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].,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 |
投稿时间:2023-05-29 修订日期:2023-06-25 |
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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