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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

27卷

期数:

2021年04期

页码:

343-348,378

栏目:

基础医学

出版日期:

2021-07-20

文章信息/Info

Title:

The effect of NAMPT on the proliferation and apoptosis of renal cell carcinoma and its mechanism

文章编号:

1006－8147（2021）04-0343-07

作者:

王艳辉; 柳雅慧; 任丽

（天津医科大学肿瘤医院检验科，国家肿瘤临床医学研究中心，天津市“肿瘤防治”重点实验室，天津市恶性肿瘤临床医学研究中心，天津300060）

Author(s):

WANG Yan-hui; LIU Ya-hui; REN Li

（Department of Laboratory，Tianjin Medical University Cancer Institute and Hospital，National Clinical Research Center for Cancer，Tianjin Key Laboratory of Cancer Prevention and Therapy，Tianjin Clinical Research Center for Cancer，Tianjin 300060，China）

关键词:

肾癌; NAMPT; 增殖; 细胞凋亡

Keywords:

renal cancer; NAMPT; proliferation; apoptosis

分类号:

R737.11

DOI:

-

文献标志码:

A

摘要:

目的：探讨烟酰胺磷酸核糖转移酶(NAMPT) 在肾癌中的表达情况及对肾癌细胞增殖和凋亡的影响。方法：通过TCGA在线数据库ualcan分析NAMPT在肾癌组织和正常肾组织中的表达，通过Kaplan-Meier plotter数据库分析NAMPT对肾癌预后的影响。qPCR检测NAMPT基因在肾癌细胞769P、A704、786-O及正常化肾小管上皮细胞HK2中的表达。分别设计NAMPT特异性shRNA（shNAMPT组）和对照序列（scramble组）转染肾癌细胞786-O，用NAMPT抑制剂FK866及对照试剂DMSO处理肾癌细胞786-O，分别采用CCK-8法、平板克隆实验及流式细胞术检测NAMPT对肾癌细胞786-O增殖、克隆形成及凋亡的影响，NAD+/NADH检测试剂盒检测细胞内烟酰胺腺嘌呤二核苷酸（NAD+）的含量，Western印迹检测NAMPT的表达及干扰NAMPT对磷脂酰肌醇3激酶/蛋白激酶B（PI3K/Akt）通路的影响。结果：生物信息学分析显示NAMPT在肾癌中的表达高于正常肾组织（P＜0.05），NAMPT高表达组总生存率较低表达组明显降低（P<0.01）；肾癌细胞769P、A704、786-ONAMPT的表达均高于肾小管上皮细胞HK2（t=6.680，P=0.0026，t=14.12，P=0.001，t=13.43，P=0.002）；细胞学实验结果显示，与DMSO组相比，FK866组肾癌细胞786-O的增殖能力减弱（t=2.625，P<0.05），克隆形成能力明显受抑制（t=5.687，P<0.01），凋亡细胞比例增高（t=6.325，P＜0.01），NAD+生成明显下降（t=17.62，P<0.001）。与scramble组相比，shNAMPT组肾癌细胞786-O的增殖能力减弱（t=3.959，P<0.05），克隆形成能力明显受抑制（t=8.684，P<0.05），凋亡细胞比例增高（t=14.05，P＜0.01），NAD+生成明显下降（t=10.93，P<0.001），Akt通路关键蛋白p-PI3K及p-Akt的表达明显下调（t=7.721，P＜0.01，t=14.78，P＜0.001）。结论：NAMPT在肾癌中高表达，且与患者预后差相关，干扰NAMPT的表达，可以通过减少NAD+的生成，抑制PI3K/Akt通路的激活，从而降低肾癌细胞的增殖，促进其凋亡。

Abstract:

Objective: To investigate the expression of nicotinamide phosphoribosyl transferase（NAMPT） in renal cell carcinoma and its effect on the proliferation and apoptosis of renal cell carcinoma. Methods: The TCGA online database ualcan was used to analyze the expression of NAMPT in renal cancer tissues and normal renal tissues，the Kaplan-Meier plotter database was used to analyze the influence of NAMPT on the prognosis of renal cancer. qPCR was used to detect the expression of NAMPT gene in renal cancer cell 769P，A704，786-O and normalized renal tubular epithelial cells HK 2. 786-O cell was transfected with NAMPT specific shRNA（shNAMPT group） and control sequence（scramble group） respectively，and treated with NAMPT inhibitor FK866 and control reagent DMSO. CCK-8 assay，cell colony formation assay and flow cytometry were respectively used to detect cell proliferation，the cloning ability and apoptosis of 786-O cell. NAD+/NADH detection kit was used to detect the content of nicotinamide adenine dinucleotide（NAD+） in cells. Western blotting assay was used to detect the expression of NAMPT and the effect of interfering with NAMPT on phosphatidylinositol 3 kinase/protein kinase B （PI3K / Akt） pathway. Results: Bioinformatics analysis showed that the expression of NAMPT in renal cell carcinoma was higher than that in normal renal tissues（P＜0.05）. The overall survival rate was significantly reduced in patients with high expression of NAMPT group than those in patients with low expression of NAMPT group（P＜0.01）；the expression of NAMPT were higher in 769P，A704，786-O than those in renal tubular epithelial cells HK2（t=6.680，P=0.0026，t=14.12，P=0.001，t=13.43，P=0.002）.Cytological experiment results showed that compared with DMSO group，in FK866 group，786-O cell proliferation and clone formation ability was significantly inhibited（t=2.625，P<0.05，t=5.687，P<0.01），and the proportion of apoptotic cells was increased（t=6.325，P＜0.01），the production of NAD+ was significantly decreased （t=17.62，P<0.001）.Compared with scramble group，in shNAMPT group，786-O cell proliferation and clone formation ability was significantly inhibited（t=3.959，P<0.05，t=8.684，P<0.05），and the proportion of apoptotic cells was increased（t=14.05，P＜0.01），the production of NAD+ was significantly decreased（t=10.93，P<0.001），and the expression of key proteins p-PI3K and p-Akt of the Akt pathway were significantly down-regulated（t=7.721，P＜0.01，t=14.78，P＜0.001）. Conclusion: NAMPT is highly expressed in renal cell carcinoma and is related to the poor prognosis of patients. Interfering with the expression of NAMPT can reduce the production of NAD+ and inhibit the activation of PI3K/Akt pathway，thereby reducing the proliferation and promoting apoptosis of renal cancer cells.

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[2]任 静,朱仲玲,阎 昭.肾癌舒尼替尼耐药细胞株ACSUR的建立及其生物学特性研究[J].天津医科大学学报,2019,25(06):556.
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备注/Memo

备注/Memo:

作者简介：王艳辉（1992-），女，硕士在读，研究方向：肿瘤微环境和肿瘤代谢；通信作者：任丽，E-mail：roland-li@163.com。

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更新日期/Last Update: 2021-07-25