SHOX2高表达与pT<sub>1</sub>期非透明细胞肾细胞癌淋巴结转移及不良预后的相关性研究

doi:10.3877/cma.j.issn.1674-3253.2026.04.012

目的

探讨SHOX2在pT₁期非透明细胞肾细胞癌（nccRCC）淋巴结转移（LNM）及不良预后中的作用，评估其作为风险分层生物标志物的价值。

方法

回顾性分析单中心528例接受机器人辅助肾部分切除术的pT₁期肾细胞癌（RCC）患者的临床资料，其中96例nccRCC，432例透明细胞肾细胞癌（ccRCC），比较两组的LNM率及预后。从TCGA数据库获取nccRCC转录组数据（n=355），筛选淋巴结阳性（LN+）组与淋巴结阴性（LN-）组间的差异表达基因，并进行生存分析和通路富集分析。对20例pT₁期nccRCC石蜡标本行SHOX2免疫组化染色，比较两组蛋白表达水平。

结果

临床队列中，pT₁期nccRCC的LNM率高于ccRCC（5.21% vs 0.46%，P=0.001），LNM是nccRCC不良预后的独立危险因素（HR=36.44，95%CI：4.67~284.47，P<0.001）。TCGA队列中，SHOX2是LN+组上调最显著的基因（log₂FC=6.01，P<0.001），且SHOX2 mRNA高表达与无病生存率（DFS）缩短相关（HR=2.896，95%CI：1.329~6.310，P=0.007）。功能富集分析提示SHOX2与细胞外基质（ECM）重塑（r=0.401）和上皮间充质转化（EMT）激活（r=0.381）相关（均P<0.001）。nccRCC组织标本初步检测显示，LN+组SHOX2蛋白表达总分高于LN-组（Mann-Whitney U=9.500，P=0.002）。

结论

SHOX2可能通过调控ECM-EMT轴驱动pT₁期nccRCC的淋巴转移，是潜在的预后标志物。未来可探索SHOX2表达联合其甲基化检测在高风险nccRCC中的分层价值，但需多中心前瞻性研究验证。

关键词: 非透明细胞肾细胞癌, SHOX2, 淋巴结转移, 生物标志物, 上皮间充质转化, 细胞外基质

Objective

To investigate the role of SHOX2 in lymph node metastasis (LNM) and poor prognosis of pT1 non-clear cell renal cell carcinoma (nccRCC), and to evaluate its potential as a risk stratification biomarker.

Methods

Clinical data of 528 pT₁ renal cell carcinoma (RCC) patients who underwent robot-assisted partial nephrectomy at a single center were retrospectively analyzed, including 96 patients of nccRCC and 432 patients of clear cell renal cell carcinoma (ccRCC). LNM rates and prognosis were compared between the two groups. Transcriptomic data of nccRCC (n=355) were obtained from the TCGA database to screen differentially expressed genes between positive lymph nodes (LN+) and negative lymph nodes (LN-) groups, followed by survival analysis and pathway enrichment analysis. SHOX2 immunohistochemistry (IHC) was performed on paraffin-embedded specimens from 20 pT₁ nccRCC patients, and protein expression levels were compared between the two groups.

Results

In the clinical cohort, the LNM rate was higher in nccRCC than in ccRCC (5.21% vs 0.46%, P=0.001), and LNM independently predicted poor prognosis (HR=36.44, 95%CI: 4.67-284.47, P<0.001). In the TCGA cohort, SHOX2 was the most upregulated gene in LN+ tumors (log₂FC=6.01, P<0.001), and high SHOX2 mRNA expression was associated with shorter disease-free survival (HR=2.896, 95%CI: 1.329-6.310, P=0.007). Functional enrichment linked SHOX2 to extracellular matrix (ECM) remodeling (r=0.401) and epithelial-mesenchymal transition (EMT) activation (r=0.381) (both P<0.001). Preliminary IHC analysis showed higher SHOX2 protein expression in LN+ than in LN- specimens (Mann-Whitney U=9.500, P=0.002).

Conclusions

SHOX2 may drive lymphatic metastasis in pT₁ nccRCC through the ECM-EMT axis and serve as a potential prognostic biomarker. Future studies could explore the value of combining SHOX2 expression with its methylation detection for risk stratification in high-risk nccRCC, though multicenter prospective validation is required.

Key words: Non-clear cell renal cell carcinoma, SHOX2, Lymph node metastasis, Biomarker, Epithelial-mesenchymal transition, Extracellular matrix

表1 pT₁期非透明细胞肾细胞癌（nccRCC）组与透明细胞肾细胞癌（ccRCC）组患者临床基线资料比较

组别	例数	年龄[岁，(±s)]	体质量指数[kg/m²，(±s)]	肿瘤最大径[cm，(±s)]	性别[例(%)]		结局[例(%)]
组别	例数	年龄[岁，(±s)]	体质量指数[kg/m²，(±s)]	肿瘤最大径[cm，(±s)]	男	女	存活	转移或死亡
nccRCC	96	52.58±14.42	24.95±2.82	3.50±1.44	33 (34.38)	63 (65.62)	90 (93.75)	6 (6.25)
ccRCC	432	55.01±13.23	24.86±3.54	3.71±1.27	165 (38.19)	267 (61.81)	429 (99.31)	3 (0.69)
统计值		t=-1.598	t=0.226	t=-1.376	χ²=0.489		χ²=11.340
P值		0.111	0.821	0.169	0.484		<0.001
组别	淋巴结情况[例(%)]		涉及但未穿透肾包膜[例(%)]		Fuhrman分级[例(%)]
组别	阴性	阳性	阴性	阳性	低级(1和2)	高级(3和4)
nccRCC	91 (94.79)	5 (5.21)	76 (79.17)	20 (20.83)	66 (68.75)	30 (31.25)
ccRCC	430 (99.54)	2 (0.46)	326 (75.46)	106 (24.54)	337 (78.01)	95 (21.99)
统计值	χ²=10.140		χ²=0.540		χ²=3.731
P值	0.001		0.462		0.054

表1 pT₁期非透明细胞肾细胞癌（nccRCC）组与透明细胞肾细胞癌（ccRCC）组患者临床基线资料比较

组别	例数	年龄[岁，(±s)]	体质量指数[kg/m²，(±s)]	肿瘤最大径[cm，(±s)]	性别[例(%)]		结局[例(%)]
组别	例数	年龄[岁，(±s)]	体质量指数[kg/m²，(±s)]	肿瘤最大径[cm，(±s)]	男	女	存活	转移或死亡
nccRCC	96	52.58±14.42	24.95±2.82	3.50±1.44	33 (34.38)	63 (65.62)	90 (93.75)	6 (6.25)
ccRCC	432	55.01±13.23	24.86±3.54	3.71±1.27	165 (38.19)	267 (61.81)	429 (99.31)	3 (0.69)
统计值		t=-1.598	t=0.226	t=-1.376	χ²=0.489		χ²=11.340
P值		0.111	0.821	0.169	0.484		<0.001
组别	淋巴结情况[例(%)]		涉及但未穿透肾包膜[例(%)]		Fuhrman分级[例(%)]
组别	阴性	阳性	阴性	阳性	低级(1和2)	高级(3和4)
nccRCC	91 (94.79)	5 (5.21)	76 (79.17)	20 (20.83)	66 (68.75)	30 (31.25)
ccRCC	430 (99.54)	2 (0.46)	326 (75.46)	106 (24.54)	337 (78.01)	95 (21.99)
统计值	χ²=10.140		χ²=0.540		χ²=3.731
P值	0.001		0.462		0.054

图1 Kaplan-Meier曲线比较pT₁期nccRCC与ccRCC患者5年无病生存率注：阴影区域表示95%置信区间，P<0.001

表2 nccRCC预后不良的单因素和多因素Cox回归分析

因素	单因素Cox分析					多因素Cox分析
因素	β	SE	Z	P	HR (95%CI)	β	SE	Z	P	HR (95%CI)
淋巴结转移阳性	3.65	0.88	4.16	<0.001	38.63 (6.92~215.73)	3.60	1.05	3.43	<0.001	36.44 (4.67~284.47)
涉及但未穿透肾包膜阳性	-0.25	1.10	-0.23	0.821	0.78 (0.09~6.68)	0.39	1.20	0.33	0.742	1.48 (0.14~15.54)
Fuhrman等级高级(3和4)	1.49	0.87	1.72	0.086	4.42 (0.81~24.15)	0.26	1.00	0.26	0.792	1.30 (0.18~9.28)

表2 nccRCC预后不良的单因素和多因素Cox回归分析

因素	单因素Cox分析					多因素Cox分析
因素	β	SE	Z	P	HR (95%CI)	β	SE	Z	P	HR (95%CI)
淋巴结转移阳性	3.65	0.88	4.16	<0.001	38.63 (6.92~215.73)	3.60	1.05	3.43	<0.001	36.44 (4.67~284.47)
涉及但未穿透肾包膜阳性	-0.25	1.10	-0.23	0.821	0.78 (0.09~6.68)	0.39	1.20	0.33	0.742	1.48 (0.14~15.54)
Fuhrman等级高级(3和4)	1.49	0.87	1.72	0.086	4.42 (0.81~24.15)	0.26	1.00	0.26	0.792	1.30 (0.18~9.28)

图2 TCGA数据库筛选LN+与LN-差异表达基因火山图注：SHOX2是淋巴结阳性（LN+）组与淋巴结阴性（LN-）组中上调最显著的基因（log₂FC=6.01，P<0.001）

图3 TCGA来源数据SHOX2高表达与低表达组患者预后分析注：图a为SHOX2表达与生存时间及状态的关系，所有三个子图（上、中、下）的横轴均代表样本，且各子图的样本顺序保持一致；图b为SHOX2高、低表达组的无病生存率Kaplan-Meier曲线；图c为SHOX2预测1、3、5年生存率的时间依赖性ROC曲线

图4 SHOX2基因表达量与肿瘤相关生物学功能的相关性分析注：r为相关系数，Log₂（SHOX2 TPM+1）为SHOX2基因表达量

图5 pT₁期nccRCC癌组织SHOX2免疫组化染色及两组表达总评分比较注：a为不同组织SHOX2蛋白表达水平的免疫组化染色图（×10，×40；标尺为200 μm，50 μm）；b为LN+组与LN-组SHOX2蛋白总评分的比较（Mann-Whitney U=9.500，P=0.002）。

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Association of SHOX2 overexpression with lymph node metastasis and poor prognosis in pT₁ stage non-clear cell renal cell carcinoma

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Association of SHOX2 overexpression with lymph node metastasis and poor prognosis in pT1 stage non-clear cell renal cell carcinoma

Association of SHOX2 overexpression with lymph node metastasis and poor prognosis in pT₁ stage non-clear cell renal cell carcinoma