食管癌中miRNA枢纽靶基因的鉴定和验证

doi:10.13418/j.issn.1001-165x.2025.1.09

PDF(4657 KB)

中国临床解剖学杂志 ›› 2025, Vol. 43 ›› Issue (1) : 54-62. DOI: 10.13418/j.issn.1001-165x.2025.1.09

实验研究

食管癌中miRNA枢纽靶基因的鉴定和验证

赵婷婷¹，宋爽¹，石科²，李晓英¹，李攀阳¹，李宁宁³*

作者信息 +

Identification and validation of miRNA hub target genes in esophageal cancer

Zhao Tingting ¹, Song Shuang ¹, Shi Ke², Li Xiaoying¹, Li Panyang ¹, Li Ningning³*

Author information +

文章历史 +

摘要

目的探讨与食管癌发生发展相关的miRNAs-mRNAs，研究PALB2对人食管癌EC109细胞生物学性状的影响。方法从GEO和TCGA数据库下载食管癌miRNAs数据集筛选交集，通过starbase数据库预测交集miRNAs的潜在靶基因，并使用PPI网络分析确定枢纽靶基因；对靶基因和枢纽靶基因分别进行GO和KEGG功能富集分析。选择预后差异显著的PALB2基因进行细胞试验，合成PALB2的特异性RNA(si-PALB2)转染EC109细胞，RT-qPCR检测细胞中PALB2的表达量，MTT、Transwell和流式分析PALB2对细胞活力、侵袭迁移和凋亡能力的影响；Western blot检测细胞周期蛋白CyclinD1、凋亡抑制蛋白Bcl-2和基质金属蛋白酶MMP-9的表达变化。结果生物信息学分析预测了参与食管癌发生发展的重要miRNAs-mRNAs，并通过实验验证敲降PALB2的表达后，EC109细胞活力、侵袭转移能力均明显降低，凋亡能力明显增加，cyclinD1、Bcl-2及MMP-9表达均明显下调。结论 MiRNAs-mRNAs是预测食管癌发生发展的生物标志物；PALB2在食管癌中发挥促癌作用，与调控细胞周期蛋白、抗凋亡蛋白及侵袭转移相关蛋白密切相关，是诊断和治疗的潜在靶点。

Abstract

Objective To investigate miRNAs-mRNAs related to the occurrence and development of esophageal cancer, and study the effect of PALB2 on the biological characters of human esophageal cancer EC109 cells. Methods The data of esophageal cancer miRNAs were downloaded from GEO and TCGA databases to screen out the intersecting miRNAs. The potential target genes of intersection were predicted by starbase database, and the key target genes were identified by PPI network analysis. Functional enrichment analysis was performed for differential genes and hub target genes in GO and KEGG databases. PALB2 gene with significant prognostic difference was selected for cell test, and PALB2 specific small interfering RNA (si-PALB2) was synthesized and transfected into EC109 cells. The expression level of PALB2 in the cells was detected by RT-qPCR. The effects of PALB2 on cell viability, invasion, migration and apoptosis were investigated by MTT, Transwell and flow cytometry. The expression of cyclin CyclinD1, apoptosis suppressor Bcl-2 and matrix metalloproteinase MMP-9 were detected by Western blot. Results Bioinformatics analysis showed that miRNAs-mRNAs were involved in the development of esophageal cancer. After PALB2 knockdown, EC109 cell viability, invasion and metastasis ability were significantly decreased, apoptosis ability was significantly increased, and the expressions of cyclinD1, Bcl-2 and MMP-9 were significantly down-regulated. Conclusions MiRNAs-mRNAs are biomarkers for predicting the development of esophageal cancer. PALB2 plays a role in promoting cancer in esophageal cancer, and is closely related to the regulation of cyclin, anti-apoptotic protein and invasion and metastasis related protein, which is a potential target for diagnosis and treatment.

导出引用

赵婷婷, 宋爽, 石科, 李晓英, 李攀阳, 李宁宁. 食管癌中miRNA枢纽靶基因的鉴定和验证[J]. 中国临床解剖学杂志. 2025, 43(1): 54-62 https://doi.org/10.13418/j.issn.1001-165x.2025.1.09

Zhao Tingting, Song Shuang, Shi Ke, Li Xiaoying, Li Panyang, Li Ningning. Identification and validation of miRNA hub target genes in esophageal cancer[J]. Chinese Journal of Clinical Anatomy. 2025, 43(1): 54-62 https://doi.org/10.13418/j.issn.1001-165x.2025.1.09

中图分类号： R735.

参考文献

[1] 姚一菲, 孙可欣, 郑荣寿. 《2022全球癌症统计报告》解读：中国与全球对比[J]. 中国普外基础与临床杂志, 2024, 31(7):769-780. DOI:10.7507/1007-9424.
[2] Bray F, Laversanne M, Sung H, et al. Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J] .CA Cancer J Clin, 2024, 74(3): 229-263. DOI:10.3322/caac.21834.
[3] Diener C, Keller A, Meese E. Emerging concepts of miRNA therapeutics: from cells to clinic[J] .Trends Genet, 2022, 38(6): 613-626. DOI: 10.1016/j.tig.2022.02.006.
[4] 吕艳利, 巴凯, 方政. 微小RNA-3651过表达通过介导核因子κB信号通路抑制人舌癌细胞CAL27的生长与侵袭[J]. 中国临床解剖学杂志, 2023, 41(2):194-199. DOI:10.13418/j.issn.1001-165x.
[5] 陆霞娟, 蒋海峰, 马晶晶. 胃癌黏膜组织miR-127、PALB2表达与临床特征及预后的相关性研究[J].检验医学与临床, 2021,18(12):1738-1742. DOI: 10.3969/j.issn.1672-9455.
[6] 涂媛, 章培, 陈琼, 等. FANCD2、PALB2表达水平与非小细胞肺癌临床特征及预后的关系[J].中国现代医学杂志, 2023,33(18):88-95. DOI: 10.3969/j.issn.1005-8982.
[7] Anbil S, Reiss KA. Targeting BRCA and PALB2 in Pancreatic Cancer[J]. Curr Treat Options Oncol, 2024, 25(3): 346-363. DOI: 10.1007/s11864-023-01174-0.
[8] Du JT, Zhang SL, Zhang XD, et al. miR-1301-3p promotes invasion and migration and EMT progression in esophageal cancer by downregulating NBL1 expression[J]. Thorac Cancer, 2023, 14(30): 3032-3041. DOI: 10.1111/1759-7714.15093.
[9] He B, Zhao Z, Cai Q, et al. MiRNA-based biomarkers, therapies, and resistance in Cancer[J]. International Journal of Biological Sciences, 2020, 16(14):2628-2647. DOI: 10.7150/ijbs.47203.
[10]Foo TK, Xia B. BRCA1-dependent and independent recruitment of PALB2-BRCA2-RAD51 in the DNA damage response and cancer[J]. Cancer Res, 2022, 82(18):3191-3197. DOI: 10.1158/0008-5472.CAN-22-1535
[11]王月, 周亮, 刘丽芳, 等. PALB2是一种与乳腺癌免疫浸润相关的预后生物标志物[J].标记免疫分析与临床, 2023, 30(7):1187-1199. DOI: 10.11748/bjmy.issn.1006-1703.
[12]Zhu YY, Wu F, Hu JN, et al. LDHA deficiency inhibits trophoblast proliferation via the PI3K/AKT/FOXO1/CyclinD1 signaling pathway in unexplained recurrent spontaneous abortion.[J]. FASEB J, 2023. 37(2): e22744. DOI: 10.1096/fj.202201219RR.
[13]Dyer M, Walter HS. BCL2 inhibition: back to the future[J]. Blood, 2024. 143(18): 1787-1788. DOI: 10.1182/blood.2023023796.
[14]Augoff K, Hryniewicz-Jankowska A, Tabola R, et al. MMP9: A tough target for targeted therapy for cancer[J]. Cancers (Basel), 2022, 14(7):1847. DOI: 10.3390/cancers14071847.