| 尚念红.微小 RNA-7-5p靶向调节锌指蛋白核转录因子 4基因抑制口腔鳞状细胞癌细胞增殖和诱导凋亡的体外研究[J].安徽医药,2021,25(12):2470-2474. |
| 微小 RNA-7-5p靶向调节锌指蛋白核转录因子 4基因抑制口腔鳞状细胞癌细胞增殖和诱导凋亡的体外研究 |
| Inhibition of proliferation and induction of apoptosis in oral squamous cell carcinoma cells by miR-7-5p targeting KLF4 gene |
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| DOI：10.3969/j.issn.1009-6469.2021.12.032 |
| 中文关键词: 口腔肿瘤 癌，鳞状细胞 微小 RNA-7-5p 锌指蛋白核转录因子 4基因 增殖 凋亡 |
| 英文关键词: Mouth neoplasms Carcinoma, squamous cell MiR-7-5p KLF4 gene Proliferation Apoptosis |
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| 中文摘要: |
| 目的探讨微小 RNA(miR)-7-5p对口腔鳞状细胞癌细胞增殖、凋亡及锌指蛋白核转录因子 4(KLF4)基因表达调控的影响。方法研究起止时间为 2018年 3—10月。实时荧光定量逆转录聚合酶链反应( qRT-PCR)检测 miR-7-5p在人正常口腔黏膜成纤维细胞 hOMF及口腔鳞状细胞癌细胞 SCC25、Cal127和 Tca8113中的表达量。将 miR-7-5p模拟物( miR-7-5p mimics)及阴性对照序列( mimics Control)分别转染至 SCC25细胞，分为三组: Control组(未转染的 SCC25细胞)、 NC组(转染 mimics Con. trol的 SCC25细胞)、 miR-7-5p组(转染 miR-7-5p mimics的 SCC25细胞)。以 qRT-PCR检测 miR-7-5p在 SCC25细胞中的转染效果； MTT法检测 miR-7-5p对 SCC25细胞增殖的影响；流式细胞术检测 miR-7-5p对 SCC25细胞凋亡的影响；通过双荧光素酶报告基因实验、 qRT-PCR和蛋白质印迹法( Western blotting)检测 miR-7-5p对 KLF4的靶向关系。结果 miR-7-5p在 3种口腔鳞状细胞癌细胞 SCC25、Cal127和 Tca8113中的表达量［(0.21±0.02)、(0.43±0.04)、(0.48±0.05)］明显低于人正常口腔黏膜成纤维细胞 hOMF(1.00±0.09)(P<0.05)；转染 miR-7-5p mimics能够上调 SCC25细胞中 miR-7-5p的表达( P<0.05)； miR-7-5p过表达能够明显抑制 SCC25细胞增殖能力［Control组、 NC组 72 h吸光度( 1.43±0.13)、(1.46±0.15)比 miR-7-5p组( 0.81±0.09)］(P<0.05)，并诱导细胞凋亡［Control组、 NC组凋亡率( 9.48±2.65)%、(10.21±3.02)%比 miR-7-5p组( 24.41±8.15)%］(P<0.05)；双荧光素酶报告基因实验结果表明 miR-7-5p过表达可抑制 KLF4-Wt报告基因载体细胞相对荧光活性； qRT-PCR和 Western blotting进一步证 |
| 英文摘要: |
| Objective To investigate the effect of microRNA(miR)-7-5p on proliferation, apoptosis and regulation of zinc finger pro.tein nuclear factor 4 (KLF4) gene expression in oral squamous cell carcinoma cells.Methods The study started from March 2018 and ended in October 2018. Real-time fluorescence quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to de. tect the expressions of miR-7-5p in human normal oral mucosal fibroblast hOMF and oral squamous cell carcinoma cells SCC25, Cal127 and Tca8113; miR-7-5p mimics and mimics Control were transfected into SCC25 cells, respectively. The cells were assigned in.to the following three groups: Control group (untransfected SCC25 cells), NC group (SCC25 cells transfected with mimics Control) andmiR-7-5p group (SCC25 cells transfected with miR-7-5p mimics). The transcriptional effect of miR-7-5p in SCC25 cells was detected by qRT-PCR. The proliferation of SCC25 cells by miR-7-5p was detected by MTT assay. The effect of miR-7-5p on apoptosis of SCC25 cells was detected by flow cytometry. The targeting relationship of miR-7-5p to KLF4 was detected by double luciferase reporter gene assay, qRT-PCR and Western blotting.Results The expressions of miR-7-5p in three oral squamous cell carcinoma cells SCC25,Cal127 and Tca8113 were significantly lower [(0.21±0.02), (0.43±0.04), (0.48±0.05)] than that in normal oral mucosa fibroblast hOMF(1.00±0.09) (P<0.05). Transfection of miR-7-5p mimics significantly up-regulated the expression of miR-7-5p in SCC25 cells (P<0.05). Overexpression of miR-7-5p significantly inhibited the proliferation [(72 h absorbance: Control group (1.43±0.13), NC group (1.46± 0.15) vs. miR-7-5p group (0.81±0.09)] of SCC25 cells and induced apoptosis [apoptosis rate: Control group (9.48±2.65)%, NC group (10.21±3.02)% vs. miR-7-5p group (24.41±8.15)%] (P<0.05). Dual luciferase reporter gene assay results showed that miR-7 -5p overex. pression significantly inhibited the relative fluorescence activity of KLF4-Wt reporter vector cells. Results of qRT-PCR and Western blotting further confirmed that miR-7-5p can target KLF4 mRNA and protein expression.Conclusion MiR-7-5p directly targets theexpression of KLF4 gene to inhibit the proliferation of oral squamous cell carcinoma SCC25 cells and induce apoptosis. |
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