| 张志勇,潘 妍,赵 艳,任牡丹,李雅睿,卢桂芳,和水祥.下调UCA1通过靶向miR-23b及其下游靶基因抑制胃癌细胞的增殖和转移[J].,2021,(22):4238-4243 |
| 下调UCA1通过靶向miR-23b及其下游靶基因抑制胃癌细胞的增殖和转移 |
| Down-regulation of UCA1 Inhibits the Proliferation and Metastasis of Gastric Cancer Cells by Targeting miR-23b and Its Downstream Target Genes |
| 投稿时间：2021-03-21 修订日期：2021-04-17 |
| DOI：10.13241/j.cnki.pmb.2021.22.008 |
| 中文关键词: 胃癌 尿路上皮癌相关基因1 miR-23b-3p 增殖 转移 |
| 英文关键词: Gastric cancer Urothelial carcinoma-associated 1 MiR-23b-3p Proliferation Metastasis |
| 基金项目:国家自然科学基金项目（81900489）；陕西省自然科学基础研究项目（2020JM-381）；陕西省重点研发计划：重点产业创新链（群）项目（2021ZDLSF02-06） |
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| 中文摘要: |
| 摘要 目的：探讨长链非编码RNA尿路上皮癌相关基因1（UCA1）调控胃癌细胞增殖和转移的分子机制。方法：将人胃癌细胞株SGC7901分为：对照组、siRNA-NC组、siRNA-UCA1组、inhibitor-NC组和miR-inhibitor组、si-UCA1+inhibitor-NC组和si-UCA1+miR-inhibitor组。对SGC7901细胞分别转染siRNA-UCA1及阴性对照（siRNA-NC）、miR-inhibitor及阴性对照（inhibitor-NC），未转染的细胞作为对照组。通过RT-qPCR检测细胞中UCA1和miR-23b-3p的水平。通过CCK-8法、伤口愈合实验和Transwell实验评价细胞的增殖、迁移和侵袭能力。通过Western blot分析细胞中IL6R、BCL2和HSP90B1蛋白的表达。使用pcDNA-UCA1/pcDNA-NC与pGL3-miR-23b-3p-WT/pGL3-miR-23b-3p-Mut共转染细胞，通过双荧光素酶报告实验验证UCA1与miR-23b-3p的靶向关系。结果：细胞培养48 h和72 h后，与对照组比较，siRNA-UCA1组的细胞活力分别降低了31.58%和31.40%（P<0.05）。与对照组比较，siRNA-UCA1组的细胞迁移率[(61.46±5.43)% vs (23.16±3.17)%]、侵袭细胞数量（109.17±9.66 vs 50.83±6.96）、IL6R、BCL2和HSP90B1的蛋白相对表达量均显著降低，而miR-23b-3p相对表达量升高（P<0.05）。与pGL3-miR-23b-3p-WT共转染后，与pcDNA-NC组比较，pcDNA-UCA1组的相对荧光酶活性降低了66.12%（P<0.05）。与si-UCA1+inhibitor-NC组比较，si-UCA1+miR-inhibitor组的细胞活力、细胞迁移率、侵袭细胞数量、IL6R、BCL2和HSP90B1的蛋白相对表达量均显著升高（P<0.05）。结论：下调UCA1通过靶向miR-23b-3p及其下游基因IL6R、BCL2和HSP90B1来抑制胃癌细胞的增殖和转移。 |
| 英文摘要: |
| ABSTRACT Objective: To explore the molecular mechanism of long non-coding RNA urothelial carcinoma-associated 1 (UCA1) regulating the proliferation and metastasis of gastric cancer cells. Methods: The human gastric cancer cell line SGC7901 was divided into the following groups: control group, siRNA-NC group, siRNA-UCA1 group, inhibitor-NC group, miR-inhibitor group, si-UCA1+inhibitor-NC group, si-UCA1+miR-inhibitor group. SGC7901 cell was transfected with siRNA-UCA1 or negative control (siRNA-NC), miR-inhibitor or negative control (inhibitor-NC) respectively, and the untransfected cells were used as control group. The levels of UCA1 and miR-23b-3p in the cells were detected by RT-qPCR. The CCK-8 method, wound healing experiment and Transwell experiment were used to evaluate cell proliferation, migration and invasion ability. The expression of IL6R, BCL2 and HSP90B1 protein in the cells was analyzed by Western blot. Cells were co-transfected with pcDNA-UCA1/pcDNA-NC and pGL3-miR-23b-3p-WT/pGL3-miR- 23b-3p-Mut, and the target relationship between UCA1 and miR-23b-3p was verified by dual luciferase report experiment. Results: After culturing for 48 h and 72 h, compared with Control group, the cell viability of siRNA-UCA1 group was reduced by 31.58% and 31.40%, respectively (P<0.05). Compared with Control group, the cell migration rate [(61.46±5.43)% vs (23.16±3.17)%], the number of invasive cells(109.17±9.66 vs 50.83±6.96), and the relative expression levels of IL6R, BCL2 and HSP90B1 proteins in siRNA-UCA1 group were all significant reduced, but the relative expression of miR-23b-3p significant increased (P<0.05). After co-transfection with pGL3-miR-23b-3p-WT, compared with pcDNA-NC group, the relative luciferase activity of pcDNA-UCA1 group was reduced by 66.12% (P<0.05). Compared with si-UCA1+inhibitor-NC group, cell viability, cell migration rate, number of invaded cells, relative expression of IL6R, BCL2 and HSP90B1 protein in si-UCA1+miR-inhibitor group were significantly increased (P<0.05). Conclusion: Down-regulation of UCA1 inhibits the proliferation and metastasis of gastric cancer cells by targeting miR-23b-3p and its downstream genes IL6R, BCL2 and HSP90B1. |
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