| 319 | 0 | 115 |
| 下载次数 | 被引频次 | 阅读次数 |
目的 探讨锌指蛋白家族成员锌指蛋白473(ZNF473)对肝细胞癌（LIHC）增殖、迁移的影响及潜在作用机制。方法 通过癌症基因组图谱（TCGA）数据库分析ZNF473在泛癌、LIHC组织与正常组织中的表达水平；通过Kaplan Meier Plotter数据库Log-rank检验分析ZNF473对LIHC患者预后影响；以HepG2、MHCC97-H细胞为研究对象，分阴性对照组（NC）和实验组（siZNF473），在HepG2、MHCC97-H中转染ZNF473干扰载体，采用CCK-8、Transwell实验及划痕愈合实验检测ZNF473对HepG2、MHCC97-H细胞增殖和迁移的影响；采用Western blot检测敲低ZNF473后LIHC细胞中磷酸化蛋白激酶B(p-AKT)、c-Myc蛋白、蛋白激酶B(AKT)等蛋白表达的变化。细胞实验所有数据均使用GraphPad Prism 9.0软件进行分析。结果 TCGA数据库分析结果显示，ZNF473在包括LIHC在内的17种癌症中高表达。人蛋白图谱（HPA）数据库中的免疫组织化学（IHC）染色提示，ZNF473在LIHC组织中高表达。单细胞测序数据分析显示，ZNF473在恶性细胞亚群中的表达量高于其他细胞亚群。Kaplan-Meier Plotter数据库对LIHC患者的生存分析显示，ZNF473高表达组的总生存期（OS）、疾病特异性生存期（DSS）、无进展生存期（PFS）及无复发生存期（RFS）均显著短于低表达组。CCK-8实验表明敲低ZNF473后，HepG2、MHCC97-H细胞增殖能力下降；划痕愈合实验和Transwell实验结果显示，敲低ZNF473后HepG2、MHCC97-H细胞迁移能力下降。RT-qPCR结果显示，si-ZNF473组磷酸酶与张力蛋白同源物（PTEN）相对表达量差异无统计学意义，磷脂酰肌醇-4,5-二磷酸3-激酶催化亚基α(PIK3CA)、磷酸肌醇-3-激酶调节亚基1(PIK3R1)表达差异有统计学意义（P<0.05）。Western blot结果显示，下调ZNF473表达后p-AKT、c-Myc蛋白表达水平下降（P<0.05）。结论 ZNF473在LIHC细胞和组织中呈高表达，ZNF473表达可能通过调控PI3K/AKT/c-Myc通路促进LIHC细胞增殖、迁移。
Abstract：Objective To investigate the effects of zinc finger protein 473(ZNF473), a member of the zinc finger protein family, on the proliferation and migration of liver hepatocellular carcinoma(LIHC) cells and to explore its potential underlying mechanism.Methods The expression levels of ZNF473 in pan-cancer and in HCC tissues versus normal tissues were analyzed using The Cancer Genome Atlas(TCGA) database. The prognostic significance of ZNF473 in patients with HCC was evaluated using the Kaplan-Meier Plotter database with the Log-rank test. Human HCC cell lines HepG2 and MHCC97-H were used as experimental models and divided into a negative control(NC) group and an experimental group(si-ZNF473). ZNF473 interference vectors were transfected into HepG2 and MHCC97-H cells. CCK-8 assays, Transwell assays, and wound-healing assays were performed to assess the effects of ZNF473 on the proliferation and migration of HepG2 and MHCC97-H cells. Western blot was used to detect changes in the expression levels of phosphorylated protein kinase B(p-AKT), c-Myc, AKT, and other related proteins in HCC cells after ZNF473 knockdown. All cell experiment data were analyzed using GraphPad Prism 9. 0 software.Results TCGA database analysis showed that ZNF473 was highly expressed in 17 cancer types, including LIHC. Immunohistochemical(IHC) staining data from the Human Protein Atlas(HPA) database indicated that ZNF473 was highly expressed in LIHC tissues. Single-cell RNA sequencing analysis showed that ZNF473 expression was higher in the malignant cell subpopulation than in other cell subpopulations. Survival analysis of patients with LIHC using the Kaplan-Meier Plotter database demonstrated that the high-ZNF473-expression group had significantly shorter overall survival(OS), disease-specific survival(DSS), progression-free survival(PFS), and recurrence-free survival(RFS) than the low-expression group. CCK-8 assays showed that knockdown of ZNF473 reduced the proliferative capacity of HepG2 and MHCC97-H cells. Wound-healing and Transwell assays demonstrated that ZNF473 knockdown suppressed the migratory ability of HepG2 and MHCC97-H cells. RT-qPCR results showed no statistically significant difference in phosphatase and tensin homolog(PTEN) relative expression in the si-ZNF473 group, whereas the expression levels of PIK3 CA and PIK3R1 were significantly altered(P<0. 05). Western blot analysis showed that downregulation of ZNF473 significantly decreased the protein expression levels of p-AKT and c-Myc(P<0. 05).Conclusion ZNF473 is highly expressed in LIHC cells and tissues. ZNF473 may promote the proliferation and migration of LIHC cells by regulating the PI3K/AKT/c-Myc signaling pathway.
[1]Wang J L, Wang J, Tian C Y. Evolution of KRAB-containing zinc finger proteins and their roles in species evolution[J]. Hereditas, 2016, 38(11):971-8. doi:10. 16288/j. yczz. 16-056.
[2]Bulanenkova S S, Filyukova O B, Snezhkov E V, et al. Suppression of the testis-specific transcription of the ZBTB32 and ZNF473 genes in germ cell tumors[J]. Acta Naturae, 2022, 14(3):85-94. doi:10. 32607/actanaturae. 11620.
[3]Glaviano A, Foo A S C, Lam H Y, et al. PI3K/AKT/mTOR signaling transduction pathway and targeted therapies in cancer[J].Mol Cancer, 2023, 22(1):138. doi:10. 1186/s12943-023-01827-6.
[4]王菊琴，何淋玲，沈钰娜，等. PLVAP通过PI3K/AKT通路促进肝细胞癌增殖、迁移和侵袭[J].安徽医科大学学报，2025,60(5):860-8. doi:10. 19405/j. cnki. issn1000-1492. 2025. 05. 012.Wang J Q, He L L, Shen Y N, et al. PLVAP promotes proliferation, migration and invasion of hepatocellular carcinoma via PI3K/AKT pathway[J]. Acta Univ Med Anhui, 2025, 60(5):860-8.doi:10. 19405/j. cnki. issn1000-1492. 2025. 05. 012.
[5]胡磊，陈红霞，周钢桥. METTL16对肝癌细胞增殖、迁移和侵袭能力的影响及其作用机制[J].安徽医科大学学报，2022, 57(12):1849-57. doi:10. 19405/j. cnki. issn1000-1492. 2022. 12. 001.Hu L, Chen H X, Zhou G Q. Effects of METTL16 on proliferation,migration and invasion of hepatocellular carcinoma cells and its mechanism of action[J]. Acta Univ Med Anhui, 2022, 57(12):1849-57. doi:10. 19405/j. cnki. issn1000-1492. 2022. 12. 001.
[6]Wagner E J, Marzluff W F. ZFP100, a component of the active U7 snRNP limiting for histone pre-mRNA processing, is required for entry into S phase[J]. Mol Cell Biol, 2006, 26(17):6702-12. doi:10. 1128/MCB. 00391-06.
[7]Ai X, Xiang L, Huang Z, et al. Overexpression of PIK3R1 promotes hepatocellular carcinoma progression[J]. Biol Res, 2018,51(1):52. doi:10. 1186/s40659-018-0202-7.
[8]He S J, Zhang J Y, Zhang W, et al. FOXA1 inhibits hepatocellular carcinoma progression by suppressing PIK3R1 expression in male patients[J]. J Exp Clin Cancer Res, 2017, 36(1):175.doi:10. 1186/s13046-017-0646-6.
[9]Sadrkhanloo M, Paskeh M D A, Hashemi M, et al. New emerging targets in osteosarcoma therapy:PTEN and PI3K/Akt crosstalk in carcinogenesis[J]. Pathol Res Pract, 2023, 251:154902. doi:10. 1016/j. prp. 2023. 154902.
[10]Mukherjee R, Vanaja K G, Boyer J A, et al. Regulation of PTEN translation by PI3K signaling maintains pathway homeostasis[J].Mol Cell, 2021, 81(4):708-23. e5. doi:10. 1016/j. molcel.2021. 01. 033.
[11]Gregory M A, Qi Y, Hann S R. Phosphorylation by glycogen synthase kinase-3 controls c-myc proteolysis and subnuclear localization[J]. J Biol Chem, 2003, 278(51):51606-12. doi:10. 1074/jbc. M310722200.
[12]Wei W, Zhao X, Wu S, et al. Dihydroartemisinin triggers c-Myc proteolysis and inhibits protein kinase B/glycogen synthase kinase3β pathway in T-cell lymphoma cells[J]. Oncol Lett, 2018, 16(5):6838-46. doi:10. 3892/ol. 2018. 9450.
[13]Zhao M, Zhang X L, Chen H Z, et al. The clinicopathologic and prognostic significance of c-myc expression in hepatocellular carcinoma:a meta-analysis[J]. Front Bioinform, 2021, 1:706835. doi:10. 3389/fbinf. 2021. 706835.
[14]Lin C P, Liu C R, Lee C N, et al. Targeting c-Myc as a novel approach for hepatocellular carcinoma[J]. World J Hepatol, 2010,2(1):16-20. doi:10. 4254/wjh. v2. i1. 16.
[15]Wang H, Xu H, Ma F, et al. Zinc finger protein 703 induces EMT and sorafenib resistance in hepatocellular carcinoma by transactivating CLDN4 expression[J]. Cell Death Dis, 2020, 11(4):225. doi:10. 1038/s41419-020-2422-3.
基本信息:
DOI：10.19405/j.cnki.issn1000-1492.2026.08.009
中图分类号:R735.7
引用信息:
[1]刘闯,任益凡,王妍,等.ZNF473通过PI3K/AKT/c-Myc通路促进肝细胞癌的增殖与迁移[J].安徽医科大学学报,2026,61(08):1400-1407.DOI:10.19405/j.cnki.issn1000-1492.2026.08.009.
基金信息:
国家自然科学基金项目（编号：82470693）; 山西省重点实验室项目（编号：202204010931008）; 山西省重点研发计划项目（编号：202302130501013）~~
2026-07-13
2026-07-13
2026-07-13