AR、SKP2、SOX10、PD-L1及TIL表达在三阴性乳腺癌中的意义

doi:10.16150/j.1671-2870.2024.02.010

摘要/Abstract

摘要：

目的：探索雄激素受体(androgen receptor，AR)、S期激酶相关蛋白2（S-phase kinase-associated protein 2，SKP2）、性别决定区Y相关的HMG盒含因子10（sry-related HMG box-containing factor 10，SOX10）、程序性死亡配体1（programmed death-ligand 1，PD-L1）及肿瘤浸润性淋巴细胞（tumor infiltrating lymphocyte，TIL）在三阴性乳腺癌（triple negative breast cancer，TNBC）表达与临床病理特征和预后的关系。方法：根据苏木精-伊红染色（hematoxylin-eosin, HE）染色切片评判109例TNBC瘤巢内TIL的比例，采用Leica Bond-Max全自动免疫组化仪检测TNBC组织中AR、SKP2、SOX10、PD-L1的表达。分析以上各生物指标与临床病理特征间的关系，并采用kaplan-Meier、Log-rank进行生存分析。结果：95例患者获得随访，中位随访时间为48个月，中位无病生存时间（disease-free survival, DFS）为42个月，中位总生存时间（overall survival, OS）48个月。在TNBC中，AR阳性表达与淋巴结转移阴性（P=0.009）、肿瘤最大径<2 cm（P=0.008）相关，TIL高表达与低级别TNBC相关（P=0.007），SKP2阳性表达与神经/脉管侵犯阳性（P=0.011）、高级别TNBC相关（P=0.002），SOX10阳性表达与淋巴结转移阳性（P=0.022）、高级别TNBC（P=0.005）相关，PD-L1阳性表达与淋巴结转移阳性（P=0.020）、神经/脉管侵犯阳性（P=0.006）、高级别TNBC（P=0.042）相关。生存分析显示，SKP2、SOX10阳性表达与更差的DFS（P=0.007、P<0.001）和OS（P=0.013、P<0.001）相关，TIL高表达与更好的DFS（P=0.016）及OS（P=0.004）相关。在生物表志物的联合表达中，AR+/SKP2-、AR+/SOX10-与更好的DFS（P=0.004、P<0.001）及OS（P=0.007、P=0.001）相关，SOX10+/低TIL、PD-L1+/低TIL与更差的DFS（P<0.001、P=0.008）及OS（P=0.001、P=0.002）相关，AR-/低TIL者具有更差的OS（P=0.014）。SKP2（HR=4.143，95%CI为1.578~10.875）、SOX10（HR=7.578，95%CI为2.067~27.782）的阳性表达是影响TNBC患者DFS的独立预后因子，SKP2（HR=3.758，95%CI为1.400~10.084）、SOX10（HR=5.131，95%CI为1.316~20.000）及TIL（HR=0.375，95%CI为0.154~0.917）的阳性表达是TNBC患者OS的独立预后因子（P均<0.05）。结论：在TNBC中，AR阳性、TIL高表达与具有更好预后的临床病理特征相关，SKP2、SOX10和PD-L1与具侵袭性的临床病理特征相关。SKP2、SOX10及TIL表达与TNBC预后相关，提示这些生物指标可能成为TNBC新的预后因子，同时它们也有可能成为潜在的治疗靶点。

关键词: 三阴性乳腺癌, 雄激素受体, S期激酶相关蛋白2, 性别决定区Y相关的HMG盒含因子10, 程序性死亡配体1, 肿瘤浸润性淋巴细胞

Abstract:

Objective To explore the expression of androgen receptor (AR), S-phase kinase associated protein 2 (SKP2), Sry-related HMG box-containing factor 10 (SOX10), programmed death-ligand 1 (PD-L1) and tumor infiltrating lymphocytes (TILs) in triple-negative breast cancer (TNBC) and their relationships with clinical prognosis. Methods The proportion of TILs in 109 TNBCs was assessed on Hematoxylin-eosin stained sections ，and Leica Bond-Max automatic immunohistochemistry apparatus was used to detect the expressions of AR,SKP2,SOX10 and PD-L1 in TNBC tissue. The relationship between the above indicators and clinicopathological charactersitics was analyzed. Univariate survival analysis was performed by Kaplan-Meier, and survival by Log rank test. Multivariate survival analysis was performed by cox regression model. Results A total of 95 patients were followed-up with a median follow-up time of 48 months. For 95patients,the median disease-free survival (DFS) time was 42 months, and median overall survival (OS) time were 48 months. In TNBC, the expression of AR was associated with negative lymph node metastasis, maximum tumor diameter <2 cm. High expression of TILs was associated with low grade TNBC. The expression of SKP2 was associated with positive nerve/vasculature invasion and high grade TNBC. The expression of SOX10 was associated with high grade TNBC and positive lymph node metastasis. The expression of PD-L1 was associated with positive lymph node metastasis, positive nerve/vascular invasion, and high grade TNBC (all the P<0.05 as above). Survival analysis demonstrated that the positive expression of SKP2 or AR SOX10 was correlated with worse DFS (P=0.007、P<0.001) and OS (P=0.013、P<0.001), and patients with high expression of TILs showed better DFS (P=0.016) and OS (P=0.004). TNBC patients with AR+/SKP2- or AR+/SOX10- had better DFS (P=0.004、P<0.001) and OS (P=0.007、P<0.001), while those with SOX10+/low TILs or PD-L1+/low TILs had worse DFS (P=0.000、P=0.008) and OS (P=0.001、P=0.002), and AR-/low TILs had worse OS (P=0.014). Multivariate survival analysis showed positive expression SKP2 (HR=4.143, 95%CI 1.578-10.875), or SOX10 (HR=7.578, 95%CI 2.067-27.782) were independent prognostic factors for worse DFS, postive expression SKP2 (HR=3.758, 95%CI 1.400-10.084), or SOX10 (HR=5.131, 95%CI 1.316-20.000)， were independent prognostic factors for worse OS (all the P<0.05 as above) and higher TILs(HR=0.375,95%CI 0.154-0.917) was independent prognostic factors for better OS(all the P<0.05 as above). Conclusions High expression of AR and TILs in TNBC indicate better prognosis, while SKP2, SOX10 and PD-L1 expression demonstrate more aggressive clinicopathological features in TNBC. The expression of SKP2, SOX10 and TILs in TNBC are associated with prognosis, suggesting that these indicators may serve as new prognostic factors and potential therapeutic targets in TNBC.

Key words: Triple-negative breast cancer, Androgen receptor, S-phase kinase associated protein 2, Sry-related HMG box-containing factor 10, Programmed death-ligand 1, Tumor infiltrating lymphocytes

中图分类号:

刘娟, 殷丽娟, 范德生. AR、SKP2、SOX10、PD-L1及TIL表达在三阴性乳腺癌中的意义[J]. 诊断学理论与实践, 2024, 23(02): 162-172.

LIU Juan, YIN Lijuan, FAN Desheng. The clinicopathologic significance of AR, SKP2, SOX10, PD-L1 and TILs expression in triple-negative breast cancer[J]. Journal of Diagnostics Concepts & Practice, 2024, 23(02): 162-172.

图/表 9

图1

表1

表2

表3

图2

图3

图4

图5

表4

参考文献 27

[1]	CHOUPANI E, MAHMOUDI GOMARI M, ZANGANEH S, et al. Newly developed targeted therapies against the androgen receptor in triple-negative breast cancer: a review[J]. Pharmacol Rev, 2023, 75(2):309-327. doi: 10.1124/pharmrev.122.000665 pmid: 36781219
[2]	NITĂ I, NITIPIR C, TOMAS A, et al. Correlation between androgen receptor expression and immunohistochemistry type as prognostic factors in a cohort of breast cancer patients: result from a single-center, cross sectional study[J]. Healthcare (Basel), 2021, 9(3):277.
[3]	GOTO Y, THIKE A A, ONG C C H, et al. Characteristics, behaviour and role of biomarkers in metastatic triple-negative breast cancer[J]. J Clin Pathol, 2020, 73(3):147-153. doi: 10.1136/jclinpath-2019-206078 pmid: 31563883
[4]	SANG M, MENG L, MA C, et al. Effect of AR antagonist combined with PARP1 inhibitor on sporadic triple-negative breast cancer bearing AR expression and methylation-mediated BRCA1 dysfunction[J]. Biomed Pharmacother, 2019, 111:169-177. doi: S0753-3322(18)37264-0 pmid: 30580238
[5]	SRIDHAR N, GLISCH C, JAWA Z, et al. Androgen receptor expression in patients with triple negative breast cancer treated with neoadjuvant chemotherapy: a single institution study[J]. J Cancer, 2022, 13(8):2472-2476. doi: 10.7150/jca.67536 pmid: 35711833
[6]	段帅, 地力木拉提·艾斯木吐拉, 王海燕, 等. 三阴性乳腺癌新辅助化疗病理完全缓解的影响因素[J]. 中国临床研究, 2024, 37(3):354-358.
	DUAN S, DILIMULATI A, WANG H Y, et al. Influen-cing factors of pathological complete response in triple-negative breastcancer with neoadiuvant chemotherapy[J]. Chin J Clin Res, 2024, 37(3):354-358.
[7]	DAI C, ELLISEN L W. Revisiting androgen receptor signaling in breast cancer[J]. Oncologist, 2023, 28(5):383-391.
[8]	LI C, DU L, REN Y, et al. SKP2 promotes breast cancer tumorigenesis and radiation tolerance through PDCD4 ubiquitination[J]. J Exp Clin Cancer Res, 2019, 38(1):76. doi: 10.1186/s13046-019-1069-3 pmid: 30760284
[9]	WANG X, ZHANG T, ZHANG S, et al. Prognostic values of F-box members in breast cancer: an online database analysis and literature review[J]. Biosci Rep, 2019, 39(1):BSR20180949.
[10]	SAUNUS J M, DE LUCA X M, NORTHWOOD K, et al. Epigenome erosion and SOX10 drive neural crest phenotypic mimicry in triple-negative breast cancer[J]. NPJ Breast Cancer, 2022, 8(1):57. doi: 10.1038/s41523-022-00425-x pmid: 35501337
[11]	ALI S, RATHORE Z, RAFIQUE Z, et al. Expression of SOX10 in triple-negative breast carcinoma in Pakistan[J]. Cureus, 2022, 14(8):e27938.
[12]	JIN L, QIN C, QI X, et al. Clinicopathological significance of Sox10 expression in triple-negative breast carcinoma[J]. Transl Cancer Res, 2020, 9(9):5603-5613. doi: 10.21037/tcr-20-2634 pmid: 35117924
[13]	KRIEGSMANN K, FLECHTENMACHER C, HEIL J, et al. Immunohistological expression of SOX-10 in triple-negative breast cancer: a descriptive analysis of 113 samples[J]. Int J Mol Sci, 2020, 21(17):6407.
[14]	PARVATHAREDDY S K, SIRAJ A K, AHMED S O, et al. PD-L1 protein expression in middle eastern breast cancer predicts favorable outcome in triple-negative breast cancer[J]. Cells, 2021, 10(2):229.
[15]	CHOI S H, CHANG J S, KOO J S, et al. Differential prognostic impact of strong PD-L1 expression and ¹⁸F-FDG uptake in triple-negative breast cancer[J]. Am J Clin Oncol, 2018, 41(11):1049-1057.
[16]	SUNAR V, T DOGAN H, SARICI F, et al. Association between androgen receptor status and prognosis in triple negative breast cancer[J]. J BUON, 2018, 23(5):1325-1330. pmid: 30570854
[17]	ZHONG K, YANG F, HAN Q, et al. Skp2 expression has different clinicopathological and prognostic implications in lung adenocarcinoma and squamous cell carcinoma[J]. Oncol Lett, 2018, 16(3):2873-2880. doi: 10.3892/ol.2018.9000 pmid: 30127874
[18]	SHI Y, LI N, REN H, et al. Prognostic role of s-phase kinase-associated protein 2 in breast cancer: A meta-analysis[J]. Indian J Cancer, 2015, 52 Suppl 3:E153-E157.
[19]	FENG W, LIU S, ZHU R, et al. SOX10 induced Nestin expression regulates cancer stem cell properties of TNBC cells[J]. Biochem Biophys Res Commun, 2017, 485(2):522-528.
[20]	RAMMAL R, GOEL K, ELISHAEV E, et al. The utility of SOX10 immunohistochemical staining in breast patho-logy[J]. Am J Clin Pathol, 2022, 158(5):616-625.
[21]	刘建兰, 陈黛诗, 成志强, 等. SOX10和GATA3在乳腺癌中的表达及意义[J]. 中华病理学杂志, 2022, 51(6):536-541.
	LIU J L, CHEN D S, CHENG Z Q, et al. Expression of SOX10 and GATA3 in breast cancer and their significance[J]. Chin J Pathol, 2022, 51(6):536-541.
[22]	LI Y, OPYRCHAL M, YAO S, et al. The role of programmed death ligand-1 and tumor-infiltrating lymphocytes in breast cancer overexpressing HER2 gene[J]. Breast Cancer Res Treat, 2018, 170(2):293-302.
[23]	MANGIA A, SAPONARO C, VAGHEGGINI A, et al. Should tumor infiltrating lymphocytes, androgen receptor, and FOXA1 expression predict the clinical outcome in triple negative breast cancer patients?[J]. Cancers (Basel), 2019, 11(9):1393.
[24]	TOMIOKA N, HATANAKA K C, OKUYAMA D, et al. Programmed death ligand 1-positive immune cells in primary tumor or metastatic axillary lymph nodes can predict prognosis of triple-negative breast cancer even when present at <1% in the tumor region[J]. Breast Cancer, 2023, 30(3):497-505.
[25]	YAZAKI S, SHIMOI T, YOSHIDA M, et al. Integrative prognostic analysis of tumor-infiltrating lymphocytes, CD8, CD20, programmed cell death-ligand 1, and tertiary lymphoid structures in patients with early-stage triple-negative breast cancer who did not receive adjuvant chemotherapy[J]. Breast Cancer Res Treat, 2023, 197(2):287-297.
[26]	MORI H, KUBO M, YAMAGUCHI R, et al. The combination of PD-L1 expression and decreased tumor-infiltra-ting lymphocytes is associated with a poor prognosis in triple-negative breast cancer[J]. Oncotarget, 2017, 8(9):15584-15592.
[27]	袁杰, 张华, 王群, 等. 影响乳腺癌新辅助化疗后前哨淋巴结活检准确率的因素[J]. 中国临床研究, 2023, 36(11):1683-1687.
	YUAN J, ZHANG H, WANG Q, et al. Factors influen-cing the accuracy of sentinel lymph node biopsy after neoadjuvant chemotherapy in breast cancer[J]. Chin J Clin Res, 2023, 36(11):1683-1687.

clinicopathological features	n	AR positive (%)	χ²	P	SKP2 positive (%)	χ²	P	SOX10 positive (%)	χ²	P
Age (year)			1.195	0.274		0.006	0.940		1.330	0.249
<50	36	6(16.7)			16(44.4)			17(47.2)
≥50	73	19(26.0)			33(45.2)			43(58.9)
Maximum tumor diameter (cm)			6.960	0.008		0.097	0.755		0.614	0.433
<2	23	10(43.5)			11(47.8)			11(47.8)
≥2	86	15(17.4)			38(44.2)			49(57.0)
Histological grade			2.484	0.115		9.722	0.002		7.934	0.005
Low	42	13(31.0)			11(26.2)			16(38.1)
High	67	12(17.9)			38(56.7)			44(65.7)
Lymph node metastasis			6.767	0.009		0.172	0.679		5.231	0.022
Negative	58	19(32.8)			25(43.1)			26(44.8)
Positive	51	6(11.8)			24(47.1)			34(66.7)
Nerve/vascular invasion			1.319	0.251		6.478	0.011		1.177	0.278
Negative	91	19(20.9)			36(39.6)			48(52.7)
Positive	18	6(33.3)			13(72.2)			12(66.7)

Clinicopathological features	n	PD-L1 positive (%)	χ²	P	low TIL (%)	moderate TIL (%)	high TIL (%)	χ²	P
Age (year)			0.392	0.531				1.127	0.569
<50	36	14(38.9)			15(41.7)	16(44.4)	5(13.9)
≥50	73	33(45.2)			24(32.9)	34(46.6)	15(20.5)
Maximum tumor diameter (cm)			0.826	0.363				1.401	0.496
<2	23	8(34.8)			7(30.4)	13(56.5)	3(13.0)
≥2	86	39(45.3)			32(37.2)	37(43.0)	17(19.8)
Histological grade			4.124	0.042				9.894	0.007
Low	42	13(31.0)			9(21.4)	20(47.6)	13(31.0)
High	67	34(50.7)			30(44.8)	30(44.8)	7(10.4)
Lymph node metastasis			5.425	0.020				2.281	0.320
Negative	58	19(32.8)			17(29.3)	29(50.0)	12(20.7)
Positive	51	28(54.9)			22(43.1)	21(41.2)	8(15.7)
Nerve/vascular invasion			7.446	0.006				0.715	0.699
Negative	91	34(37.4)			31(34.1)	43(47.3)	17(18.7)
Positive	18	13(72.2)			8(44.4)	7(38.9)	3(16.7)

Clinicopathological features	Patients n=95	Patients with relapse and metastasis n=31(%)	Dead patients n=26(%)	DFS		OS
Clinicopathological features	Patients n=95	Patients with relapse and metastasis n=31(%)	Dead patients n=26(%)	χ²	P	χ²	P
Age (year)				1.162	0.281	0.273	0.601
<50	29	12(41.4)	9(31.0)
≥50	66	19(28.8)	17(25.8)
Maximum tumor diameter (cm)				0.490	0.484	0.535	0.465
<2	18	5(27.8)	4(22.2)
≥2	77	26(33.8)	22(28.6)
Histological grade				3.285	0.070	2.714	0.099
Low	36	9(25.0)	8(22.2)
High	59	22(37.3)	18(30.5)
Lymph node metastasis				9.303	0.002	7.138	0.008
Negative	50	10(20.0)	9(18.0)
Positive	45	21(46.7)	17(37.8)
Nerve/vascular invasion				2.531	0,112	1.173	0.279
Negative	80	24(30.0)	21(26.2)
Positive	15	7(46.7)	5(33.3)
Expression of AR				3.845	0.050	3.763	0.052
Negative	72	27(37.5)	23(31.9)
Positive	23	4(17.4)	3(13.0)
Expression of SKP2				7.372	0.007	6.107	0.013
Negative	51	11(21.6)	9(17.6)
Positive	44	20(45.5)	17(38.6)
Expression of SOX10				21.072	0.000	15.777	0.000
Negative	42	3(7.1)	3(7.1)
Positive	53	28(52.8)	23(43.4)
Expression of PD-L1				0.610	0.435	0.238	0.626
Negative	52	15(28.8)	13(25.0)
Positive	43	16(37.2)	13(30.2)
Expression of TIL				8.284	0.016	11.022	0.004
Low	34	16(47.1)	15(44.1)
Moderate	44	12(27.3)	10(22.7)
High	17	3(17.6)	1(5.9)

Covariant	DFS				OS
Covariant	Wald	P	HR	95.0%CI of HR	Wald	P	HR	95.0%CI of HR
Age	3.277	0.070	0.474	0.212_1.064	1.555	0.212	0.569	0.234_1.381
Maximum tumor diameter	0.338	0.561	1.098	0.801_1.504	0.032	0.858	0.968	0.676_1.386
Histological grade	0.578	0.447	0.703	0.284_1.743	0.663	0.415	0.665	0.249_1.777
Lymph node metastasis	4.758	0.029	2.718	1.107_6.676	4.336	0.037	2.896	1.065_7.876
Nerve/vascular invasion	1.741	0.187	0.480	0.161_1.428	1.571	0.210	0.442	0.124_1.583
Expression of AR	0.075	0.784	0.842	0.247_2.876	0.039	0.843	0.866	0.209_3.589
Expression of SKP2	8.334	0.004	4.143	1.578_10.875	6.910	0.009	3.758	1.400_10.084
Expression of SOX10	9.335	0.002	7.578	2.067_27.782	5.550	0.018	5.131	1.316_20.000
Expression of PD-L1	0.015	0.903	1.067	0.376_3.030	0.939	0.333	1.755	0.562_5.478
Expression of TILs	2.015	0.156	0.601	0.298_1.214	4.625	0.032	0.375	0.154_0.917