长链非编码RNA AFAP1-AS1通过PTEN/p-AKT信号通路调控结直肠癌细胞增殖的分子机制研究

doi:10.11904/j.issn.1002-3070.2020.01.003

实用肿瘤学杂志 ›› 2020, Vol. 34 ›› Issue (1): 11-16.doi: 10.11904/j.issn.1002-3070.2020.01.003

长链非编码RNA AFAP1-AS1通过PTEN/p-AKT信号通路调控结直肠癌细胞增殖的分子机制研究

曾理¹, 庄树彤², 丁世华¹, 陈冲¹, 焦璐¹, 孙大勇¹

1.深圳大学第一附属医院(深圳市第二人民医院)消化内科(深圳 518035);
2.深圳大学第一附属医院(深圳市第二人民医院)胃肠外科

收稿日期:2019-08-27 修回日期:2019-11-13 出版日期:2020-02-28 发布日期:2020-02-20
通讯作者: 孙大勇,E-mail:chndoctor@21cn.com
作者简介:曾理,男,(1979-),博士,副主任医师,从事消化道肿瘤的早期诊断及治疗的研究。
基金资助:
深圳市卫生计生系统科研项目(编号:201601021;201601024)

Molecular mechanisms of long non-coding RNA AFAP1-AS1 regulating the proliferation of colorectal cancer cells by PTEN/p-AKT signaling pathway

ZENG Li¹, ZHUANG Shutong², DING Shihua¹, CHEN Chong¹, JIAO Lu¹, SUN Dayong¹

1.Department of Gastroenterology,The First Affiliated Hospital of Shenzhen University,The Second People′s Hospital of Shenzhen,Shenzhen,518035,China;
2.Department of Gastrointestinal Surgery,The First Affiliated Hospital of Shenzhen University,The Second People′s Hospital of Shenzhen

Received:2019-08-27 Revised:2019-11-13 Online:2020-02-28 Published:2020-02-20

摘要/Abstract

摘要： 目的探讨长链非编码RNA(lncRNA)肌动蛋白纤维相关蛋白1-反义RNA1(Actin filament-associatedprotein1-antisenseRNA1,AFAP1-AS1)调控结直肠癌(Colorectal cancer,CRC)细胞增殖的分子机制。方法收集2014年7月—2018年6月深圳市第二人民医院消化内科和胃肠外科诊治的38例正常人及38例CRC患者的粪便标本,用Real time-PCR检测lncRNA AFAP1-AS1表达,同时检测人正常结肠上皮细胞株NCM460、人结直肠癌细胞株SW620和HCT116中lncRNA AFAP1-AS1的表达;将靶向siRNA转染到CRC细胞株SW620和HCT116中抑制AFAP1-AS1的表达;通过MTT测定si-AFAP1-AS1转染对CRC细胞增殖的影响;应用免疫印迹检测Cleaved caspase 3、Bcl-2、Bax、p-AKT、total-AKT和PTEN的水平。结果与正常人相比,CRC患者粪便中AFAP1-AS1的表达显著上升(P＜0.01);与NCM460细胞相比,SW620和HCT116细胞中AFAP1-AS1的表达也显著升高(P＜0.01);si-AFAP1-AS1转染抑制SW620和HCT116的细胞生长(P＜0.01)。与NCM460细胞相比,si-AFAP1-AS1干扰上调了SW620和HCT116细胞中Cleaved caspase 3和促凋亡蛋白Bax的表达水平(P＜0.05),下调了抗凋亡蛋白Bcl-2的表达水平(P＜0.001);此外,si-AFAP1-AS1干扰降低了CRC细胞中p-AKT的蛋白水平,并增加了PTEN的表达(P＜0.01)。结论正常人和CRC患者粪便中lncRNA AFAP1-AS1表达的差异有助于CRC的早期诊断,且lncRNA AFAP1-AS1在CRC细胞中表达上调,并通过PTEN/p-AKT信号通路调节CRC细胞的增殖和凋亡。lncRNA AFAP1-AS1有望成为CRC早期诊断的分子靶标。

关键词: AFAP1-AS1, 结直肠癌, 凋亡, PTEN/p-AKT信号通路

Abstract: Objective The aim of this study was to explore the molecular mechanism of long non-coding RNA(LncRNA)AFAP1-AS1 regulating the proliferation of colorectal cancer(CRC)cells.Methods Fecal specimens from 38 normal people and 38 CRC patients diagnosed and treated in the department of digestive medicine and gastrointestinal surgery of Shenzhen Second People′s Hospital from July 2014 to June 2018 were collected to detect the expression of lncRNA AFAP1-AS1 by Real time-PCR.At the same time,the expression of lncRNA AFAP1-AS1 was detected in human normal colon epithelial NCM460 cells,human colorectal cancer SW620 and HCT116 cells.Targeting lncRNA AFAP1-AS1 siRNA(si-AFAP1-AS1)was transfected into SW620 and HCT116 cells to inhibit the expression of AFAP1-AS1.MTT assay was used to examine the effect of si-AFAP1-AS1 on cell proliferation of CRC cells.The levels of cleaved-caspase-3,Bcl-2,Bax,p-AKT,total AKT,and PTEN were detected by Western blot.Results Compared with the normal control group,the expression of AFAP1-AS1 in feces of CRC patients significantly increased(P＜0.01);Compared with NCM460 cells,the expression of AFAP1-AS1 in SW620 and HCT116 cells also significantly increased(P＜0.01);si-AFAP1-AS1 significantly suppressed the cell viability of SW620 and HCT116 cells(P＜0.01).Compared with NCM460 cells,si-AFAP1-AS1 up-regulated the levels of cleaved-caspase-3 and pro-apoptotic protein Bax(P＜0.05),and down-regulated the level of anti-apoptotic protein Bcl-2 in SW620 and HCT116 cells(P＜0.001).In addition,si-AFAP1-AS1 significantly decreased the level of p-AKT protein and increased the expression of PTEN in CRC cells(P＜0.01).Conclusion The difference in the expression of lncRNA AFAP1-AS1 in the stool of normal and CRC patients is beneficial to the early diagnosis for CRC,the expression of lncRNA AFAP1-AS1 is up-regulated in CRC cells regulated by the PTEN/p-AKT signaling pathway.Therefore,lncRNA AFAP1-AS1 is expected to become a molecular target for early diagnosis of colorectal cancer.

Key words: AFAP1-AS1, Colorectal cancer, Apoptosis, PTEN/p-AKT signaling pathway

中图分类号:

R735.34

曾理, 庄树彤, 丁世华, 陈冲, 焦璐, 孙大勇. 长链非编码RNA AFAP1-AS1通过PTEN/p-AKT信号通路调控结直肠癌细胞增殖的分子机制研究[J]. 实用肿瘤学杂志, 2020, 34(1): 11-16.

ZENG Li, ZHUANG Shutong, DING Shihua, CHEN Chong, JIAO Lu, SUN Dayong. Molecular mechanisms of long non-coding RNA AFAP1-AS1 regulating the proliferation of colorectal cancer cells by PTEN/p-AKT signaling pathway[J]. Journal of Practical Oncology, 2020, 34(1): 11-16.

参考文献

1 Shike M,Winawer SJ,Greenwald PH,et al.Primary prevention of colorectal cancer.The WHO Collaborating Centre for the Prevention of Colorectal Cancer[J].B World Health Organ,1990,68(3):377-385.
2 Svoboda M,Slyskova J,Schneiderova M,et al.HOTAIR long non-coding RNA is a negative prognostic factor not only in primary tumors,but also in the blood of colorectal cancer patients[J].Carcinogenesis,2014,35(7):1510-1515.
3 夏天一,张博淼,姚克非,等.PTPN9表达水平与结直肠癌的临床关系研究[J].实用肿瘤学杂志,2019,33(5):415-419.
4 Lecerf C,Le Bourhis X,Adriaenssens E.The long non-coding RNA H19:an active player with multiple facets to sustain the hallmarks of cancer[J].Cell Mol Life Sci,2019[Epub ahead of print].
5 Ji D,Zhong X,Jiang X,et al.The role of long non-coding RNA AFAP1-AS1 in human malignant tumors[J].Pathol Res and Pract,2018,214(10):1524-1531.
6 Luo HL,Huang MD,Guo JN,et al.AFAP1-AS1 is upregulated and promotes esophageal squamous cell carcinoma cell proliferation and inhibits cell apoptosis[J].Cancer Med,2016,5(10):2879-2885.
7 Han X,Wang L,Ning Y,et al.Long non-coding RNA AFAP1-AS1 facilitates tumor growth and promotes metastasis in colorectal cancer[J].Biol Res,2016,49(1):36.
8 Wang F,Ni H,Sun F,et al.Overexpression of lncRNA AFAP1-AS1 correlates with poor prognosis and promotes tumorigenesis in colorectal cancer[J].Biomed Pharmacother,2016,81:152-159.
9 Conley-LaComb MK,Saliganan A,Kandagatla P,et al.PTEN loss mediated Akt activation promotes prostate tumor growth and metastasis via CXCL12/CXCR4 signaling[J].Mol Cancer,2013,12(1):85.
10 Xiao ZD,Jiao CY,Huang HT,et al.miR-218 modulate hepatocellular carcinoma cell proliferation through PTEN/AKT/PI3K pathway and HoxA10[J].Int J Clin Exp Patho,2014,7(7):4039-4044.
11 Gao C,Yuan X,Jiang Z,et al.Regulation of AKT phosphorylation by GSK3beta and PTEN to contr FEBS Journal ol chemoresistance in breast cancer[J].Breast Cancer Res Treat,2019,176(2):291-301.
12 Badr G,Sayed EA,Waly H,et al.The therapeutic mechanisms of propolis against CCl4-mediated liver injury by mediating apoptosis of activated hepatic stellate cells and improving the hepatic architecture through PI3K/AKT/mTOR,TGF-beta/Smad2,Bcl2/BAX/P53 and iNOS signaling pathways[J].Cell Physiol Biochem,2019,53(2):301-322.
13 Larsen BD,Sorensen CS.The caspase-activated DNase:apoptosis and beyond[J].FEBS J,2017,284(8):1160-1170.

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长链非编码RNA AFAP1-AS1通过PTEN/p-AKT信号通路调控结直肠癌细胞增殖的分子机制研究

Molecular mechanisms of long non-coding RNA AFAP1-AS1 regulating the proliferation of colorectal cancer cells by PTEN/p-AKT signaling pathway

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