谷胱甘肽过氧化物酶-3在腮腺多形性腺瘤中的表达及意义

doi:10.3969/j.issn.1005-4979.2020.01.004

摘要/Abstract

摘要：

目的：研究谷胱甘肽过氧化物酶-3（glutathione peroxidase-3，GPX-3）基因及蛋白在腮腺良、恶性多形性腺瘤中的表达，以明确GPX-3与腮腺多形性腺瘤发生、发展的相关性，为临床预测腮腺多形性腺瘤的发生及恶变提供理论依据。方法：采用荧光定量PCR 和蛋白免疫印迹法检测30例腮腺多形性腺瘤，30例腮腺多形性腺瘤的瘤旁2 cm腺体组织，以及10例恶性腮腺多形性腺瘤中的GPX-3 mRNA及蛋白的表达，对其相对表达量进行统计学分析。结果：GPX-3 mRNA和蛋白在腮腺良、恶性多形性腺瘤中的表达明显低于瘤旁腺体组织，且在腮腺恶性多形性腺瘤中的表达最低，差异有统计学意义（P<0.01）。GPX-3在腮腺多形性腺瘤中的表达与患者的年龄、性别及肿瘤大小无关，差异无统计学意义（P>0.05），且GPX-3在腮腺多形性腺瘤中的表达与肿瘤TNM分期及恶性成分比例有关，其差异有统计学意义（P<0.05）。结论：GPX-3在腮腺良性多形性腺瘤中低表达，且在腮腺恶性多形性腺瘤中的表达最低，提示GPX-3的低表达与腮腺多形性腺瘤的发生及恶变密切相关。

关键词: 谷胱甘肽过氧化物酶3, 腮腺多形性腺瘤, 荧光定量PCR, 蛋白免疫印迹

Abstract:

Objective: To study the expression of glutathione peroxidase-3(GPX-3) gene and protein in benign and malignant pleomorphic adenomas of the parotid gland to identify the occurrence and development of GPX-3 and parotid pleomorphic adenoma. The correlation provides a theoretical basis for the clinical prediction of the occurrence and malignant transformation of the parotid pleomorphic gland. Methods: Quantitative PCR and Western blotting were used to detect GPX-3 mRNA and protein in 30 cases of parotid pleomorphic adenoma, 30 cases of the tissues 2 cm around the parotid pleomorphic adenoma and 10 cases of malignant parotid pleomorphic adenoma. Expression differences of GPX-3 mRNA and protein were statistically analyzed. Results: The expression of GPX-3 mRNA and protein in benign and malignant pleomorphic adenomas of parotid gland was significantly lower than that in paragonadal glandular tissue, and the expression was the lowest in malignant pleomorphic adenoma of the parotid gland. The difference was statistically significant(P<0.01). The expression of GPX-3 in the pleomorphic adenoma of the parotid gland was not related to the age, sex and tumor size of the patients, and the difference was not statistically significant (P>0.05). The expression of GPX-3 in parotid pleomorphic adenoma was related to the tumor TNM stage and the proportion of malignant components with statistically significant(P<0.05). Conclusion: GPX-3 is lowly expressed in benign pleomorphic adenoma of the parotid gland and has the lowest expression in parotid pleomorphic adenoma, which suggests that the low expression of GPX-3 is closely related to the occurrence and malignant transformation of parotid pleomorphic adenoma.

Key words: glutathione peroxidase-3(GPX-3), parotid pleomorphic adenoma, real-time quantitative PCR, protein immunoblotting

中图分类号:

R782.7

张帆, 刘啸, 阿迪力·麦木提敏, 安尼卡尔·安尼瓦尔, 帕丽黛姆·图尔迪, 吴佩玲. 谷胱甘肽过氧化物酶-3在腮腺多形性腺瘤中的表达及意义[J]. 《口腔颌面外科杂志》, 2020, 30(1): 16-22.

ZHANG Fan, LIU Xiao, ADILI Maimutimin, ANNIKAER Anniwaer, PALIDAIMU Tuerdi, WU Peiling. Significance and Expression of Glutathione Peroxidase-3 in Pleomorphic Adenoma of Parotid Gland[J]. 《Journal of Oral and Maxillofacial Surgery》, 2020, 30(1): 16-22.

https://journal06.magtech.org.cn/Jweb_joms/CN/Y2020/V30/I1/16

图/表 10

图1 腮腺良性多形性腺瘤（A）和腮腺恶性多形性腺瘤（B）的组织病理学表现（×100）

Figure 1 Histopathological findings of benign pleomorphic adenoma of parotid gland (A) and malignant pleomorphic adenoma of parotid gland (B) （×100）

表1 荧光定量PCR引物序列

Table 1 Real-time PCR primer sequences

引物名称	序列
GPX3	F:5′-GGCTGGAGCATGAGATGGTAACTG-3′ R:5′-CGCTACACCACGCATACACTCTG-3′
β-actin	R:5′-CGCTACACCACGCATACACTCTG-3′ R:5′-GGGCCGGACTCGTCATAC-3′

表2 GPX-3 mRNA在瘤旁腺体组织及腮腺良恶性多形性腺瘤中的表达结果（例）

Table 2 Expression of GPX-3 mRNA in para-tumor glandular tissue and benign and malignant parotid pleomorphic adenoma（case）

分组	总例数	阳性表达	阴性表达
正常组	30	28	2
良性组	30	13	17
恶性组	10	3	7

图2 荧光定量PCR分析GPX-3基因在瘤旁腺体组织及腮腺良恶性多形性腺瘤中的相对表达量

Figure 2 Quantitative PCR analysis of GPX-3 gene in paraneoplastic glandular tissue， benign and malignant parotid pleomorphic adenoma

图3 腮腺良恶性多形性腺瘤中GPX-3基因相对瘤旁腺体组织的表达分布

Figure 3 Expression distribution of GPX-3 gene in benign and malignant pleomorphic adenomas of the parotid gland relative to adjacent tumor tissue

表3 GPX-3蛋白在瘤旁腺体组织、腮腺良恶性多形性腺瘤中的表达结果（例）

Table 3 Expression of GPX-3 protein in paraneoplastic glandular tissue, benign parotid pleomorphic adenoma, malignant parotid pleomorphic adenoma（case）

组别	总例数	阳性表达	阴性表达
正常组	30	28	2
良性组	30	13	17
恶性组	10	3	7

图4 蛋白免疫印迹法分析GPX-3蛋白在瘤旁腺体组织及腮腺良恶性多形性腺瘤中的表达

Figure 4 Western blot analysis of GPX-3 protein expression in para-glandular glandular tissue, benign parotid pleomorphic adenoma, malignant parotid pleomorphic adenoma

图5 GPX-3蛋白在瘤旁腺体组织及腮腺良恶性多形性腺瘤中的表达分布

Figure 5 GPX-3 protein expression and distribution in benign and malignant pleomorphic adenomas of the parotid glands and parotid glands

表4 GPX-3 mRNA与腮腺多形性腺瘤临床病理特征的关系（例）

Table 4 Relationship between GPX-3 mRNA and clinicopathological features of parotid pleomorphic adenoma（case）

临床资料	总例数	GPX-3 mRNA		阳性率/%	χ²	P
临床资料	总例数	+	-	阳性率/%	χ²	P
性别男	23	10	13	43.5	0.273	0.747
女	17	6	11	35.3
年龄/岁 <50	18	6	12	33.3	0.606	0.526
≥50	22	10	12	45.5
肿瘤大小/cm ≥2	14	6	8	42.9	0.730	1.000
<2 TNM分期 Ⅰ、Ⅱ Ⅲ、Ⅳ 侵袭性非侵袭性、微侵袭性明显侵袭性组织学分级 Ⅰ、Ⅱ Ⅲ 分化程度中低分化高分化恶性成分比例/% ≤50 >50	26 6 4 7 3 8 2 3 7 2 8	10 3 0 1 2 3 0 1 2 2 1	16 3 4 6 1 5 2 2 5 0 7	38.5 50.0 0 14.3 66.7 37.5 0 33.3 28.6 100.0 12.5	3.900 2.657 1.632 0.022 6.189	0.048 0.103 0.201 0.881 0.013

表5 GPX-3 蛋白与腮腺多形性腺瘤临床病理特征的关系（例）

Table 5 Relationship between GPX-3 protein and clinicopathological features of parotid pleomorphic adenoma（case）

临床资料	总例数	GPX-3蛋白		阳性率/%	χ²	P
临床资料	总例数	+	-	阳性率/%	χ²	P
性别男	23	8	15	34.8	0.614	0.522
女	17	8	9	47.1
年龄/岁 <50	18	7	11	38.9	0.170	1.000
≥50	22	9	13	40.9
肿瘤大小/cm ≥2	14	5	9	35.7	0.165	0.746
<2 TNM分期 Ⅰ、Ⅱ Ⅲ、Ⅳ 侵袭性非侵袭性、微侵袭性明显侵袭性组织学分级 Ⅰ、Ⅱ Ⅲ 分化程度中低分化高分化恶性成分比例/% ≤50 >50	26 6 4 7 3 8 2 3 7 2 8	11 3 0 0 3 2 1 0 3 2 1	15 3 4 7 0 6 1 3 4 0 7	42.3 50.0 0 0 100.0 25.0 50.0 0 42.9 100.0 12.5	3.900 12.217 0.447 2.657 6.189	0.048 0.008 0.504 0.103 0.013

参考文献 21

[1]	Seifert G, Sobin LH. The World Health Organization' histological classification of salivary gland tumors[J]. Cancer, 1992, 70(2): 379-385. doi: 10.1002/1097-0142(19920715)70:2<379::aid-cncr2820700202>3.0.co;2-c pmid: 1617588
[2]	丁高峰, 郭雷鸣, 陆寓非, 等. 磷酸烯醇式丙酮酸羧激酶和纤溶酶原激活物抑制剂1在腮腺多形性腺瘤中的表达及其意义[J]. 实用医学杂志, 2019, 35(03): 398-402.
[3]	Zhou C, Hu H, Zheng Z, et al. Association between GPX-3 promoter methylation and malignant tumors: a meta-analysis[J]. Pathol Res Pract, 2019, 215(7):152443. doi: 10.1016/j.prp.2019.152443 URL
[4]	Liu K, Jin M, Xiao L, et al. Distinct prognostic values of mRNA expression of glutathione peroxidases in non-small cell lung cancer[J]. Cancer Manag Res, 2018, 2018(10):2997-3005.
[5]	Agnani D, Camachovanegas O, Camacho C, et al. Decreased levels of serum glutathione peroxidase 3 are associated with papillary serous ovarian cancer and disease progression[J]. J Ovarian Res, 2011, 4(1):18. doi: 10.1186/1757-2215-4-18
[6]	张志愿. 口腔颌面外科学[M]. 7版. 北京: 人民卫生出版社, 2015: 367-368.
[7]	张敏, 贾志宇, 刘树妍, 等. 腮腺良性肿瘤浅叶部分切除术与浅叶切除术的循证医学分析[J]. 临床耳鼻咽喉头颈外科杂志, 2019, 33(9):875-882.
[8]	程晗菲, 尹林, 孟令旭, 等. MMP-9和VEGF在腮腺多形性腺瘤中的表达研究[J]. 口腔医学, 2015, 35(04): 258-261.
[9]	刘志明, 张虹, 黄丽, 等. PTEN基因表达与腮腺多形性腺瘤的发生发展的关系[J]. 中南医学科学杂志, 2016, 44(05):540-543,547.
[10]	Goetting H, Hermann KüHN. Glutathione peroxidases and redox-regulated transcription factors[J]. Biol Chem, 2006, 387(10-11):1329-1335.
[11]	Jeong W, Bae SH, Toledano MB, et al. Role of sulfiredoxin as a regulator of peroxiredoxin function and regulation of its expression[J]. Free Radic Biol Med, 2012, 53(3):447-456. doi: 10.1016/j.freeradbiomed.2012.05.020 URL
[12]	Brigeliusflohé R, Maiorino M. Glutathione peroxidases[J]. Biochim Biophys Acta, 2013, 1830(5): 3289-3303. doi: 10.1016/j.bbagen.2012.11.020 pmid: 23201771
[13]	张漫丽, 张国君, 刘静, 等. 谷胱甘肽过氧化物酶-3在恶性肿瘤发生发展中作用的研究进展[J]. 中华实验外科杂志, 2019, 36(7):1337-1340.
[14]	Zhou C, Pan R, Li B, et al. GPX-3 hypermethylation in gastric cancer and its prognostic value in patients aged over 60[J]. Future Oncol, 2019, 1 5(11): 1279-1289.
[15]	Liu Q, Bai W, Huang F, et al. Downregulation of microRNA-196a inhibits stem cell self-renewal ability and stemness in non-small-cell lung cancer through upregulating GPX-3 expression[J]. Int J Biochem Cell Biol, 2019, 115:105571. doi: 10.1016/j.biocel.2019.105571 URL
[16]	Zhao H, Li J, Li X, et al. Silencing GPX-3 expression promotes tumor metastasis in human thyroid cancer[J]. Curr Protein Pept Sci, 2015, 16(4): 316-321. doi: 10.2174/138920371604150429154840 URL
[17]	Pelosof L, Yerram S, Armstrong T, et al. GPX-3 promoter methylation predicts platinum sensitivity in colorectal cancer[J]. Epigenetics, 2017, 12(7): 540-550. doi: 10.1080/15592294.2016.1265711 URL
[18]	Chen H, Zheng Z, Kim KY, et al. Hypermethylation and downregulation of glutathione peroxidase 3 are related topathogenesis of melanoma[J]. Oncol Rep, 2016, 36(5):2737-2744. doi: 10.3892/or.2016.5071 URL
[19]	Guilleret I, Losi L, Chelbi ST, et al. DNA methylation profiling of esophageal adenocarcinoma using methylation ligation-dependentmacroarray(MLM)[J]. Biochem Biophys Res Commun, 2016, 479(2):231-237. doi: 10.1016/j.bbrc.2016.09.049 URL
[20]	Min SY, Kim HS, Jung EJ, et al. Prognostic significance of glutathione peroxidase 1 (GPX1) down-regulation andcorrelation with aberrant promoter methylation in human gastric cancer[J]. Anticancer Res, 2012, 32(8), 3169-3175.
[21]	Yi Z, Jiang L, Zhao L, et al. Glutathione peroxidase 3 (GPX-3) suppresses the growth of melanoma cells through reac tive oxygen species (ROS)-dependent stabilization of hypoxia-inducible factor 1-α and 2-α[J]. J Cell Biochem, 2019, 120(11):19124-19136. doi: 10.1002/jcb.v120.11 URL

选择文件类型/文献管理软件名称

选择包含的内容