| 马永红,同娟.敲低 Ⅰ型胶原 α1链基因抑制鼻咽癌细胞增殖、迁移和侵袭的研究[J].安徽医药,2021,25(4):744-747. |
| 敲低 Ⅰ型胶原 α1链基因抑制鼻咽癌细胞增殖、迁移和侵袭的研究 |
| Inhibition of proliferation, migration and invasion of nasopharyngeal carcinoma cells by knocking down COL1A1 gene |
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| DOI：10.3969/j.issn.1009-6469.2021.04.027 |
| 中文关键词: 鼻咽肿瘤 胶原 Ⅰ型 增殖 钙黏蛋白 波形蛋白 |
| 英文关键词: Nasopharyngeal neoplasms Collagen type I Proliferation Cadherin Vimentin |
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| 中文摘要: |
| 目的研究 Ⅰ型胶原 α1链(COL1A1)基因对鼻咽癌细胞增殖、侵袭、迁移的影响。方法该研究于 2017年9月至 2018年12月完成，实时定量 PCR(qPCR)和蛋白质印迹法(Western blotting)检测 COL1A1在鼻咽癌(C666-1、CNE1、CNE2)细胞系中的表达量； CNE1细胞分 m为对照组、阴性对照(si-NC组)、敲低 COL1A1(si-COL1A1)组，通过 Lipofectamine 2000将siRNA COL1A1质粒转染至鼻咽癌 CNE1细胞中，对照组细胞正常培养， si-NC组细胞转染阴性对照； qPCR和Western blotting检测转染效果，噻唑蓝(MTT)、 Transwell实验分别检测细胞增殖、侵袭、迁移； Western blotting检测钙黏蛋白 E(E-cadherin)、波形蛋白(Vimentin)钙黏蛋白 N(N-cadherin)水平。结果 COL1A1 mRNA［(3.565±0.341)(4.253±0.410)(3.968±0.378)］和蛋白表达量［(2.214±0.216)(、3.152±0.305)(2.519±0.221)］在鼻咽癌(C666-1、CNE1、CNE2)细胞系中表达量增加(P<0.05)； CNE1细胞转染 siRNA COL1A1质粒能显著下调 COL1A1 mRNA(0.352±0.045)和蛋白表达量(0.245±0.232)(P<0.05)；敲低 COL1A1的表达量抑制 CNE1细胞增殖(0.536±0.059)(0.665±0.067)(0.893±0.081)(P<0.05)降低其侵袭(52.895±5.635)迁移(113.254±13.279)能力(P<0.05)，上调 E-cadherin蛋白水平(2.152±0.223)(P<0.05)，下调 Vimentin(，0.362±0.045)、N-cadherin(、0.423±0.043)蛋白水平(P<0.05)。结论 COL1A1可能通过抑制上皮细胞间充质化(EMT)促进鼻咽癌细胞增殖、侵袭、迁移。 |
| 英文摘要: |
| Objective To investigate the effect of collagen type l alphα 1 (COL1A1) gene on proliferation, invasion and migration of nasopharyngeal carcinoma cells.Methods The study was completed from September 2017 to December 2018. Quantitative polymerasechain reaction (qPCR) and Western blotting were used to detect the expression of COL1A1 in nasopharyngeal cancer cell lines (C666-1,CNE1, CNE2). CNE1 cells were randomly assigned into control group, negative control (si-NC) group and knockdown COL1A1 (siCOL1A1) group. The siRNA COL1A1 plasmid was transfected into CNE1 cells by Lipofectamine 2000, and the control group were cultured normally, while those in si-NC group were transfected into negative control group. The transfection effect was detected by qRT-PCR and Western blotting. The cells proliferation, invasion and migration were detected by MTT and Transwell assay. Western blottingwas used to detect the expression of E-cadherin, Vimentin, N-cadherin.Results The expression of COL1A1 mRNA［(3.565±0.341),(4.253±0.410),(3.968±0.378)］and protein expressions［(2.214±0.216),(3.152±0.305),(2.519±0.221)］was significantly increased in nasopharyngeal cancer cell lines (C666-1, CNE1, CNE2) (P<0.05). The siRNA COL1A1 plasmid could down-regulate the expression of COL1A1 (0.352±0.045) gene and (0.245±0.232) protein (P<0.05). Knocking down the expression of COL1A1 inhibited the proliferation［(0.536±0.059),(0.665±0.067),(0.893±0.081)］of CNE1 cells (P<0.05), decreased the invasion (52.895±5.635) and migration (113.254± 13.279) ability (P<0.05), increased the level of E-cadherin (2.152±0.223) protein (P<0.05), and decreased the levels of Vimentin (0.362± 0.045) and N-cadherin (0.423±0.043) protein (P<0.05).Conclusion COL1A1 may promote the proliferation, invasion and migration of nasopharyngeal carcinoma cells by inhibiting epithelial-mesenchymal transition (EMT). |
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