1.重庆大学附属人民医院中心实验室,重庆
2.陆军军医大学(第三军医大学)第一附属医院江北院区(中国人民解放军第 958 医院)皮肤科,重庆
3.重庆医科大学附属第二医院泌尿肾病中心,重庆
4.陆军军医大学(第三军医大学)第二附属医院胸外科,重庆
ZHENG Hong,E-mail:ziecoe@tmmu.edu.cn
Received:03 November 2025,
Revised:2026-01-24,
Published:30 March 2026
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谢罗樱子,何志强,张伟楠,等.基于帕金森病相关基因构建黑色素瘤预后模型:七基因特征及其对预后评估与疗效预测的效能验证[J].陆军军医大学学报,2026,48(06):793-808.DOI:10.16016/j.2097-0927.202511006.
XIE L Y Z,HE Z Q,ZHANG W N,et al.Construction of a melanoma prognostic model based on Parkinson’s disease-related genes: a 7-gene signature and validation of its efficacy for prognostic assessment and treatment response prediction[J].J Army Med Univ,2026,48(06):793-808.DOI:10.16016/j.2097-0927.202511006.
谢罗樱子,何志强,张伟楠,等.基于帕金森病相关基因构建黑色素瘤预后模型:七基因特征及其对预后评估与疗效预测的效能验证[J].陆军军医大学学报,2026,48(06):793-808.DOI:10.16016/j.2097-0927.202511006. DOI:
XIE L Y Z,HE Z Q,ZHANG W N,et al.Construction of a melanoma prognostic model based on Parkinson’s disease-related genes: a 7-gene signature and validation of its efficacy for prognostic assessment and treatment response prediction[J].J Army Med Univ,2026,48(06):793-808.DOI:10.16016/j.2097-0927.202511006. DOI:
目的
2
探索帕金森病家族基因的遗传特征,构建帕金森家族基因特征模型,以指导皮肤黑色素瘤(cutaneous melanoma, CMM)患者的预后和治疗。
方法
2
①通过分析TCGA和GTEx数据库,比较帕金森病相关家族基因在黑色素瘤组织和正常组织中的表达差异,通过GEPIA2数据库分析相关基因在不同TNM分期和Clark分级中的表达差异。②基于TCGA数据库,利用预后森林图、LASSO回归模型和风险因素分析,建立包含7个关键基因的特征模型。对该模型进行内部和外部验证,并对模型基因进行GO富集和KEGG通路富集分析。③基于TIMER数据库,分析预后模型中基因突变对肿瘤微环境(tumor microenvironment, TME)免疫细胞浸润的影响,利用ssGSEA热图分析高风险组与低风险组间免疫细胞亚群的差异。通过PD-1免疫检查点抑制剂治疗反应分析、外部数据集验证以及不同药物的半数抑制浓度(IC50)值,研究高风险组与低风险组在药物治疗效果上可能存在的差异。④体外培养PIG1(正常人黑色素细胞系)和A375(人黑色素瘤细胞系)细胞,通过Western Blot 和RT-qPCR检测该预后模型中基因在黑色素细胞和人黑色素瘤细胞中的蛋白表达及mRNA相对表达差异。
结果
2
①分析TCGA和GTEx数据库发现,除
PARK14
(
PLA2G6
)外,所有帕金森病家族基因在黑色素瘤组织和正常组织中的表达均存在显著差异。分析HPA数据库发现,与正常组织相比,帕金森家族基因在黑色素瘤样本中蛋白表达发生上调或下调。在GEPIA2数据库分析帕金森家族基因在不同TNM分期及Clark分级的表达,均发现部分基因表达存在差异。②基于TCGA数据库选择COX回归分析发现其中13个基因表达水平可预测黑色素瘤患者总生存期,对该数据 LASSO回归分析构建包含7个帕金森家族基因(
LRRK2、UCHL1、SNCA、VPS13C、SPR、EIF4G1、FBXO7
)的黑色素瘤预后模型。采用风险因子列联图计算所有患者预后风险评分,将患者分为高、低风险组(中位数=-0.145),结果显示高风险组生存率显著较差,而风险评分越低,生存时间越长。随后对该模型在TCGA队列中进行内部验证:3年和5年的列线图校准曲线显示,黑色素瘤患者预后的预测曲线与实际曲线非常接近。在GEO队列中进行外部验证:在GEO数据库中选择3个包含完整临床数据和基因表达矩阵的数据集(GSE19234、GSE22153 和 GSE53118),基于预后模型的风险评分在ROC曲线上的敏感性和特异性优于TNM分期。GO富集和KEGG通路富集发现7个基因主要富集在肿瘤发生和肿瘤免疫的通路中,如磷酸化、程序性细胞死亡、细胞周期等,这表明预后模型与肿瘤发生和免疫相关。③GSVA评分发现,这7个基因在与肿瘤发生和抗肿瘤免疫相关的通路中富集,能抑制肿瘤生长的CD8
+
T细胞、细胞毒性细胞、Th1细胞则在低风险组中浸润更多。同时,低风险组患者肿瘤细胞对尼洛替尼(nilotinib)、BIBW2992、博来霉素(bleomycin)、硼替佐米(bortezomib)、达沙替尼(dasatinib)和MK-2206的IC50值较低(
P
<
0.05)。④通过RT-qPCR和Western blot分析,以在转录和翻译水平上验证A375(人恶性黑色素瘤细胞系)和PIG1细胞(正常皮肤黑色素细胞系)中这七个基因的相对表达差异。实验结果显示,
LRRK2
和
UCHL1
基因在A375细胞和PIG1细胞中几乎不表达,同时,与正常黑色素细胞相比,黑色素瘤细胞中
SNCA、VPS13C、SPR、EIF4G1
和
FBXO7
基因的mRNA表达显著上调(
P
<
0.05);SPR、eIF4G1和FBXO7的蛋白表达显著上调(
P
<
0.05)。
结论
2
本研究建立的七基因特征预后模型能够有效评估CMM患者预后及其药物治疗疗效。
Objective
2
To explore the genetic characteristics of familial Parkinson’s disease (PD)-related genes and construct a model of the gene signatures to guide the prognosis and treatment of patients with skin cutaneous melanoma (CMM).
Methods
2
① By analyzing the TCGA and GTEx databases
the expression differences of familial PD-related genes were compared between melanoma and normal tissues. The GEPIA2 database was used to analyze the expression differences of related genes across different TNM stages and Clark levels. ② Based on the TCGA database
a signature model containing 7 key genes was established using prognostic forest plot
LASSO regression model
and risk factor analysis. Internal and external validation of this model was performed
and GO enrichment and KEGG pathway enrichment analyses were conducted for these genes. ③ Based on the TIMER database
the impacts of gene mutations in the prognostic model on immune cell infiltration in the tumor microenvironment (TME) were analyzed. ssGSEA heatmaps were employed to analyze differences in immune cell subsets between high-risk and low-risk groups. Through PD-1 immune checkpoint inhibitor treatment response analysis
external dataset validation
and half-maximal inhibitory concentration (IC50) values of different drugs
possible differences in drug treatment efficacy between high-risk and low-risk groups were investigated. ④ Western blotting and RT-qPCR were utilized to detect the protein expression and relative mRNA expression differences of the genes in this prognostic model between normal human melanocyte cell line PIG1 and human melanoma cell line A375.
Results
2
① Analysis of the TCGA and GTEx databases revealed that
except for
PARK14
(
PLA2G6
)
all familial PD-related genes showed significant expression differences between melanoma tissues and normal tissues. Analysis of the HPA database found that the protein expression of familial PD-related genes was either up-regulated or down-regulated in melanoma samples compared to normal tissues. Analysis of the GEPIA2 database for the expression of these genes across different TNM stages and Clark levels also found differential expression in some genes. ② COX regression analysis based on TCGA database identified that expression levels of 13 genes could predict overall survival in melanoma patients. LASSO regression analysis of this data constructed a melanoma prognostic model containin
g 7 familial PD-related genes (
LRRK2
UCHL1
SNCA
VPS13C
SPR
EIF4G1
FBXO7
). The prognostic risk scores for all patients were calculated using a risk factor contingency plot
and then the patients were divided into high-risk and low-risk groups based on the median risk score (-0.145). The results showed that the high-risk group had a significantly worse survival rate
while the low-risk group exhibited a longer survival time. Internal validation of the model in the TCGA cohort: the calibration curves for the 3-year and 5-year nomograms showed that the predicted curves for melanoma patient prognosis were very close to the actual curves. External validation in GEO cohorts: 3 datasets from the GEO database containing complete clinical data and gene expression matrices (GSE19234
GSE22153
and GSE53118) were selected. Based on the prognostic model's risk score
the sensitivity and specificity on the ROC curve were better than TNM staging. GO and KEGG pathway enrichment analyses found that the 7 genes were mainly enriched in pathways related to tumorigenesis and tumor immunity
such as phosphorylation
programmed cell death
and cell cycle
suggesting the prognostic model is associated with tumorigenesis and immunity. ③ GSVA scoring revealed that these 7 genes were enriched in pathways related to tumorigenesis and anti-tumor immunity. CD8
+
T cells
cytotoxic cells
and Th1 cells
which can inhibit tumor growth
were more infiltrated in the low-risk group. Meanwhile
the IC
50
values for nilotinib
BIBW2992
bleomycin
bortezomib
dasatinib
and MK-2206 were lower in tumor cells from low-risk group patients (
P
<
0.05). ④ RT-qPCR and Western blotting analysis were used to verify the relative expression differences of these 7 genes at the transcriptional and translational levels in A375 and PIG1 cells. The experimental results showed that
LRRK2
a
nd
UCHL1
were almost not expressed in both A375 and PIG1 cells. Meanwhile
compared to normal melanocytes
the mRNA expression of
SNCA
VPS13C
SPR
eIF4G1
and
FBXO7
genes was significantly up-regulated in melanoma cells (
P
<
0.05). The protein expression of SPR
EIF4G1
and FBXO7 was significantly upregulated (
P
<
0.05).
Conclusion
2
The 7-gene signature prognostic model developed in this study can effectively assess prognosis and treatment efficacy in CMM patients.
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