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miR-451靶向调控c-Myc抑制多发性骨髓瘤RPMI-8226细胞的增殖侵袭和迁移

miR-451 inhibits malignant progression of multiple myeloma RPMI-8226 cells by targeting c-Myc

摘要:

目的:探讨miR-451对多发性骨髓瘤RPMI-8226细胞增殖、侵袭和迁移的影响及其作用机制。方法:将体外培养的RPMI-8226细胞分为空白对照组(未转染)、阴性对照组(转染miR-451模拟物阴性对照)和miR-451组(转染miR-451模拟物),采用实时定量PCR检测各组细胞中miR-451的表达,采用四甲基偶氮唑蓝法和克隆形成实验检测细胞的增殖情况,Transwell小室实验检测细胞的侵袭和迁移情况,Western blot检测c-Myc、基质金属蛋白酶2(MMP-2)和MMP-9蛋白的表达,双荧光素酶报告基因实验检测miR-451和c-Myc的靶向关系。结果:miR-451组和空白对照组细胞中miR-451的表达水平分别为2.85±0.27和1.02±0.06,细胞存活率分别为(47.28±3.15)%和(93.65±6.52)%,克隆形成率分别为(15.03±1.34)%和(28.48±2.12)%,侵袭细胞数分别为(86.65±5.58)个和(135.47±9.85)个,迁移细胞数分别为(106.36±6.48 )个和(165.28±11.05)个,c-Myc蛋白的表达水平分别为0.35±0.03和0.66±0.05,MMP-2蛋白的表达水平分别为0.20±0.02和0.48±0.03,MMP-9蛋白的表达水平分别为0.28±0.03和0.59±0.06,差异均有统计学意义(均 P<0.05)。阴性对照组细胞的miR-451表达水平、细胞存活率、克隆形成率、侵袭细胞数、迁移细胞数、c-Myc蛋白的表达水平、MMP-2蛋白的表达水平和MMP-9蛋白的表达水平分别为0.94±0.05、(95.16±5.04)%、(27.55±2.26)% 、(128.96±8.32)个、(158.65±8.76)个、0.68±0.06、0.51±0.03和0.54±0.03,与空白对照组比较,差异均无统计学意义(均 P>0.05)。双荧光素酶报告基因实验显示,c-Myc为miR-451的潜在靶基因。 结论:miR-451可通过靶向c-Myc抑制RPMI-8226细胞的增殖、侵袭和迁移。

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abstracts:

Objective:To investigate the effects of microRNA-451 on proliferation, invasion and migration of multiple myeloma RPMI-8226 cells and its mechanism.Methods:RPMI-8226 cells cultured in vitro were divided into blank control group (untransfected), negative control (NC) group and miR-451 mimic transfected (miR-451) group. The expression of miR-451 was detected by real-time quantitative PCR (qRT-PCR), cell proliferation was detected by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) array and clone formation experiment, cell invasion and migration were detected by Transwell, and the expressions of c-Myc, MMP-2 and MMP-9 proteins were detected by western blot. The targeting relationship between miR-451 and c-Myc was detected by double luciferase reporter gene assay. Results:Compared to the blank control group, the expression level of miR-451 was increased (2.85±0.27 vs 1.02±0.06), while the cell survival rate [(47.28±3.15)% vs (93.65±6.52)%], cloning formation rate [(15.03±1.34)% vs (28.48±2.12)%], invasive cell number (86.65±5.58 vs 135.47±9.85), migrating cell number (106.36±6.48 vs 165.28±11.05) and the expression level of c-Myc(0.35±0.03 vs 0.66±0.05), MMP-2 (0.20±0.02 vs 0.48±0.03) and MMP-9 (0.28±0.03 vs 0.59±0.06) protein were significantly decreased in the miR-451 group ( P<0.05). In the negative control group, the expression level of miR-451, cell viability, clone formation rate, invasive cell number, migrating cell number, c-Myc protein, MMP-2 protein and MMP-9 protein were 0.94±0.05, (95.16±5.04)%, (27.55±2.26)%, (128.96±8.32) and (158.65±8.76), 0.68±0.06, 0.51±0.03, 0.54±0.03, respectively. There were no significant differences between the blank control group and the NC group ( P>0.05). Double luciferase reporter gene experiment confirmed that c-Myc was a potential target gene of miR-451. Conclusion:miR-451 can inhibit the proliferation, invasion and migration of RPMI-8226 cells by targeting c-Myc.

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作者: 鹿军 [1] 门丽杰 [1] 李弹弹 [1] 孙玲 [1] 吴希锋 [1]
作者单位: 山东第一医科大学附属济南人民医院 济南市人民医院血液科 271100 [1]
期刊: 《中华肿瘤杂志》2020年42卷7期 560-564页 MEDLINEISTICPKUCSCDCABP
栏目名称: 基础研究
DOI: 10.3760/cma.j.cn112152-20190401-00206
发布时间: 2020-09-15
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