AKR1B10基因沉默对肝癌细胞增殖和凋亡的影响

钱瑞坤; 马长林; 乔森

doi:10.3969/j.issn.1001-5256.2020.04.018

AKR1B10基因沉默对肝癌细胞增殖和凋亡的影响

DOI: 10.3969/j.issn.1001-5256.2020.04.018

济宁市第一人民医院肝胆外科

基金项目:

济宁市科技助推新旧动能转换计划(2018SMN007)；

详细信息

中图分类号: R735.7
计量
- 文章访问数: 1017
- HTML全文浏览量: 14
- PDF下载量: 210
- 被引次数: 0
出版历程
- 收稿日期: 2019-11-06
- 出版日期: 2020-04-20

Effect of Aldo-keto reductase family 1 member B10 gene silencing on the proliferation and apoptosis of hepatocellular carcinoma cells

Department of Hepatobiliary Surgery,The First People's Hospital of Jining

Research funding:

摘要

摘要: 目的探索肝细胞癌（HCC）中醛酮还原酶家族1成员B10（AKR1B10）的生物学功能和相关的病理机制。方法通过CCLE数据库分析正常肝细胞与HCC细胞系中AKR1B10 mRNA的表达,UALCAN以及Oncomine数据库分析癌旁组织与HCC组织中AKR1B10 mRNA的表达,Western Blot法验证HCC患者癌组织与癌旁组织中AKR1B10蛋白表达差异。使用慢病毒分别敲减HepG2及Huh7细胞中的AKR1B10,分为对照组和AKR1B10敲减组（shAKR1B10 1#、shAKR1B10 2#）。采用AnnexinV-APC/PI双染法、细胞克隆实验、皮下移植肿瘤的方法检测敲减AKR1B10后HCC细胞凋亡、克隆形成、皮下成瘤体积的变化。过氧化物敏感性荧光探针DCFH-DA以及Western Blot检测敲减AKR1B10后HCC细胞内活性氧（ROS）以及p-ATMser1981、γ-H2AX、c-Caspase-3和c-RARP等蛋白的变化。计量资料多组间比较采用单因素方差分析,进一步两两比较采用SNK-q检验。结果 CCLE数据库提示AKR1B...
- 肝肿瘤,实验性 /
- 醛酮还原酶家族1成员B10 /
- 基因沉默 /
- 活性氧
Abstract: Objective To investigate the biological functions and related pathological mechanisms of Aldo-keto reductase family 1 member B10(AKR1 B10) in hepatocellular carcinoma(HCC).Methods The COLE database was used to analyze the mRNA expression of AKR1 B10 in normal liver cells and HCC cell lines.The UALCAN and Oncomine databases were used to analyze the mRNA expression of AKR1 B10 in HCC tissue and adjacent tissue.Western blot was used to investigate the difference in the protein expression of AKR1 B10 between HCC tissue and adjacent tissue.AKR1 B10 in HepG2 and Huh7 cells was knocked down by lentivirus,and the cells were divided into control group and AKR1 B10 knockdown group(which was further divided into shAKR1 B10 1# group and shAKR1 B10 2# group).AnnexinVAPC/PI double staining,colony formation assay,and the method of subcutaneous xenograft tumor were used to observe the apoptosis of HCC cells,colony formation,and the change in the volume of subcutaneous xenograft tumor after AKR1 B10 knockdown.The peroxide-sensitive fluorescent probe DCFH-DA and Western blot were used to measure the changes in ROS and related proteins,such as pATMser1981,γ-H2 AX,c-Caspase-3,and c-RARP in HCC cells.A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the SNK-q test was used for further comparison between two groups.Results The CCLE database showed that the mRNA expression of AKR1 B10 in HCC cells was significantly higher than that in normal liver cells) the Oncomine database suggested that the mRNA expression of AKR1 B10 in HCC tissue was more than 10 times that in adjacent tissue) the UALCAN database showed that the AKR1 B10 gene was highly expressed in HCC tissue.For the six patients in this study,the expression of AKR1 B10 in HCC tissue was significantly higher than that in adjacent tissue.Compared with the control group,the AKR1 B10 knockdown group had a significant reduction in the colony formation ability of HCC cells(P <0.001) and a significant increase in apoptosis rate(P <0.001).Compared with the control group,the AKR1 B10 knockdown group had significant increases in ROS,the markers for DNA damage p-ATMser1981 and γ-H2 AX,and the indices for cell apoptosis c-Caspase-3 and c-RARP.In the BALB-c/Nude mouse model,compared with the control group,the AKR1 B10 knockdown group had a significant reduction in tumor volume since day 13 after allograft transplantation(P <0.05).Conclusion AKR1 B10 may be an effective therapeutic target for the treatment of HCC.
- liver neoplasms,experimental /
- AKR1B10 /
- gene silencing /
- reactive oxygen species

HTML全文

参考文献(14)

[1] MA J,LUO DX,HUANG C,et al. AKR1B10 overexpression in breast cancer:Association with tumor size,lymph node metastasis and patient survival and its potential as a novel serum marker[J]. Int J Cancer,2012,131(6):e862-e871.

[2] SATO S,GENDA T,ICHIDA T,et al. Impact of aldo-keto reductase family 1 member B10 on the risk of hepatitis C virusrelated hepatocellular carcinoma[J]. J Gastroenterol Hepatol,2016,31(7):1315-1322.

[3] CHUNG YT,MATKOWSKYJ KA,LI H,et al. Overexpression and oncogenic function of aldo-keto reductase family 1B10(AKR1B10)in pancreatic carcinoma[J]. Mod Pathol,2012,25(5):758-766.

[4] FUKUMOTO S,YAMAUCHI N,MORIGUCHI H,et al. Overexpression of the aldo-keto reductase family protein AKR1B10is highly correlated with smokers’non-small cell lung carcinomas[J]. Clin Cancer Res,2005,11(5):1776-1785.

[5] CHEN WD,ZHANG Y. Regulation of aldo-keto reductases in human diseases[J]. Front Pharmacol,2012,3:35.

[6] GIANNINI EG,MARENCO S,BRUZZONE L,et al. Hepatocellular carcinoma in patients without cirrhosis in Italy[J]. Dig Liver Dis,2013,45(2):164-169.

[7] KULIK LM,CHOKECHANACHAISAKUL A. Evaluation and management of hepatocellular carcinoma[J]. Clin Liver Dis,2015,19(1):23-43.

[8] GRANJA S,PINHEIRO C,REIS RM,et al. Glucose addiction in cancer therapy:Advances and drawbacks[J]. Curr Drug Metab,2015,16(3):221-242.

[9] GANAPATHY-KANNIAPPAN S,GESCHWIND JF. Tumor glycolysis as a target for cancer therapy:Progress and prospects[J]. Mol Cancer,2013,12:152.

[10] PANIERI E,SANTORO MM. ROS homeostasis and metabolism:A dangerous liason in cancer cells[J]. Cell Death Dis,2016,7(6):e2253.

[11] LEE JH,PAULL TT. Activation and regulation of ATM kinase activity in response to DNA double-strand breaks[J]. Oncogene,2007,26(56):7741-7748.

[12] REDON CE,DICKEY JS,BONNER WM,et al．γ-H2AX as a biomarker of DNA damage induced by ionizing radiation in human peripheral blood lymphocytes and artificial skin[J]. Adv Space Res,2009,43(8):1171-1178.

[13] TONG L,CHUANG CC,WU S,et al. Reactive oxygen species in redox cancer therapy[J]. Cancer Lett,2015,367(1):18-25.

[14] WANG Z,LI S,CAO Y,et al. Oxidative stress and carbonyl lesions in ulcerative colitis and associated colorectal cancer[J]. Oxid Med Cell Longev,2016,2016:9875298.

施引文献

资源附件(0)

访问统计