实用肿瘤杂志2025，Vol.40Issue(3)：218-226,9.DOI:10.13267/j.cnki.syzlzz.2025.034
神经肽FF通过调控RhoA/ROCK/YAP信号通路促进结直肠癌转移的分子机制研究
Molecular mechanism of neuropeptide FF promoting metastasis in colorectal cancer by regulating RhoA/ROCK/YAP signaling pathway
摘要
Abstract
Objective To explore the molecular mechanism by which neuropeptide FF(NPFF)promotes colorectal cancer metastasis through activating the Ras homolog family member A(RhoA)/Rho associated coiled-coil containing protein kinase(ROCK)/Yes-associated protein(YAP)signaling pathway.Methods The expression of NPFF in colorectal cancer and its prognostic value were analyzed via The Cancer Genome Atlas(TCGA)public database.The mRNA expression of NPFF in colorectal cancer cells and intestinal epithelium normal cell NCM460 was detected by real-time quantitative polymerase chain reaction(RT-qPCR).Small interfering RNA and plasmid trans-fection were used to construct cell lines with upregulated and downregulated NPFF.The protein levels of RhoA,YAP,and phospho-YAP were detected by Western blotting.The migration ability of colorectal cancer cells with NPFF knockdown/overexpression was detected by transwell assay.The subcellular localization of YAP was observed by immunofluorescence staining.Results The TCGA database and RT-qPCR results indicated that NPFF was highly expressed in colorectal cancer tissues and colorectal cancer cells including DiFi,DLD-1,SW620,KM12C,HT29,SW480,HCT8,and KM12SM cells(all P<0.05),and was associated with poor prognosis.The transwell assay showed that the migration ability of SW480 and KM12C cells was significantly weakened after NPFF knockdown(both P<0.01),while the migration of DiFi and DLD-1 cells was significantly enhanced after NPFF overexpression(both P<0.01).The treatment of 5 μmol/L Y27632,a ROCK 1/2 inhibitor,for 24 h effectively counteracted the enhanced migration of NPFF-overexpressed DLD-1 and DiFi cells(both P<0.01).RhoA activity detection indicated that the level of RhoA activation in SW480 and DiFi cells significantly increased after the addition of 3 μmol/L NPFF recombinant peptide for 30 min(both P<0.05).Western blotting showed that YAP in SW480 and DiFi cells was significantly dephosphorylated after the treatment of 3 μmol/L NPFF recombinant peptide for 3,6,and 12 h(all P<0.05).Immunofluores-cence staining showed that the fluorescence intensity of actin and the nuclear localization of YAP increased after the treatment of 3 μmol/L NPFF recombinant peptide for 3 h in DiFi cells.Nuclear-cytoplasmic protein separation detection found that YAP significantly translocat-ed from cytoplasm to nucleus after the treatment of 3 μmol/L NPFF recombinant peptide for 3 and 6 h in SW480 and DiFi cells(all P<0.05).RT-qPCR indicated that NPFF upregulated the expression of the downstream target genes of YAP including connective tissue growth factor(CTGF),cysteine rich angiogenic inducer 61(CYR61),angiomotin like 2(AMOTL2),and ankyrin repeat domain 1(ANKRD1)in SW480 and DiFi cells(all P<0.05).Conclusions NPFF promotes the metastasis in colorectal cancer by activating the RhoA/ROCK/YAP signaling pathway.关键词
结直肠癌/肿瘤转移/神经肽FF/Ras同源家族成员A/Rho相关卷曲螺旋蛋白激酶/Yes相关蛋白Key words
colorectal cancer/tumor metastasis/neuropeptide FF/Ras homolog family member A/Rho associated coiled-coil containing protein kinase/Yes-associated protein引用本文复制引用
王好甲,苗格,赵晓迪,王新..神经肽FF通过调控RhoA/ROCK/YAP信号通路促进结直肠癌转移的分子机制研究[J].实用肿瘤杂志,2025,40(3):218-226,9.基金项目
国家自然科学基金面上项目(82173256) （82173256）
陕西省重点研发计划(2024SF-GJHX-04) （2024SF-GJHX-04）
消化系肿瘤整合防治全国重点实验室课题(CBSKL2022ZZ55) （CBSKL2022ZZ55）
