半胱氨酸酶抑制剂A在口腔鳞癌中的表达及临床意义

doi:10.19438/j.cjoms.2021.06.003

中国口腔颌面外科杂志 ›› 2021, Vol. 19 ›› Issue (6): 494-499.doi: 10.19438/j.cjoms.2021.06.003

半胱氨酸酶抑制剂A在口腔鳞癌中的表达及临床意义

杨云博^1,2,*, 王钰璞^2,*, 李行^1,2, 韩楠男², 严明², 阮敏²

1. 潍坊医学院口腔医学院,山东潍坊 261053;
2. 上海交通大学医学院附属第九人民医院口腔颌面-头颈肿瘤科,上海交通大学口腔医学院,国家口腔医学中心,国家口腔疾病临床医学研究中心,上海市口腔医学重点实验室,上海 200011

出版日期:2021-12-25 发布日期:2022-03-02
通讯作者: 阮敏,E-mail：doctorruanmin@sjtu.edu.cn
作者简介:杨云博（1994-），男，在读硕士研究生，E-mail：565350445@qq.com；王钰璞（1995-），男，住院医师，E-mail：wypf4@vip.qq.com。*并列第一作者
基金资助:
国家自然科学基金（82072983、81777270）; 上海市卫生健康委员会研究基金（201740186）; 上海市奉贤区临床诊疗中心建设项目[（2017）118]; 上海市奉贤区科学技术委员会科学技术发展基金（20170702）

Expression and clinical significance of cystatin A in oral squamous cell carcinoma

YANG Yun-bo^1,2, WANG Yu-pu¹, LI Hang^1,2, HAN Nan-nan², YAN Ming², RUAN Min²

1. School of Stomatology, Weifang Medical University. Weifang 261053, Shandong Province;
2. Department of Oromaxillofacial Head and Neck Oncology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology. Shanghai 200011, China

Online:2021-12-25 Published:2022-03-02

摘要/Abstract

摘要： 目的探讨半胱氨酸酶抑制剂A（CSTA）在口腔鳞癌中的表达及其与临床病理特征的关系。方法通过免疫组织化学染色、Western 免疫印迹和实时定量PCR（qRT-PCR）检测CSTA的表达,通过Transwell迁移侵袭实验评估CSTA对口腔鳞癌细胞转移的促进作用。采用SPSS 25.0和GraphPad Prism 8.4.3软件进行数据分析。结果 CSTA在口腔鳞癌中的表达显著低于正常口腔黏膜上皮组织,其表达量与颈淋巴结转移（P=0.028）和肿瘤病理级别呈显著负相关（P=0.001）,CSTA低表达的细胞倾向分布于肿瘤侵袭前缘;CSTA低表达组患者总生存期显著短于CSTA高表达组,CSTA过表达可在体外抑制口腔鳞癌细胞迁移与侵袭。结论 CSTA在调控口腔鳞癌颈淋巴转移过程中发挥重要作用,有望成为口腔鳞癌颈淋巴转移的潜在预测和治疗靶标。

关键词: 半胱氨酸蛋白酶抑制剂A, 口腔鳞癌, 颈淋巴转移, 肿瘤浸润, 生物标记

Abstract: PURPOSE: To evaluate the expression of cystatin A (CSTA) in oral squamous cell carcinoma (OSCC), and its association with clinicopathological factors. METHODS: The expression of CSTA was examined using quantitative real-time PCR, immunohistochemistry, and Western blotting. Statistical analysis was performed to determine the association between CSTA expression and clinicopathological characteristics of patients with OSCC. The effect of CSTA on migration and invasion was evaluated using in vitro assays. The results were analyzed using SPSS 25.0 and GraphPad Prism 8.4.3 software. RESULTS: CSTA expression was significantly downregulated in primary OSCC tissues, and lower CSTA expression was significantly correlated with high tumor grade (P=0.001), cervical lymph node metastasis(P=0.028), and poor overall survival(P<0.001). CTSA overexpression inhibited OSCC cell migration and invasion in vitro. CONCLUSIONS: This study revealed that CSTA is a promising biomarker and therapeutic target with prognostic implications in patients with OSCC. CSTA may play an essential role in cervical lymph node metastasis and tumor differentiation.

Key words: Cystatin A, Oral squamous cell carcinoma, Cervical lymphatic metastasis, Neoplasm invasiveness, Biomarker

中图分类号:

R739.8

杨云博, 王钰璞, 李行, 韩楠男, 严明, 阮敏. 半胱氨酸酶抑制剂A在口腔鳞癌中的表达及临床意义[J]. 中国口腔颌面外科杂志, 2021, 19(6): 494-499.

YANG Yun-bo, WANG Yu-pu, LI Hang, HAN Nan-nan, YAN Ming, RUAN Min. Expression and clinical significance of cystatin A in oral squamous cell carcinoma[J]. China J Oral Maxillofac Surg, 2021, 19(6): 494-499.

参考文献

[1] Bray F, Ferlay J, Soerjomataram I, et al.Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin, 2018, 68(6): 394-424.
[2] Siegel RL, Miller KD, Jemal A.Cancer statistics, 2020[J]. CA Cancer J Clin, 2020, 70(1): 7-30.
[3] Sasahira T, Kurihara M, Bhawal UK, et al.Downregulation of miR-126 induces angiogenesis and lymphangiogenesis by activation of VEGF-A in oral cancer[J]. Br J Cancer, 2012, 107(4): 700-706.
[4] Chi AC, Day TA, Neville BW.Oral cavity and oropharyngeal squamous cell carcinoma-an update[J]. CA Cancer J Clin, 2015, 65(5): 401-421.
[5] Ho AS, Kim S, Tighiouart M, et al.Metastatic lymph node burden and survival in oral cavity cancer[J]. J Clin Oncol, 2017, 35(31): 3601-3609.
[6] O'Donnell RK, Kupferman M, Wei SJ, et al. Gene expression signature predicts lymphatic metastasis in squamous cell carcinoma of the oral cavity[J]. Oncogene, 2005, 24(7): 1244-1251.
[7] Cerezo L, Millán I, Torre A, et al.Prognostic factors for survival and tumor control in cervical lymph node metastases from head and neck cancer. A multivariate study of 492 cases[J]. Cancer, 1992, 69(5): 1224-1234.
[8] Jenko S, Dolenc I, Guncar G, et al.Crystal structure of Stefin A in complex with cathepsin H: N-terminal residues of inhibitors can adapt to the active sites of endo- and exopeptidases[J]. J Mol Biol, 2003, 326(3): 875-885.
[9] Pavlova A, Björk I.The role of the second binding loop of the cysteine protease inhibitor, cystatin A (stefin A), in stabilizing complexes with target proteases is exerted predominantly by Leu73[J]. Eur J Biochem, 2002, 269(22): 5649-5658.
[10] Renko M, Požgan U, Majera D, et al.Stefin A displaces the occluding loop of cathepsin B only by as much as required to bind to the active site cleft[J]. FEBS J, 2010, 277(20): 4338-4345.
[11] Blaydon DC, Nitoiu D, Eckl KM, et al.Mutations in CSTA, encoding Cystatin A, underlie exfoliative ichthyosis and reveal a role for this protease inhibitor in cell-cell adhesion[J]. Am J Hum Genet, 2011, 89(4): 564-571.
[12] Pernu H, Räsänen O, Salo T, et al.Cystatin A and B in the development of human squamous epithelia[J]. Acta Histochem, 1990, 88(1): 53-57.
[13] Roesch-Ely M, Nees M, Karsai S, et al.Proteomic analysis reveals successive aberrations in protein expression from healthy mucosa to invasive head and neck cancer[J]. Oncogene, 2007, 26(1): 54-64.
[14] Sinha AA, Quast BJ, Wilson MJ, et al.Prediction of pelvic lymph node metastasis by the ratio of cathepsin B to stefin A in patients with prostate carcinoma[J]. Cancer, 2002, 94(12): 3141-3149.
[15] Lin YY, Chen ZW, Lin ZP, et al.Tissue levels of stefin a and stefin b in hepatocellular carcinoma[J]. Anat Rec (Hoboken), 2016, 299(4): 428-438.
[16] Li C, Chen L, Wang J, et al.Expression and clinical significance of cathepsin B and stefin A in laryngeal cancer[J]. Oncol Rep, 2011, 26(4): 869-875.
[17] Kowalewska M, Radziszewski J, Goryca K, et al.Estimation of groin recurrence risk in patients with squamous cell vulvar carcinoma by the assessment of marker gene expression in the lymph nodes[J]. BMC Cancer, 2012, 12: 223-233.
[18] Puram SV, Tirosh I, Parikh AS, et al. Single-cell transcriptomic analysis of primary and metastatic tumor ecosystems in head and neck cancer [J]. Cell, 2017, 171(7): 1611-24.e24.
[19] Aiello NM, Kang Y.Context-dependent EMT programs in cancer metastasis[J]. J Exp Med, 2019, 216(5): 1016-1026.
[20] Parikh AS, Puram SV, Faquin WC, et al.Immunohistochemical quantification of partial-EMT in oral cavity squamous cell carcinoma primary tumors is associated with nodal metastasis[J]. Oral Oncol, 2019, 99: 104458.
[21] Su B, Gao L, Baranowski C, et al.A genome-wide RNAi screen identifies FOXO4 as a metastasis-suppressor through counteracting PI3K/AKT signal pathway in prostate cancer[J]. PLoS One, 2014, 9(7): e101411.
[22] Dong T, Zhang Y, Chen Y, et al.FOXO1 inhibits the invasion and metastasis of hepatocellular carcinoma by reversing ZEB2-induced epithelial-mesenchymal transition[J]. Oncotarget, 2017, 8(1): 1703-1713.
[23] Hsu MC, Pan MR, Hung WC.Two birds, one stone: double hits on tumor growth and lymphangiogenesis by targeting vascular endothelial growth factor receptor 3[J]. Cells, 2019, 8(3): 270-290.
[24] Strojan P, Budihna M, Smid L, et al.Prognostic significance of cysteine proteinases cathepsins B and L and their endogenous inhibitors stefins A and B in patients with squamous cell carcinoma of the head and neck[J]. Clin Cancer Res, 2000, 6(3): 1052-1062.
[25] Aniĉin A, Gale N, Smid L, et al.Expression of stefin A is of prognostic significance in squamous cell carcinoma of the head and neck[J]. Eur Arch Otorhinolaryngol, 2013, 270(12): 3143-3151.
[26] Flezar MS, Kirbis IS, Popovic KS, et al.Radiosensitivity of squamous cell carcinoma metastases to the neck assessed by immunocytochemical profiling of fine-needle aspiration biopsy cell specimens: a pilot study[J]. Radiother Oncol, 2009, 93(3): 575-580.
[27] Muttardi K, Nitoiu D, Kelsell DP, et al.Acral peeling skin syndrome associated with a novel CSTA gene mutation[J]. Clin Exp Dermatol, 2016, 41(4): 394-398.
[28] Krunic AL, Stone KL, Simpson MA, et al.Acral peeling skin syndrome resulting from a homozygous nonsense mutation in the CSTA gene encoding cystatin A[J]. Pediatr Dermatol, 2013, 30(5): e87-e88.
[29] Gupta A, Nitoiu D, Brennan-Crispi D, et al.Cell cycle- and cancer-associated gene networks activated by Dsg2: evidence of cystatin A deregulation and a potential role in cell-cell adhesion[J]. PLoS One, 2015, 10(3): e0120091.
[30] Carnielli CM, Macedo CCS, De Rossi T, et al.Combining discovery and targeted proteomics reveals a prognostic signature in oral cancer[J]. Nat Commun, 2018, 9(1): 3598.
[31] Oler G, Camacho CP, Hojaij FC, et al.Gene expression profiling of papillary thyroid carcinoma identifies transcripts correlated with BRAF mutational status and lymph node metastasis[J]. Clin Cancer Res, 2008, 14(15): 4735-4742.
[32] Vigneswaran N, Wu J, Zacharias W.Upregulation of cystatin M during the progression of oropharyngeal squamous cell carcinoma from primary tumor to metastasis[J]. Oral Oncol, 2003, 39(6): 559-568.

半胱氨酸酶抑制剂A在口腔鳞癌中的表达及临床意义

Expression and clinical significance of cystatin A in oral squamous cell carcinoma

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