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投稿时间：2023-03-14 网络发布日期：2023-09-19
投稿时间：2023-03-14 网络发布日期：2023-09-19
中文摘要: 目的 探讨微小RNA(miR)-21对胰腺癌细胞增殖、凋亡及有氧糖酵解的影响及其可能机制。方法 选取人胰腺癌细胞SW1990、正常胰腺导管上皮细胞HPDE为研究对象。对SW1990细胞进行转染，分为miR-NC组（转染miR-NC）、miR-21-mimic组（转染miR-21-mimic）。MTT、克隆形成实验检测细胞增殖能力；流式细胞术检测细胞凋亡；RT-qPCR检测细胞中miR-21、M2型丙酮酸激酶（PKM2）基因mRNA表达水平；western blot检测PKM2蛋白水平；荧光素酶报告实验检测miR-21与PKM2的靶向关系。结果 SW1990细胞miR-21表达水平高于HPDE细胞（0.79±0.02 vs 0.23±0.02, t =34.290，P＜0.01）；SW1990细胞PKM2蛋白(0.41±0.03)、mRNA的表达水平（0.62±0.02）低于HPDE细胞（0.95±0.02、 1.13±0.03， t =25.940、24.500，P＜0.01）。miR-21-mimic组72 h、96 h时细胞增殖能力、细胞克隆数明显高于miR-NC组，凋亡细胞数低于miR-NC组（P＜0.05）。miR-21-mimic组细胞葡萄糖消耗量和乳酸、己糖激酶（HK）及乳酸脱氢酶（LDH）含量均明显高于miR-NC组（ t =5.992、21.340、5.643、4.008，P＜0.05）。miR-21-mimic组细胞PKM2蛋白表达水平低于miR-NC组（ t =10.070，P＜0.01）。miR-21与野生型-3′非翻译区（WT-3′UTR）-PKM2存在一定数量的互补碱基对，miR-21-mimic+WT-PKM2组的细胞荧光素酶活性低于miR-NC+WT-PKM2组（ t =15.680，P＜0.01）。结论 胰腺癌细胞miR-21高表达，miR-21过表达可能促进胰腺癌细胞增殖、有氧糖酵解，抑制细胞凋亡，其机制可能与靶向PKM2有关。
Abstract:Objective To investigate the effects of microRNA-21 (miR-21) on proliferation, apoptosis and aerobic clycolysis of pancreatic cancer cells and its possible mechanism. Methods Human pancreatic cancer cell SW1990 and normal pancreatic duct epithelial cell HPDE were taken as research objects. SW1990 cells were transfected and divided into miR-NC group (transfected with miR-NC) and miR-21-minic group (transfected with miR-21-minic). MTT and clone formation assay were used to detect cell proliferation ability. Flow cytometry was used to detect cell apoptosis. RT-qPCR was used to detect the mRNA expression levels of miR-21 and pyruvate kinase M2 (PKM2) in cells. Western blot was used to detect PKM2 protein level. The luciferase reporter assay was used to detect the targeting relationship between miR-21 and PKM2. Results The expression level of miR-21 in SW1990 cells was higher than that in HPDE cells (0.79±0.02 vs 0.23±0.02, t =34.290, P＜0.01), while the expression levels of PKM2 protein (0.41±0.03) and mRNA (0.62±0.02) in SW1990 cells were lower than those in HPDE cells (0.95±0.02, 1.13±0.03, t =25.940, 24.500, P＜0.01). At 72 and 96 hours, the cell proliferation ability and number of clones cell in the miR-21-minic group were significantly higher than those in the miR-NC group, while the number of apoptotic cells was lower than that in the miR-NC group (P＜0.05). The glucose consumption, and the content of lactate, hexokinase (HK) and lactate dehydrogenase (LDH) in miR-21-mimic group were significantly higher than those in miR-NC group ( t =5.992, 21.340, 5.643, 4.008, P＜0.05). The level of PKM2 protein in the miR-21 mic group was lower than that in the miR-NC group ( t =10.070, P＜0.01). There were a certain number of complementary base pairs between miR-21 and WT-3′ UTR-PKM2, and the cell luciferase activity of miR-21-minic+WT-PKM2 group was lower than that of the transfected miR-NC+WT-PKM2 group ( t =15.680, P＜0.01). Conclusion The overexpression of miR-21 in pancreatic cancer cells may promote the proliferation, aerobic glycolysis, and inhibit apoptosis of pancreatic cancer cells. The mechanism may be related to targeting PKM2.
文章编号： 中图分类号：R735.9 文献标志码：A
基金项目:新疆维吾尔自治区自然科学基金（2021D01C375）
附件
| Author Name | Affiliation |
| LI Jiangang, Paheredini Yusupu, WANG Jun, LI Liang | General Surgery Department, The Second Affiliated Hospital of Xinjiang Medical University, Urumqi, Xinjiang 830063, China |
引用文本：
李建刚，帕合热迪尼·玉素莆，王俊，李亮.微小RNA-21过表达对胰腺癌细胞增殖和凋亡及有氧糖酵解的影响[J].中国临床研究,2023,3(9):1286-1290.
李建刚，帕合热迪尼·玉素莆，王俊，李亮.微小RNA-21过表达对胰腺癌细胞增殖和凋亡及有氧糖酵解的影响[J].中国临床研究,2023,3(9):1286-1290.