摘要
目的探讨miR-506对恶性黑色素瘤(MM)的生物学行为、间叶上皮转化(MET)及DNA损伤同源重组通路的影响。方法采用qRT-PCR检测48例新鲜MM组织中miR-506的表达水平。应用IHC检测147例MM患者组织中MET及DNA损伤相关蛋白的表达水平。选取人MM细胞系A375和A875进行细胞功能实验。过表达或抑制miR-506后,通过MTT、Transwell小室等细胞功能实验来验证miR-506对MM细胞的增殖、迁移及侵袭性的影响,并使用免疫印迹分析来进一步观察miR-506对MET相关蛋白及DNA损伤同源重组相关蛋白表达的影响。采用Kaplan-Meier法进行生存分析。结果miR-506在转移灶中低表达,在原发灶中高表达,其高表达与患者更长的总生存(OS)(P=0.002)、无病生存(DFS)(P<0.001)和无进展生存(PFS)(P=0.041)相关。间叶组织标志蛋白N-cadherin高表达预示较差的OS(P=0.026),上皮组织标志蛋白E-cadherin高表达预示较好的OS(P=0.006)。DNA损伤同源重组相关蛋白RAD51和ATM高表达均与较差的OS相关(RAD51:P=0.014;ATM:P=0.002),而RAD52高表达与较好的OS相关(P=0.006)。Spearman相关分析结果显示miR-506与Vimentin(r=-0.38,P=0.008)、Slug(r=-0.36,P=0.012)的表达呈负相关,与RAD51呈负相关(r=-0.47,P=0.001)。细胞功能学结果显示过表达miR-506可抑制恶性黑色素瘤细胞的增殖、迁移和侵袭。过表达miR-506可降低恶性黑色素瘤细胞的N-cadherin、Vimentin、Slug和RAD51的表达。结论miR-506、MET相关蛋白和DNA损伤同源重组相关蛋白均与恶性黑色素瘤患者的预后相关。miR-506作为一个抑癌基因,可能通过调节MET相关蛋白和DNA损伤同源重组通路来抑制黑色素瘤细胞的增殖、迁移和侵袭。
Objective To investigate the biological function of miR-506 in malignant melanoma and its regulatory mechanism on mesenchymal epithelial transition(MET)and DNA damage homologous recombination repair pathway.Methods The expression of miR-506 in 48 cases of fresh malignant melanoma tissues was detected by qRT-PCR.The expression of MET and DNA damage-related proteins in the tissues of 147 patients with malignant melanoma were detected by IHC.The human malignant melanoma cell lines A375 and A875 were used for assessing cell function.Following overexpression or inhibition of miR-506,cell function assays such as MTT and Transwell chamber experiments were conducted to verify its effects on the proliferation,migration,and invasion of malignant melanoma cells.Western blot analysis was used to further examine the impact of miR-506 on the expression of MET-related proteins and homologous recombination pathways of DNA damage.Survival analysis using Kaplan Meier method.Results The expression levels of miR-506 were low in metastatic lesions and high in primary lesions.Its high expression was associated with longer overall survival(OS)(P=0.002),disease-free survival(DFS)(P<0.001)and progression-free survival(PFS)(P=0.041).High expression levels of mesenchymal marker N-cadherin indicated poorer overall survival(P=0.026),while high expression levels of epithelial marker E-cadherin indicated better overall survival(P=0.006).High expression levels of DNA damage homologous recombination related proteins RAD51 and ATM were associated with poor OS(RAD51:P=0.014,ATM:P=0.002),while high expression of RAD52 was associated with good OS(P=0.006).Spearman correlation analysis showed that miR-506 was negatively correlated with the expression of Vimentin and Slug(r=-0.38,P=0.008;r=-0.36,P=0.012),and also negatively correlated with RAD51(r=-0.47,P=0.001).Functional cell assays showed that overexpression of miR-506 inhibited the proliferation,migration,and invasion of malignant melanoma cells.Overexpression of miR-506 decreased the expression of N-cadherin,Vimentin,Slug,and RAD51.Conclusions MiR-506,MET-associated proteins,and DNA damage homologous recombination-associated proteins are associated with the prognosis of patients with malignant melanoma.As a tumor suppressor gene,miR-506 may inhibit the proliferation,migration,and invasion of melanoma cells by regulating MET-related proteins and the homologous recombination repair pathway.
作者
李婷
向俐洁
付来华
郝梦泽
刘新月
杨吉龙
LI Ting;XIANG Lijie;FU Laihua;HAO Mengze;LIU Xinyue;YANG Jilong(Departments of Bone and Soft Tissue Tumor,Tianjin Medical University Cancer Institute and Hospital,Tianjin 300060,China;National Clinical Research Center for Cancer,Tianjin′s Clinical Research Center for Cancer,Key Laboratory of Prevention and Therapy,State Key Laboratory of Druggability Evaluation and Systematic Translational Medicine,Key Laboratory of Molecular Cancer Epidemiology,Tianjin 300060,China;National Cancer Center,National Clinical Research Center for Cancer,Cancer Hospital&Shenzhen Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Shenzhen 518116,China;Hematopoietic Stem Cell Transplantation Center,Institute of Hematology,Blood Diseases Hospital,Chinese Academy of Medical Sciences&Peking Union Medical College,Tianjin 300000,China)
出处
《皮肤性病诊疗学杂志》
2025年第7期463-475,共13页
Journal of Diagnosis and Therapy on Dermato-venereology
基金
国家自然科学基金面上项目(82473477)。
