上调Decorin对口腔鳞癌荷瘤裸鼠EGFR、C-Myc、p21表达的影响

doi:10.19439/j.sjos.2023.01.008

上海口腔医学 ›› 2023, Vol. 32 ›› Issue (1): 40-46.doi: 10.19439/j.sjos.2023.01.008

上调Decorin对口腔鳞癌荷瘤裸鼠EGFR、C-Myc、p21表达的影响

郭梦瑶^1,2, 李凯玉^1,2, 聂敏海^1,2,*, 刘旭倩^1,2,*

1.西南医科大学附属口腔医院牙周黏膜病科,四川泸州 646000;
2.西南医科大学口颌面修复重建和再生泸州市重点实验室, 西南医科大学口腔医学研究所,四川泸州 646000

收稿日期:2021-09-07 修回日期:2021-11-03 出版日期:2023-02-25 发布日期:2023-06-12
通讯作者: 聂敏海,E-mail:nieminhai@126.com;刘旭倩,E-mail：liuxuqiankokky@126.com。^*共同通信作者
作者简介:郭梦瑶（1994-）,女,硕士,医师,E-mail:2486828365@qq.com
基金资助:
四川省科学技术厅中央引导地方科技发展项目（2021ZYD0083）; 四川省科技计划项目（2022NSFC0716）; 西南医科大学应用基础重点项目（2021ZKZD010）

Effect of up-regulation of Decorin on expression of EGFR, C-Myc and p21 in nude mice with oral squamous cell carcinoma

GUO Meng-yao^1,2, LI Kai-yu^1,2, NIE Min-hai^1,2, LIU Xu-qian^1,2

1. Department of Periodontal Mucosal Diseases, the Affiliated Stomatological Hospital of Southwest Medical University. Luzhou 646000;
2. Oral & Maxillofacial Reconstruction and Regeneration of Luzhou Key Laboratory, Institute of Stomatology, Southwest Medical University. Luzhou 646000, Sichuan Province, China

Received:2021-09-07 Revised:2021-11-03 Online:2023-02-25 Published:2023-06-12

摘要/Abstract

摘要： 目的: 探讨饰胶蛋白聚糖基因（decorin,DCN）过表达对口腔鳞癌（oral squamous cell carcinoma,OSCC）裸鼠荷瘤中表皮生长因子受体（epidermal growth factor receptor,EGFR）、骨髓细胞瘤病毒癌基因（cellular-myelocytomatosis viral oncogene,C-Myc）和细胞周期蛋白依赖性激酶抑制剂（cyclin dependent kinase inhibitor,p21）表达水平的影响。方法: 通过脂质体转染法上调人口腔鳞癌细胞（HSC-3）中DCN基因的表达,使用裸鼠作为OSCC疾病模型构建载体,建立OSCC裸鼠荷瘤模型。H-E法确定各组裸鼠荷瘤组织的病理分级,免疫组织化学染色检测DCN过表达后各组裸鼠荷瘤组织中EGFR、C-Myc和p21蛋白的表达差异,RT-qPCR及Western印迹定量检测DCN过表达后各组裸鼠荷瘤组织中EGFR、C-Myc和p21在转录水平和蛋白水平上的表达差异,明确DCN过表达对OSCC裸鼠荷瘤组织中EGFR、C-Myc和p21表达的影响。采用SPSS 20.0软件包对数据进行统计学分析。结果: H-E染色结果显示,OSCC动物疾病模型构建成功。重量分析结果显示,质粒组裸鼠荷瘤组织轻于空载组和未转染组（P<0.05）。IHC结果显示,DCN、EGFR、C-Myc和p21蛋白在各组裸鼠荷瘤组织中均有表达,质粒组中DCN、EGFR和C-Myc蛋白的表达水平差异有统计学意义（P<0.05）,各组中p21蛋白的表达水平无统计学差异（P>0.05）。RT-qPCR和Western印迹结果显示,DCN、EGFR、C-Myc和p21在各组裸鼠荷瘤组织中均有不同程度表达（P<0.05）。结论: 上调DCN可抑制OSCC裸鼠荷瘤的生长。在OSCC裸鼠荷瘤组织中,DCN过表达下调EGFR及C-Myc的表达,上调p21的表达。DCN可能通过以上机制调节OSCC的发生、发展,发挥抑癌作用。

关键词: 饰胶蛋白聚糖, 口腔鳞癌, 表皮生长因子受体, 骨髓细胞瘤病毒癌基因, 细胞周期蛋白依赖性激酶抑制剂

Abstract: PURPOSE: To explore the effect of overexpression of DCN（decorin） gene on the expression of epidermal growth factor receptor (EGFR), cellular-myelocytomatosis viral oncogene (C-Myc) and cyclin dependent kinase inhibitor (p21)in tumor-bearing nude mice with oral squamous cell carcinoma（OSCC）. METHODS: The expression of DCN gene in human oral squamous cell carcinoma(HSC-3) was up-regulated by liposome transfection. Nude mice were used as the carrier of OSCC. H-E staining was used to determine the pathological grade of tumor-bearing tissues in each group. Immunohistochemistry was used to detect the expression of EGFR, C-Myc and p21 protein in tumor-bearing tissues of each group after DCN overexpression. RT-qPCR and Western blot were used to quantitatively detect the expression of EGFR, C-Myc and p21 in tumor-bearing tissues of each group after DCN overexpression, and to determine the effects of DCN overexpression on the expression of EGFR, C-Myc and p21 in tumor-bearing tissues of OSCC nude mice. SPSS 20.0 software package was used for statistical analysis. RESULTS: H-E staining showed that the animal model of OSCC was successfully constructed. The tumor-bearing tissues of nude mice in the plasmid group were significantly lighter than those in the empty vector group and non-transfected group(P<0.05). IHC results showed that DCN, EGFR, C-Myc and p21 proteins were expressed in the tumor-bearing tissues of nude mice in each group, the expression of DCN,EGFR and C-Myc proteins in the plasmid group was significantly different from the other groups(P<0.05).There was no significant differece in p21 protein expression in each group(P>0.05). RT-qPCR and Western blot results showed that DCN, EGFR, C-Myc and p21 were expressed in diffrent degrees in tumor-bearing tissues of nude mice（P<0.05）. CONCLUSIONS: DCN can inhibit the growth of tumor in OSCC nude mice. In tumor-bearing tissues of nude mice with OSCC, overexpression of DCN can down-regulate the expression of EGFR and C-Myc, and up-regulate the expression of p21.DCN may play an inhibitory role in the occurrence and development of OSCC.

Key words: Decorin, Oral squamous cell carcinoma, Epidermal growth factor receptor, Cellular-myelocytomatosis viral oncogene, Cyclin dependent kinase inhibitor

中图分类号:

R739.8

郭梦瑶, 李凯玉, 聂敏海, 刘旭倩. 上调Decorin对口腔鳞癌荷瘤裸鼠EGFR、C-Myc、p21表达的影响[J]. 上海口腔医学, 2023, 32(1): 40-46.

GUO Meng-yao, LI Kai-yu, NIE Min-hai, LIU Xu-qian. Effect of up-regulation of Decorin on expression of EGFR, C-Myc and p21 in nude mice with oral squamous cell carcinoma[J]. Shanghai Journal of Stomatology, 2023, 32(1): 40-46.

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上调Decorin对口腔鳞癌荷瘤裸鼠EGFR、C-Myc、p21表达的影响

Effect of up-regulation of Decorin on expression of EGFR, C-Myc and p21 in nude mice with oral squamous cell carcinoma

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