| 孙良璋,徐正水,张丹杰,马震川,李建忠,孔冉冉,张 晋.Nampt及高糖高胰岛素微环境对人顺铂耐药肺癌细胞株增殖和转移的影响[J].现代生物医学进展英文版,2023,(24):4620-4626. |
| Nampt及高糖高胰岛素微环境对人顺铂耐药肺癌细胞株增殖和转移的影响 |
| Effects of Nampt and High Glucose and High Insulin Microenvironment on Proliferation and Metastasis of Human Cisplatin-resistant Lung Cancer Cell Lines |
| Received:May 29, 2023 Revised:June 25, 2023 |
| DOI：10.13241/j.cnki.pmb.2023.24.004 |
| 中文关键词: 烟酰胺磷酸核糖转移酶 高糖 高胰岛素 肺癌 顺铂 |
| 英文关键词: Nicotinamide phosphoribosyltransferase High glucose High insulin Lung cancer Cisplatin |
| 基金项目:陕西省自然科学基础研究计划项目（2023-JC-QN-0840）；西安市科技计划项目（2019114613YX001SF034(7)） |
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| 中文摘要: |
| 摘要 目的：探究烟酰胺磷酸核糖转移酶（Nampt）及高糖高胰岛素（HG+HI）微环境对人顺铂耐药肺癌细胞增殖和转移的影响。方法：将人顺铂耐药细胞株A549/DDP分为6组（n=6）：对照组（control）、高糖高胰岛素干预组（HH，使用添加30 mmol/L的葡萄糖和500 mU/L的胰岛素的培养基培养72 h）、分别转染sh-NC和sh-Nampt组（sh-NC和sh-Nampt，使用Lipofectamine 2000将sh-NC和sh-Nampt分别转染到细胞中，转染时间为48 h）、HH干预sh-NC和sh-Nampt组（HH+sh-NC和HH+sh-Nampt）。每组6个重复样本。qRT-PCR检测转染效率，MTT法检测细胞增殖，流式细胞仪检测细胞凋亡，Transwell检测细胞迁移和侵袭，qRT-PCR检测Nampt mRNA，Western blot检测Nampt、Bcl-2、Bax、MMP-2、MMP-9、p-PI3K、PI3K、p-AKT和AKT蛋白表达。结果：与对照组和sh-NC组比较，sh-Nampt组的Nampt mRNA和蛋白表达水平、相对细胞活力、迁移和侵袭细胞数量降低，而细胞凋亡率升高，Bcl-2、MMP-2、MMP-9、Nampt、p-PI3K和p-AKT蛋白表达水平降低，Bax蛋白表达水平升高（P<0.005）。与对照组和sh-NC组比较，HH组和HH+sh-NC组的Nampt mRNA和蛋白表达水平、相对细胞活力、迁移和侵袭细胞数量升高，Bcl-2、MMP-2、MMP-9、Nampt、p-PI3K和p-AKT蛋白表达水平升高，Bax蛋白表达水平降低（P<0.005）。与HH组和HH+sh-NC组比较，HH+sh-Nampt组的Nampt mRNA和蛋白表达水平、相对细胞活力、迁移和侵袭细胞数量降低，细胞凋亡率升高，Bcl-2、MMP-2、MMP-9、Nampt、p-PI3K和p-AKT蛋白表达水平降低，Bax蛋白表达水平升高（P<0.005）。结论：高糖高胰岛素微环境可能通过上调Nampt/PI3K/AKT信号通路诱导人顺铂耐药肺癌细胞的增殖和转移。 |
| 英文摘要: |
| ABSTRACT Objective: To reveal the effects of nicotinamide phosphoribosyltransferase (Nampt) and high glucose and high insulin (HG+HI) microenvironment on proliferation and metastasis of human cisplatin-resistant lung cancer cell lines. Methods: Human cisplatin resistant cell line A549/DDP was divided into 6 groups (n=6): control group, high glucose and high insulin intervention group (HH, cultured with 30 mmol/L glucose and 500 mU/L insulin for 72 h), sh-NC and sh-Nampt transfected groups (Lipofectamine 2000 was used to transfect sh-NC and sh-Nampt into cells respectively, the transfection time was 48 h), HH intervention sh-NC and sh-Nampt groups (HH+sh-NC and HH+sh-Nampt). Each group had 6 replicates. The transfection efficiency was detected by qRT-PCR. Cell proliferation was detected by MTT method, apoptosis was detected by flow cytometry, cell migration and invasion were detected by Transwell, Nampt mRNA was detected by qRT-PCR, and protein expression of Nampt, Bcl-2, Bax, MMP-2, MMP-9, p-PI3K, PI3K, p-AKT and AKT were detected by Western blot. Results: Compared with control group and sh-NC group, the expression level of Nampt mRNA and protein, relative cell viability, the number of migratory and invasive cells decreased, the rate of apoptosis increased, the expression level of Bcl-2, MMP-2, MMP-9, Nampt, p-PI3K and p-AKT protein decreased, and the expression level of Bax protein increased in sh-Nampt group (P<0.05). Compared with control group and sh-NC group, the expression level of Nampt mRNA and protein, relative cell viability, the number of migratory and invasive cells increased, the expression level of Bcl-2, MMP-2, MMP-9, Nampt, p-PI3K and p-AKT protein increased, and the expression level of Bax protein decreased in HH group and HH+sh-NC group(P<0.05). Compared with HH group and HH+sh-NC group, the expression level of Nampt mRNA and protein, relative cell viability, the number of migratory and invasive cells decreased, the rate of apoptosis increased, the expression level of Bcl-2, MMP-2, MMP-9, Nampt, p-PI3K and p-AKT protein decreased, and the expression level of Bax protein increased in HH+sh-Nampt group(P<0.05). Conclusion: High glucose and high insulin microenvironment may induce proliferation and metastasis of human cisplatin-resistant lung cancer cells by up-regulating Nampt/PI3K/AKT signaling pathway. |
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