| 323 | 1 | 142 |
| 下载次数 | 被引频次 | 阅读次数 |
本研究主要探讨环状RNA(circular RNA, circRNA)在鳖甲煎丸抑制肝癌进展过程中的表达变化及分子机制。利用find＿circ和CIRI算法、 KEGG通路和GSEA富集分析,对circRNA的表达及特征进行深入研究。采用彗星实验检测鳖甲煎丸含药血清对肝癌细胞DNA损伤的影响;通过β-半乳糖苷酶染色评估其对肝癌细胞衰老的诱导作用;采用划痕实验检测细胞迁移能力的变化。此外,还构建了肝癌细胞皮下移植动物模型,并观察了瘤体体积和生长曲线的变化。共鉴定出1 764个差异表达的circRNA,其靶基因主要富集于自噬-动物、脊髓小脑共济失调、内质网中的蛋白质加工和细胞衰老等信号通路。GSEA分析进一步显示,这些circRNA的靶基因在UV响应DN、有丝分裂纺锤体和G2M检查点通路中显著富集。彗星实验结果表明,鳖甲煎丸含药血清可显著诱导肝癌细胞的DNA损伤(P<0.01); β-半乳糖苷酶染色实验显示,经干预后的衰老肝癌细胞数量显著增加(P<0.001)。划痕实验结果表明,经干预后的肝癌细胞迁移能力在48 h时被显著抑制(P<0.000 1)。小鼠皮下瘤模型实验结果显示,经鳖甲煎丸灌胃后的肿瘤体积明显减小,生长曲线显著降低(P<0.000 1)。综合以上结果,鳖甲煎丸通过调控circRNA的表达、诱导肝癌细胞的衰老进而抑制肝癌细胞的迁移和增殖以发挥抗肝癌作用。
Abstract：This study focuses on the expression changes and molecular mechanisms of circular RNA(circRNA) in the inhibition of hepatocellular carcinoma development by Biejia Jianwan. The expression and characteristics of circRNA were analyzed using find＿circ and CIRI algorithm, KEGG pathway, and GSEA enrichment. The comet assay was used to detect the effect of Biejia Jianwan drugcontaining serum on DNA damage in hepatocellular carcinoma cells. At the same time, β-galactosidase staining was assessed to evaluate its role in inducing senescence in hepatocellular carcinoma cells. The scratch assay was also used to detect the changes in the migratory ability of cells. In addition, a subcutaneous tumor model of hepatocellular carcinoma cells was established to observe changes in tumor volume and growth curve. The results indicated the identification of 1 764 differentially expressed circRNA, with the target genes mainly enriched in pathways related to autophagy-animal, spinocerebellar ataxia, protein processing in endoplasmic reticulum, and cellular senescence. GSEA analysis further showed that the target genes of these circRNA were significantly enriched in the UV response DN, mitotic spindle, and G2M checkpoint pathways. The results of comet assay showed that Biejia Jianwan drug-containing serum significantly induced DNA damage in hepatocellular carcinoma cells(P<0. 01); β-galactosidase staining assay showed that the number of senescent hepatocellular carcinoma cells was significantly increased after the intervention(P<0. 001). The results of the scratch experiment showed that the migration ability of intervened hepatocellular carcinoma cells was significantly inhibited at 48 h(P<0. 000 1).The experimental results of the mouse subcutaneous tumor model showed that the tumor volume was significantly reduced and the growth curve was decreased considerably after gavage of turtle shell decoction(P<0. 000 1). Taken together, Biejia Jianwan exerts its antihepatocellular carcinoma effects by regulating the expression of circRNA, inducing senescence in hepatocellular carcinoma cells, and subsequently inhibiting their migration and proliferation capabilities.
董明新,孙晓辉,徐畅,等,2021.DNA损伤修复与细胞周期阻滞.国际生物医学工程杂志,44(4):329-333.[DONGX,SUN X H,XU C,et al.,2021.DNA damage repair and cell cycle arrest.International Journal of Biomedical Engineering,44(4):329-333.]
胡卫敏,郭方,2020.鳖甲煎丸治疗慢性乙型肝炎早期肝硬化效果观察.现代中西医结合杂志,29(35):3944-3948.[HU W M,GUO F,2020.Observation on the effect of Biejiajian Pill on early cirrhosis of chronic hepatitis B.Modern Journal of Integrated Traditional Chinese and Western Medicine,29(35):3944-3948.]
李明芬,潘爱萍,林英辉,等,2021.鳖甲煎丸含药血清抑制肝癌细胞的miRNA-mRNA调控机制分析.中成药,43(6):1437-1444.[LI M F,PAN A P,LIN Y H,et al.,2021.Mechanistic analysis of serum Biejiajianwan-induced miRNA-mRNA inhibition in hepatoma carcinoma cells.Chinese Traditional Patent Medicine,43(6):1437-1444.]
林洪升,韦巍,林英辉,等,2021.鳖甲煎丸通过LncRNA-m RNA顺式调控抑制肝癌细胞.基因组学与应用生物学,40(4):1829-1837.[LIN H S,WEI W,LIN Y H,et al.,2021.Liver cancer cells were inhibited through Cis-regulation of LncRNA-mRNA by Biejiajianwan.Genomics and Applied Biology,40(4):1829-1837.]
苗淑莹,杨军,管文燕,等,2023.环状RNA在肝细胞癌发生发展及诊疗中的作用.临床肝胆病杂志,39(8):1983-1991.[MIAO S Y,YANG J,GUAN W Y,et al.,2023.Role of circular RNA in the development,progression,diagnosis,and treatment of hepatocellular carcinoma.Journal of Clinical Hepatology,39(8):1983-1991.]
彭涛,2020.鳖甲煎丸在肝癌治疗中的临床应用.中西医结合肝病杂志,30(6):481-483.[PENG T,2020.Clinical application of Biejiajian Pill in the treatment of liver cancer.Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases,30(6):481-483.]
孙海涛,文彬,陈冠新,等,2018.鳖甲煎丸对肝纤维化模型大鼠肝组织中Wnt/β-catenin信号通路相关蛋白及其靶基因表达的影响.中医杂志,59(10):876-881.[SUN HT,WEN B,CHEN G X,et al.,2018.Effect of Biejiajian Pill on Wnt/β-catenin pathway associated protein and its target gene expression in liver tissue of hepatic fibrosis model rats.Journal of Traditional Chinese Medicine,59(10):876-881.].
卫健委医政司,2024.原发性肝癌诊疗指南(2024年版).中国临床医学,31(2):277-334.[Department of Medical Administration,National Health Commission of the People′Republic of China,2024.Guidelines for the diagnosis and treatment of primary liver cancer (2024 edition).China Clinical Medicine,31(2):277-334.]
谢杨益,林洪升,孔茵芝,等,2022.鳖甲煎丸含药血清对肝癌干细胞的抑制作用及潜在调控机制研究.吉林中医药,42(5):574-578.[XIE Y Y,LIN H S,KONG Y Z,et al.,2022.Study on the inhibitory effect and its potential mechanism of the drug serum of Bijiajian Pills on liver cancer stem cells.Jilin Journal of Chinese Medicine,42 (5):574-578.]
杨利,林洪升,李屏,等,2023.可变剪切事件在鳖甲煎丸抑制肝癌细胞增殖过程中的作用机制及变化.重庆医学,52(14):2101-2106.[YANG L,LIN H S,LI P,et al.,2023.Action mechanism and change of alternative splicing event in Biejiajian Pills inhibiting hepatocellular carcinoma cell proliferation process.Chongqing Medicine,52 (14):2101-2106.]
BRAY F,LAVERSANNE M,SUNG H,et al.,2024.Global cancer statistics 2022:GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries.CACancer J.Clin.,74(3):229-263.
FEI D,WANG F,WANG Y H,et al.,2024.Circular RNAACVR2A promotes the progression of hepatocellular carcinoma through mir-511-5p targeting PI3K-Akt signaling pathway.Mol.Cancer,23(1):159.
KAYA V O,ADEBALI O,2025.UV-induced reorganization of3D genome mediates DNA damage response.Nat.Commun.,16(1):1376.
LI F M,LIU P Y,MI W,et al.,2024.Blocking methionine catabolism induces senescence and confers vulnerability to GSK3 inhibition in liver cancer.Nat.Cancer,5(1):131-146.
NIE P Q,ZHANG C,WU F Y,et al.,2024.The compromised Fanconi anemia pathway in prelamin A-expressing cells contributes to replication stress-induced genomic instability.Adv.Sci.,11(30):e2307751.
QIU H B,CAO S M,XU R H,2021.Cancer incidence,mortality,and burden in China:a time-trend analysis and comparison with the United States and United Kingdom based on the global epidemiological data released in 2020.Cancer Commun.,41(10):1037-1048.
TANG C W,ZHUANG H K,WANG W T,et al.,2024.CircNUP54 promotes hepatocellular carcinoma progression via facilitating HuR cytoplasmic export and stabilizing BIRC3m RNA.Cell Death Dis.,15(3):191.
基本信息:
DOI：10.13417/j.gab.044.000511
中图分类号:R285.5
引用信息:
[1]林洪升,谭金娜,方巧玲,等.鳖甲煎丸抑制肝癌细胞的环状RNA表达谱分析[J].基因组学与应用生物学,2025,44(05):511-519.DOI:10.13417/j.gab.044.000511.
基金信息:
广西自然科学基金项目(2021GXNSFBA220073)资助
2025-04-09
2025-04-09
2025-04-09