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目的 探讨长链非编码RNA(lncRNA)沉默信息调节因子1-动脉粥样硬化(SIRT1-AS)对人结直肠癌(CRC)细胞(HCT116)生物行为的影响及作用机制。方法 成功构建SIRT1-AS过表达HCT116细胞模型后，开展细胞计数试剂盒-8(CCK-8)实验、细胞侵袭(Transwell)实验以及划痕实验，分析SIRT1-AS对HCT116细胞的增殖、侵袭和迁移能力的影响。使用KOBAS 2.0服务器对基因本体(GO)和京都基因与基因组百科全书(KEGG)进行差异表达基因(DEGs)富集分析并通过TNMplot数据库以及人类蛋白质图谱(HPA)分析关键靶基因在肿瘤组织中的表达水平。结果 SIRT1-AS的过表达显著抑制了人CRCHCT116细胞的增殖、侵袭和迁移(P均<0.05)。RNA测序发现，在SIRT1-AS过表达后SIRT1、DHRS2和BHLHE41基因表达明显升高(P均<0.05),而核糖核酸线粒体RNA加工(RMRP)与碳酸酐酶9(CA9)基因表达明显降低(P均<0.05)。SIRT1-AS过表达影响可变剪接事件，涉及调控细胞周期、DNA修复、转录调控、信号转导等关键生物学过程，其中FBXL19、CLK2、NUP62CL、SLAIN2和FMR1等基因的可变剪接事件发生了显著的变化(P均<0.05)。结论 SIRT1-AS在人CRC中发挥抑癌作用，其可能是通过转录调控及影响可变剪接抑制肿瘤进展。
Abstract：Objective To investigate the effect and mechanism of Long Non-coding RNA(lncRNA) SIRT1-AS(Sirtuin 1 Antisense RNA) on the biological behaviors of human Colorectal Cancer(CRC) HCT116 cells. Methods After successfully constructing the SIRT1-AS overexpression HCT116 cell model, Cell Counting Kit-8(CCK-8) assay, Transwell invasion assay and wound healing assay were performed to analyze the effects of SIRT1-AS on the proliferation, invasion and migration abilities of HCT116 cells. Differential expression gene(DEG) enrichment analysis was conducted using the Gene Ontology(GO) terms and Kyoto Encyclopedia of Genes and Genomes(KEGG) through the KOBAS 2.0 server. The expression levels of key target genes in tumor tissues were analyzed using using the TNM plot database and Human Protein Atlas(HPA). Results Overexpression of SIRT1-AS significantly inhibited the proliferation, invasion and migration of human CRC HCT116 cells(all P<0.05). RNA sequencing results showed thatafter SIRT1-AS overexpression, the expression levels of SIRT1, DHRS2 and BHLHE41 genes were significantly increased(all P<0.05), while the expression levels of RMRP(RNA Component of Mitochondrial RNA Processing Endoribonuclease) and CA9(Carbonic Anhydrase 9) genes weresignificantly decreased(all P<0.05). SIRT1-AS overexpression affected alternative splicing events, involving the regulation of key biological processes such as cell cycle regulation, DNA repair, transcriptional regulation and signal transduction. Notably, alternative splicing events of genes including FBXL19, CLK2, NUP62CL, SLAIN2 and FMR1changed significantly(all P<0.05). Conclusion SIRT1-ASexerts a tumor-suppressive effect in human CRC, which may inhibit tumor progression through transcriptional regulation and modulation of alternative splicing.
[1] SAWICKI T,RUSZKOWSKA M,DANIELEWICZ A,et al.A review of colorectal cancer in terms of epidemiology,risk factors,development,symptoms and diagnosis[J].Cancers,2021,13(9):2025.
[2] SIEGEL R L,WAGLE N S,CERCEK A,et al.Colorectal cancer statistics,2023[J].CA,2023,73(3):233-254.
[3] LIN Y,ZHAO W,LV Z,et al.The functions and mechanisms of long non-coding RNA in colorectal cancer[J].Front Oncol,2024,14:1419972.
[4] 向恬，马瑞红，王宝娟，等.SIRT1调控卵泡发育、延缓卵巢衰老的研究进展[J].中国生育健康杂志，2020,31(4):389-391.
[5] HASHEMI M,ZALI N,GHAFARZADEH-DASTJERDI S Z,et al.SIRT1 as a masterful reciprocal regulator of molecular mechanisms and signaling pathways involved in tumor growth and expansion[J].Clin Exp Med,2025,25(1):225.
[6] KIM D,LANGMEAD B,SALZBERG S L.HISAT:a fast spliced aligner with low memory requirements[J].Nat Methods,2015,12(4):357-360.
[7] TRAPNELL C,WILLIAMS B A,PERTEA G,et al.Transcript assembly and quantification by RNA-Seq reveals unannotated transcripts and isoform switching during cell differentiation[J].Nat Biotechnol,2010,28(5):511-515.
[8] LOVE M I,HUBER W,ANDERS S.Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2[J].Genome Biol,2014,15(12):550.
[9] JIN L,LI G,YU D,et al.Transcriptome analysis reveals the complexity of alternative splicing regulation in the fungus verticillium dahliae[J].BMC Genomics,2017,18(1):130.
[10] XIE C,MAO X,HUANG J,et al.KOBAS 2.0:a web server for annotation and identification of enriched pathways and diseases[J].Nucleic Acids Res,2011,39:316-322.
[11] 张蕾，胡尔西旦·尼牙孜，赵化荣，等.长链非编码MALAT1在宫颈癌转移中的作用[J].新疆医科大学学报，2018,41(3):338-341,344.
[12] WANG Y,PANG W,WEI N,et al.Identification,stability and expression of sirt1 antisense long non-coding RNA[J].Gene,2014,539(1):117-124.
[13] HUSSEIN N A,EBIED S A,BELAL A A M,et al.Expression profiling of circulating lncRNA GIAT4RA,lncRNA AATBC,lncRNA SIRT1-AS,and SMARCB1 in lung cancer patients[J].Bmc Cancer,2024,24(1):1175.
[14] LI B,HU Y,LI X,et al.Sirt1 antisense long noncoding RNA promotes cardiomyocyte proliferation by enhancing the stability of sirt1[J].J AM Heart Assoc,2018,7(21):e9700.
[15] WEI Z,XIA J,LI J,et al.SIRT1 promotes glucolipid metabolic conversion to facilitate tumor development in colorectal carcinoma[J].Int J Biol Sci,2023,19(6):1925-1940.
[16] WANG X,JIANG Y,YE W,et al.SIRT1 promotes the progression and chemoresistance of colorectal cancer through the p53/miR-101/KPNA3 axis[J].Cancer Biol Ther,2023,24(1):2235770.
[17] YUAN J,MINTER-DYKHOUSE K,LOU Z.A c-Myc-SIRT1 feedback loop regulates cell growth and transformation[J].J Cell Biol ,2009,185(2):203-211.
[18] BRANDL L,KIRSTEIN N,NEUMANN J,et al.The c-MYC/NAMPT/SIRT1 feedback loop is activated in early classical and serrated route colorectal cancer and represents a therapeutic target[J].Med Oncol,2018,36(1):5.
[19] SHAFQAT N,SHAFQAT J,EISSNER G,et al.Hep27,a member of the short-chain dehydrogenase/reductase family,is an NADPH-dependent dicarbonyl reductase expressed in vascular endothelial tissue[J].CMLS,2006,63(10):1205-1213.
[20] LUO X,LI N,ZHAO X,et al.DHRS2 mediates cell growth inhibition induced by Trichothecin in nasopharyngeal carcinoma[J].CR,2019,38(1):300.
[21] HAO Q,ZHOU X.The emerging role of long noncoding RNA RMRP in cancer development and targeted therapy[J].Cancer Biol Med,2022,19(2):140-146.
[22] WANG C,ZHAO N,ZHENG Q,et al.BHLHE41 promotes U87 and U251 cell proliferation via ERK/cyclinD1 signaling pathway[J].Cancer Manag Res,2019,11:7657-7672.
[23] CHEN S,DONG Q,WAN Z,et al.BHLHE41 overexpression alleviates the malignant behavior of colon cancer cells induced by hypoxia via modulating HIF-1α/EMT pathway[J].Gastroent Res Pract,2022,2022:6972331.
[24] 许建昆，周露婷，张文净，等.CA9在透明细胞肾细胞癌预后评估中的价值[J].诊断学理论与实践，2023,22(1):37-43.
[25] WANG G,CHENG Z,LIU F,et al.CREB is a key negative regulator of carbonic anhydrase IX (CA9) in gastric cancer[J].Cell Signal,2015,27(7):1369-1379.
[26] LIN C,LAI S,SHEN C,et al.Fenofibrate inhibits hypoxia-inducible factor-1 alpha and carbonic anhydrase expression through activation of AMP-activated protein kinase/HO-1/Sirt1 pathway in glioblastoma cells[J].Environ Toxicol,2021,36(12):2551-2561.
[27] DONG W,LU J,LI Y,et al.SIRT1:a novel regulator in colorectal cancer[J].Biomed Pharmacother,2024,178:117176.
[28] CHENG L,WU G,YAO W,et al.Nuclear-localized SIRT1 inhibits apoptosis via deacetylating p53[J].Int J Biochem Cell Biol,2025,187:106841.
[29] ZOU L,ZHANG M,HE S,et al.Genetic variants of lncRNA associated with splicing regulation and their impact on ovarian cancer development[J].Fun Int Genomic,2025,25(1):185.
[30] LIU J,WU W,JIN J.A novel mutation in SIRT1-AS leading to a decreased risk of HCC[J].Oncol Rep,2015,34(5):2343-2350.
基本信息:
中图分类号:R735.34
引用信息:
[1]刘洋,郑高翔,刘林.lncRNA SIRT1-AS在人结直肠癌细胞中的作用机制研究[J].新疆医科大学学报,2026,49(03):331-338.
基金信息:
新疆维吾尔自治区自然科学基金青年科学基金项目(2022D01C799)
2026-03-15
2026-03-15