长链非编码RNA CASC15对肝细胞癌增殖、侵袭和迁移的影响及作用机制

doi:10.11904/j.issn.1002-3070.2024.01.004

实用肿瘤学杂志 ›› 2024, Vol. 38 ›› Issue (1): 18-29.doi: 10.11904/j.issn.1002-3070.2024.01.004

长链非编码RNA CASC15对肝细胞癌增殖、侵袭和迁移的影响及作用机制

吴东洋, 蔡青山, 刘东, 左刚刚, 李树栋, 刘立友, 郑建兴

唐山中心医院肝胆外科(唐山 063000)

收稿日期:2023-09-12 修回日期:2024-01-29 出版日期:2024-02-28 发布日期:2024-06-21
通讯作者: 郑建兴,E-mail:ZJX_19740@163.com
作者简介:吴东洋,男,(1986-),硕士,主治医师,从事肝胆肿瘤方向的研究。
基金资助:
河北省医学科学研究课题(编号:20221840)

The effects and mechanism of long non-coding RNA CASC15 on proliferation,invasion and migration of hepatocellular carcinoma

WU Dongyang, CAI Qingshan, LIU Dong, ZUO Ganggang, LI Shudong, LIU Liyou, ZHENG Jianxing

Department of Hepatobiliary Surgery,Tangshan Central Hospital,Tangshan 063000,China

Received:2023-09-12 Revised:2024-01-29 Online:2024-02-28 Published:2024-06-21

摘要/Abstract

摘要： 目的探究长链非编码RNA(lncRNA)癌症易感性基因15(Cancer susceptibility candidate 15,CASC15)在肝细胞癌(Hepatocellular carcinoma,HCC)中的分子调控机制。方法通过生物信息学方法预测目的基因表达,并分析目的基因表达与患者生存时间的关系;收集临床HCC患者肝癌组织和癌旁组织;CCK-8、Transwell和流式细胞术实验检测HCC细胞SMMC7721和Huh-7的增殖、侵袭、迁移以及凋亡;双荧光素酶实验检测miR-144-3p与CASC15和富含亮氨酸的重复序列蛋白1(Leucine rich repeat containing protein 1,LRRC1)的靶向关系;qRT-PCR和Western blot检测mRNA和蛋白表达情况;免疫荧光实验用于蛋白定位研究;回复实验验证CASC15/miR-144-3p/LRRC1对HCC进展的影响。体内实验验证CASC15对HCC进展的影响。结果 TCGA数据库与qRT-PCR检测显示HCC组织和细胞中CASC15高表达、miR-144-3p低表达、LRRC1高表达(P＜0.05)。增殖、侵袭、迁移的细胞功能实验结果表明在肿瘤发展中CASC15和LRRC1起到促进作用,miR-144-3p则是抑制作用,与凋亡实验结果一致(P＜0.05)。细胞功能实验表明CASC15抑制miR-144-3p功能,miR-144-3p抑制LRRC1,CASC15与miR-144-3p结合,并导致LRRC1上调。回复实验结果表明CASC15通过抑制miR-144-3p促进LRRC1的表达从而促进HCC细胞增殖、侵袭和迁移并抑制细胞凋亡。结论 CASC15可能通过调节miR-144-3p/LRRC1轴,从而促进HCC进展。

关键词: 癌症易感性基因15, miR-144-3p, 富含亮氨酸的重复序列蛋白1, 肝癌细胞, 细胞生物学功能

Abstract: Objective The aim of this study was to investigate the molecular regulatory mechanism of cancer susceptibility candidate 15 (CASC15),a long-stranded non-coding RNA (lncRNA),in hepatocellular carcinoma (HCC). Methods Bioinformatics methods were used to predict the expression of target genes and analyze the relationship between the expression of target genes and the survival time of patients;Hepatocellular carcinoma tissues and adjacent tissues from patients with HCC were collected;CCK-8,Transwell,and flow cytometry experiments were used to detect proliferation,invasion,migration and apoptosis of SMMC7721 cells and Huh-7 cells;The dual-luciferase assay was used to detect the targeting relationship between miR-144-3p and CASC15,as well as leucine rich repeat containing protein 1 (LRRC1);RT-qPCR and Western blot were used to detect mRNA and protein expression of target genes;Immunofluorescence was used for protein localization of target genes;Replicate experiment was performed to verify the effect of CASC15/miR-144-3p/LRRC1 on the progression of HCC.In vivo experiment was performed to verify the effect of CASC15 on HCC progression. Results TCGA database and RT-qPCR assay showed high expression of CASC15,low expression of miR-144-3p,and high expression of LRRC1 in HCC tissues and cells (P＜0.05).The results of cell function experiments on proliferation,invasion and migration showed that CASC15 and LRRC1 played a promoting role in tumor development,while miR-144-3p had an inhibitory effect,consistent with the results of apoptosis experiments (P＜0.05).Cell function experiments showed that CASC15 inhibited miR-144-3p function,miR-144-3p inhibited LRRC1,and CASC15 bound to miR-144-3p,leading to the upregulation of LRRC1.The replicate experimental results indicated that CASC15 promoted LRRC1 expression through inhibiting miR-144-3p,thereby promoting HCC cell proliferation,invasion and migration,and inhibiting apoptosis. Conclusion CASC15 may promote HCC progression by regulating the miR-144-3p/LRRC1 axis.

Key words: Cancer susceptibility gene 15, miR-144-3p, Leucine-rich repeative sequence protein 1, Hepatocellular carcinoma cells, Cell biological function

中图分类号:

R735.7

吴东洋, 蔡青山, 刘东, 左刚刚, 李树栋, 刘立友, 郑建兴. 长链非编码RNA CASC15对肝细胞癌增殖、侵袭和迁移的影响及作用机制[J]. 实用肿瘤学杂志, 2024, 38(1): 18-29.

WU Dongyang, CAI Qingshan, LIU Dong, ZUO Ganggang, LI Shudong, LIU Liyou, ZHENG Jianxing. The effects and mechanism of long non-coding RNA CASC15 on proliferation,invasion and migration of hepatocellular carcinoma[J]. Journal of Practical Oncology, 2024, 38(1): 18-29.

参考文献

1 Longerich T.Hepatocellular carcinoma[J].Pathologe,2020,41(5):478-487.
2 Rimassa L,Pressiani T,Merle P.Systemic treatment options in hepatocellular carcinoma[J].Liver Cancer,2019,8(6):427-446.
3 Couri T,Pillai A.Goals and targets for personalized therapy for HCC[J].Hepatol Int,2019,13(2):125-137.
4 Hajjari M,Salavaty A.HOTAIR:an oncogenic long non-coding RNA in different cancers[J].Cancer Biol Med,2015,12(1):1-9.
5 Liu J,Peng WX,Mo YY,et al.MALAT1-mediated tumorigenesis[J].Front Biosci (Landmark Ed),2017,22(1):66-80.
6 Asghar U,Witkiewicz AK,Turner NC,et al.The history and future of targeting cyclin-dependent kinases in cancer therapy[J].Nat Rev Drug Discov,2015,14(2):130-146.
7 Su C,Wang W,Mo J,et al.Long noncoding RNA HOXC-AS3 interacts with CDK2 to promote proliferation in hepatocellular carcinoma[J].Biomark Res,2022,10(1):65.
8 Chandra Gupta S,Nandan Tripathi Y.Potential of long non-coding RNAs in cancer patients:From biomarkers to therapeutic targets[J].Int J Cancer,2017,140(9):1955-1967.
9 Zhang DY,Sun QC,Zou XJ,et al.Long noncoding RNA UPK1A-AS1 indicates poor prognosis of hepatocellular carcinoma and promotes cell proliferation through interaction with EZH2[J].J Exp Clin Cancer Res,2020,39(1):229.
10 Gao R,Fang C,Xu J,et al.LncRNA CACS15 contributes to oxaliplatin resistance in colorectal cancer by positively regulating ABCC1 through sponging miR-145[J].Arch Biochem Biophys,2019,663:183-191.
11 He T,Zhang L,Kong Y,et al.Long non-coding RNA CASC15 is upregulated in hepatocellular carcinoma and facilitates hepatocarcinogenesis[J].Int J Oncol,2017,51(6):1722-1730.
12 Li Y,Zhou B,Dai J,et al.Aberrant upregulation of LRRC1 contributes to human hepatocellular carcinoma[J].Mol Biol Rep,2013,40(7):4543-4551.
13 Torre LA,Bray F,Siegel RL,et al.Global cancer statistics,2012[J].CA Cancer J Clin,2015,65(2):87-108.
14 Sheng L,Wei R.Long non-coding RNA-CASC15 promotes cell proliferation,migration,and invasion by activating Wnt/beta-catenin signaling pathway in Melanoma[J].Pathobiology,2020,87(1):20-29.
15 Zhang C,Wang GX,Fu B,et al.LncRNA CASC15 promotes migration and invasion in prostate cancer via targeting miR-200a-3p[J].Eur Rev Med Pharmacol Sci,2019,23(19):8303-8309.
16 Yu L,Xu Q,Yu W,et al.LncRNA cancer susceptibility candidate 15 accelerates the breast cancer cells progression via miR-153-3p/KLF5 positive feedback loop[J].Biochem Biophys Res Commun,2018,506(4):819-825.
17 Huarte M.The emerging role of lncRNAs in cancer[J].Nat Med,2015,21(11):1253-1261.
18 Sun Y,Liu Z,Huang L,et al.MiR-144-3p inhibits the proliferation,migration and invasion of lung adenocargen cancer cells by targeting COL11A1[J].J Chemother,2021,33(6):409-419.
19 Wang P,Yang Z,Ye T,et al.lncTUG1/miR-144-3p affect the radiosensitivity of esophageal squamous cell carcinoma by competitively regulating c-MET[J].J Exp Clin Cancer Res,2020,39(1):7.
20 Xing B,Shen C,Yang Q,et al.miR-144-3p represses hepatocellular carcinoma progression by affecting cell aerobic glycolysis via FOXK1[J].Int J Exp Pathol,2023,104(3):117-127.
21 Martin JW,Yoshimoto M,Ludkovski O,et al.Analysis of segmental duplications,mouse genome synteny and recurrent cancer-associated amplicons in human chromosome 6p21-p12[J].Cytogenet Genome Res,2010,128(4):199-213.
22 Chochi Y,Kawauchi S,Nakao M,et al.A copy number gain of the 6p arm is linked with advanced hepatocellular carcinoma:an array-based comparative genomic hybridization study[J].J Pathol,2009,217(5):677-684.
23 Jia D,Wei L,Guo W,et al.Genome-wide copy number analyses identified novel cancer genes in hepatocellular carcinoma[J].Hepatology,2011,54(4):1227-1236.
24 Li Y,Chen G,Yan Y,et al.CASC15 promotes epithelial to mesenchymal transition and facilitates malignancy of hepatocellular carcinoma cells by increasing TWIST1 gene expression via miR-33a-5p sponging[J].Eur J Pharmacol,2019,860:172589.
25 Gai SY,Yuan ZH.Long non-coding RNA SOX21-AS1 promotes cell proliferation and invasion through upregulating PAK7 expression by sponging miR-144-3p in glioma cells[J].Neoplasma,2020,67(2):333-343.

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长链非编码RNA CASC15对肝细胞癌增殖、侵袭和迁移的影响及作用机制

The effects and mechanism of long non-coding RNA CASC15 on proliferation,invasion and migration of hepatocellular carcinoma

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