上调miR-200b对人喉癌Hep-2细胞增殖、迁移和侵袭能力的影响

doi:10.6040/j.issn.1673-3770.0.2017.301

摘要/Abstract

摘要： 目的探讨上调miR-200b对人喉癌Hep-2细胞增殖、迁移和侵袭能力的影响。方法培养人喉癌Hep-2细胞,随机分为miR-200b模拟物组、miR-对照序列组和空白对照组,利用实时荧光定量PCR技术检测各组细胞中miR-200b表达,MTT法检测细胞增殖能力,Transwell法检测细胞迁移和侵袭能力,Western blotting法检测各组细胞中E-cadherin、N-cadherin、β-catenin蛋白表达。结果与空白对照组和miR-对照序列组相比,miR-200b模拟物组细胞中miR-200b相对表达量显著升高,差异有统计学意义(F=70.766, P<0.001);与空白对照组［(0.22±0.05)、(0.45±0.07)、(0.59±0.06)、(0.72±0.08)和(0.85±0.07)］和miR-对照序列组［(0.24±0.06)、(0.46±0.08)、(0.61±0.07)、(0.70±0.05)和(0.84±0.06)］相比,miR-200b模拟物组细胞24、48、72、96 h时吸光度A值［(0.21±0.04)、(0.29±0.06)、(0.40±0.04)、(0.53±0.07)和(0.58±0.05)］均降低,差异均有统计学意义(F=0.134, P=0.876; F=6.449, P=0.010; F=37.299, P<0.001; F=5.352, P=0.018; F=29.921, P<0.001);与空白对照组［(140.2±2.3)、(127.9±6.0)］和miR-对照序列组［(141.0±1.2)、(130.4±7.4)］相比,miR-200b模拟物组迁移细胞数和侵袭细胞数［(98.5±2.4)、(90.9±2.8)］均减少,差异均有统计学意义(F=845.523, P<0.001; F=88.859, P<0.001);与空白对照组［(0.35±0.07)、(0.54±0.10)、(0.76±0.12)］和miR-对照序列组［(0.24±0.03)、(0.60±0.14)、(0.65±0.24)］相比,miR-200b模拟物组细胞中E-cadherin蛋白相对表达量(0.54±0.14)升高,而N-cadherin、β-catenin蛋白相对表达量［(0.31±0.10)、(0.35±0.06)］降低,差异均有统计学意义(F=17.287, P<0.001; F=10.083, P=0.002; F=10.434, P=0.001)。结论上调喉癌细胞中miR-200b基因表达可减少细胞增殖,抑制细胞迁移和侵袭能力,其机制可能与抑制上皮-间质转化过程有关。

关键词: 上皮-间质转化, 细胞增殖, 喉癌, miR-200b, 细胞侵袭

Abstract: Objective We investigated the effects of up-regulation of miR-200b on the proliferation, migration, and invasion of human laryngeal cancer Hep-2 cells. Methods Human laryngeal carcinoma Hep-2 cells were cultured and randomly divided into the miR-200b mimic group, miR-control sequence group, and blank control group. The expression of miR-200b was detected by real-time fluorescence quantitative PCR. The MTT assay was used to detect cell proliferation. The Transwell method was used to detect cell migration and invasion. The expression of E-cadherin, N-cadherin, and β-catenin proteins was detected by western blotting. Results Compared to the blank control group and miR-control sequence group, the relative expression level of miR-200b in the miR-200b mimic group was significantly increased(F=70.766, P<0.001). Compared to the blank control group ［(0.22±0.05),(0.45±0.07),(0.59±0.06),(0.72±0.08), and(0.85±0.07)］ and miR-control sequence group ［(0.24±0.06),(0.46±0.08),(0.61±0.07),(0.70±0.05), and(0.84±0.06)］, the absorbance A values at 24, 48, 72, and 96 h in the miR-200b mimic group ［(0.21±0.04),(0.29±0.06),(0.40±0.04),(0.53±0.07), and(0.58±0.05)］ were decreased and the differences were statistically significant(F=0.134, 6.449, 37.299, 5.352, and 29.921, P=0.876, 0.010, 0.000, 0.018, and 0.000, P<0.05). Compared to the blank control group ［(140.2±2.3),(127.9±6.0)］ and miR-control sequence group ［(141.0±1.2),(130.4±7.4)］, the number of migrating cells and number of invasive cells in the miR-200b mimic group ［(98.5±2.4),(90.9±2.8)］ were decreased and the differences were statistically significant(F=845.523, 88.859, P both <0.001). Compared to the blank control group ［(0.35±0.07),(0.54±0.10),(0.76±0.12)］ and miR-control sequence group ［(0.24±0.03),(0.60±0.14),(0.65±0.24)］, the relative expression level of E-cadherin protein in miR-200b mimic group cells(0.54±0.14)was increased, whereas the relative expression levels of N-cadherin and β-catenin proteins ［(0.31±0.10),(0.35±0.06)］ were decreased; these differences were statistically significant(F=17.287, 10.083, 10.434, P<0.001, 0.002, 0.001). Conclusion Up-regulation of miR-200b gene expression in laryngeal squamous cell carcinoma may reduce cell proliferation and inhibit cell migration and invasion. The mechanism may be related to inhibition of the epithelial mesenchymal transition process.

Key words: miR-200b, Epithelial-mesenchymal transition, Cell proliferation, Cell invasion, Laryngeal carcinoma

中图分类号:

R739.6

李国彬,张占成,王新颜. 上调miR-200b对人喉癌Hep-2细胞增殖、迁移和侵袭能力的影响[J]. 山东大学耳鼻喉眼学报, 2018, 32(4): 53-57.

LI Guobin, ZHANG Zhancheng, WANG Xinyan. Up-regulation of miR-200b may inhibit epithelial mesenchymal transition process to prevent the proliferation, migration, and invasion of human laryngeal cancer Hep-2 cells[J]. J Otolaryngol Ophthalmol Shandong Univ, 2018, 32(4): 53-57.

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