microRNA无关序列,空白组不做任何处理。培养24 h,Transwell试验观测三组侵袭数,采用细胞划痕试验观测三组迁移细胞数,采用Real-time PCR法检测细胞含IQ基元GTP酶激活蛋白1(IQGAP1)mRNA及蛋白、microRNA-124,采用荧光素酶报告基因试验检测三组细胞中含有IQGAP1基因3'非翻译区(UTR)报告质粒的荧光素酶活性。结果培养24 h时观察组、对照组、空白组细胞侵袭数分别为(104.32±11.21)、(304.27±20.83)、(299.38±18.32)个,观察组细胞侵袭数低于对照组和空白组(P均〈0.05)。培养24 h后,观察组、对照组、空白组细胞迁移数分别为(62.16±3.21)、(104.27±6.83)、(99.38±5.32),观察组划痕愈合百分比低于对照组和空白组(P均〈0.05)。观察组细胞IQGAP1 mRNA及蛋白相对表达量均低于对照组、空白组(P均〈0.05);观察组microRNA-124相对表达量均高于对照组、空白组(P均〈0.05)。培养48 h观察组、对照组、空白组基因质粒的相对荧光素酶活性分别为0.24±0.06、1.06±0.15、1.10±0.17,观察组相对荧光素酶活性低于对照组和空白组(P均〈0.05)。结论 microRNA-124高表达可抑制K1细胞的侵袭、迁移,其机制可能为microRNA-124下调IQGAP1表达。 Objective To observe the effects of high expression of microRNA-124 on the invasion and migration of thyroid papillary carcinoma(TPC) cell line K1 and to discuss its mechanism. Methods K1 cells in the logarithmic phase were divided into three groups. The observation group was transfected with microRNA-124 mimics which promoted the expression of microRNA-124,the control group was transfected with microRNA independent sequence,and the blank group was not treated. After 24-hour culture,Transwell assay was used to observe the invasion number of three groups; the migration cells were detected by Scratch test; the IQ motif-containing GTPase-activating protein 2(IQGAP1) mRNA and protein,and microRNA-124 in cells were analyzed by the real-time PCR; Luciferase reporter assay was applied to detect the luciferase activity of the reporting plasmid containing the 3-Untranslated Region(UTR) of IQGAP1 gene. Results After24-hour culture,the number of invasion cells of the observation group,the control group and the blank group was 104. 32 ±11. 21,304. 27 ± 20. 83 and 299. 38 ± 18. 32,respectively. The number of invasion cells in the observation group was lower than that in the control group and the blank group(all P〈0. 05). After 24 h culture,the migration number of cells in the observation group,the control group and the blank group was 62. 16 ± 3. 21,104. 27 ± 6. 83,and 99. 38 ± 5. 32,respectively. Healing percentage in the observation group was lower than that in the control group and the blank group(P〈0. 05). The relative expression levels of IQGAP1 RNA and protein in the observation group was lower than those in the control group and the blank group(both P〈0. 05). The relative expression level of microRNA-124 in the observation group was higher than that in the control group and the blank group(P〈0. 05). After 48 h culture,the relative luciferase activity in the observation group,the control group and the blank group was 0. 24 ± 0. 06,1. 06 ± 0. 15 and 1. 10 ± 0. 17,respectively,
作者 肖成华 阚捷 李海燕 赞梅 王新国 XIAO Chenghua, KAN Jie, LI Haiyan, ZAN Mei, WANG Xinguo (Qbtghai People's Hospital, Xining 810007, China)
机构地区 青海省人民医院
出处 《山东医药》 2018年第31期44-47,共4页 Shandong Medical Journal
基金 青海省基础研究计划资助项目(2016-K-9P12)
关键词 微小RNA microRNA-124 甲状腺肿瘤 甲状腺乳头状癌 细胞侵袭 细胞迁移 含IQ基元GTP酶激活蛋白1 microRNA microRNA-124 thyroid carcinoma thyroid papillary- carcinoma cell invasion cell migration IQ motif-containing GTPase-activating protein 2
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