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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
- LI Guobin, ZHANG Zhancheng, WANG Xinyan
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J Otolaryngol Ophthalmol Shandong Univ. 2018, 32(4):
53-57.
doi:10.6040/j.issn.1673-3770.0.2017.301
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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.