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.

参考文献

[1] 张志愿. 口腔颌面外科学[M]. 北京: 人民卫生出版社, 2012: 324-329.
[2] 于世凤. 口腔组织病理学[M]. 北京: 人民卫生出版社, 2012: 380-383.
[3] Järvinen TA, Prince S.Decorin: a growth factor antagonist for tumor growth inhibition[J]. Biomed Res Int, 2015, 2015: 654765.
[4] Gao Y, Ma HY, Xu QY, et al.Mechanism of decorin protein inhibiting invasion and metastasis of non-small cell lung cancer[J]. Eur Rev Med Pharmacol Sci, 2019, 23(4): 1520-1527.
[5] 吴凡, 王千秋. 核心蛋白聚糖生物学功能研究进展[J]. 医学综述, 2017, 23(2): 249-252, 256.
[6] Kani K, Warren CM, Kaddis CS, et al.Oligomers of ERBB3 have two distinct interfaces that differ in their sensitivity to disruption by heregulin[J]. J Biol Chem, 2005, 280(9): 8238-8247.
[7] 李王丽, 佟晓, 陈斌. 宫颈鳞状细胞癌中MUC4、EGFR和p-AKT的表达及意义[J]. 实用癌症杂志, 2019, 34(7): 1068-1071.
[8] 崔忠, 赵清涛, 孙建斌, 等. 肝细胞癌患者未转染组织中survivin、PTEN、EGFR表达水平及临床意义[J]. 空军医学杂志, 2017, 33(1): 30-32.
[9] 李俏丽, 李超, 王薇, 等. 口腔、口咽鳞状细胞癌中HPV感染与EGFR之间的相关性研究[J]. 肿瘤预防与治疗, 2018, 31(2): 78-83.
[10] Zhang B, Zhang Y, Jiang X, et al.JMJD8 promotes malignant progression of lung cancer by maintaining EGFR stability and EGFR/PI3K/AKT pathway activation[J]. J Cancer, 2021, 12(4): 976-987.
[11] Jang JW, Song Y, Kim SH, et al.CD133 confers cancer stem-like cell properties by stabilizing EGFR-AKT signaling in hepatocellular carcinoma[J]. Cancer Lett, 2017,389: 1-10.
[12] Kumari R, Chouhan S, Singh S, et al.Constitutively activated ERK sensitizes cancer cells to doxorubicin: involvement of p53-EGFR-ERK pathway[J]. J Biosci, 2017, 42(1): 31-41.
[13] 汪峰, 胡玉崇, 贺凤, 等. EGFR与PD-1、PD-L1在子宫内膜癌中的表达研究[J]. 中国实验诊断学, 2020, 24(10): 1586-1589.
[14] 向青林, 张伟. HER2、EGFR、VEGFR在胃印戒细胞癌中的表达及其临床意义[J]. 医学信息, 2020, 33(6): 87-89.
[15] Pan W, Wang W, Huang J, et al.The prognostic role of c-MYC amplification in schistosomiasis-associated colorectal cancer[J]. Jpn J Clin Oncol, 2020, 50(4): 446-455.
[16] 乌守恒, 曾晓峰, 王萍, 等. 宫颈癌组织中c-myc、bcat1的表达及其临床意义[J]. 四川大学学报 (医学版), 2018, 49(5): 42-47.
[17] Li XX, Shi L, Zhou XJ, et al.The role of C-Myc-RBM38 loop in the growth suppression in breast cancer[J]. J Exp Clin Cancer Res, 2017, 36(1): 49-60.
[18] Giurisato E, Lonardi S, Telfer B, et al.Extracellular-regulated protein kinase 5-mediated control of p21 expression promotes macrophage proliferation associated with tumor growth and metastasis[J]. Cancer Res, 2020, 80(16): 3319-3330.
[19] Xiao BD, Zhao YJ, Jia XY, et al.Multifaceted p21 in carcinogenesis, stemness of tumor and tumor therapy[J]. World J Stem Cells, 2020, 12(6): 481-487.
[20] Jin Q, Lin C, Zhu X, et al.125I seeds irradiation inhibits tumor growth and induces apoptosis by Ki-67, P21, survivin, livin and caspase-9 expression in lung carcinoma xenografts[J]. Radiat Oncol, 2020, 15(1): 238-248.
[21] Balabanov D, Zhao L, Zhu Z, et al.IL-29 exhibits anti-tumor effect on pan-48 pancreatic cancer cells by up-regulation of p21 and Bax[J]. Anticancer Res, 2019, 39(7): 3493-3498.
[22] 薛云. SP1、KLF4和P21在卵巢上皮性癌的表达及意义 [D]. 青岛: 青岛大学, 2020: 8-10.
[23] Xiao Y, Deng WW, Yang LL, et al.Overexpression of p21-activated kinase 2 is correlated with high-grade oral squamous cell carcinomas[J]. Future Oncol, 2018, 14(11): 1091-1100.
[24] Bissinger O, Kolk A, Drecoll E, et al.EGFR and cortactin: markers for potential double target therapy in oral squamous cell carcinoma[J]. Exp Ther Med, 2017, 14(5): 4620-4626.
[25] Chuerduangphui J, Ekalaksananan T, Chaiyarit P, et al.Effects of arecoline on proliferation of oral squamous cell carcinoma cells by dysregulating c-Myc and miR-22, directly targeting oncostatin M[J]. PLoS One, 2018, 13(1): e0192009.
[26] 赖永先, 聂敏海, 陈潇, 等. 饰胶蛋白聚糖与口腔鳞状细胞癌发生发展的相关性研究[J]. 口腔医学研究, 2020, 36(5): 437-442.
[27] 张玉成, 王雅丽, 杜珍武, 等. 重组核心蛋白聚糖转染对HepG2细胞周期、凋亡以及P21表达的影响[J]. 中国老年学杂志, 2008, 18(3): 241-243.
[28] Seidler DG, Goldoni S, Agnew C, et al.Decorin protein core inhibits in vivo cancer growth and metabolism by hindering epidermal growth factor receptor function and triggering apoptosis via caspase-3 activation[J]. J Biol Chem, 2006, 281(36): 26408-26418.
[29] 聂鼎睿. miR--204靶向调控HGF/c—MET通路在急性髓系白血病中的作用和机制的研究 [D]. 郑州: 郑州大学, 2020: 4-8.
[30] Fan Y, Jia X, Xie T, et al.Radiosensitizing effects of c-myc gene knockdown-induced G2/M phase arrest by intrinsic stimuli via the mitochondrial signaling pathway[J]. Oncol Rep, 2020, 44(6): 2669-2677.
[31] Zhang Y, Xia M, Jin K, et al.Function of the c-Met receptor tyrosine kinase in carcinogenesis and associated therapeutic opportunities[J]. Mol Cancer, 2018, 17(1): 45-59.
[32] Georgakilas AG, Martin OA, Bonner WM. p21: a two-faced genome guardian[J]. Trends Mol Med, 2017, 23(4): 310-319.
[33] Chen Y, Lin C, Liu Y, et al.HMGB1 promotes HCC progression partly by downregulating p21 via ERK/c-Myc pathway and upregulating MMP-2[J]. Tumour Biol, 2016, 37(4): 4399-4408.

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

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	董淋升, 唐进, 程维, 冉婧璇, 刘兆博, 李雅冬. IL-6和β-catenin在口腔鳞癌中的表达及预后价值[J]. 上海口腔医学, 2023, 32(3): 280-286.
[2]	胡小媛, 王宁, 张晓野, 杨茜, 高菲, 王成勤, 尚伟, 项锋钢, 冯元勇. Chemerin对口腔鳞癌中性粒细胞浸润的影响及机制探讨[J]. 上海口腔医学, 2023, 32(2): 158-165.
[3]	文旭涛, 马振, 张翀, 陶识丞, 陈兴金, 麦华明. 不同培养条件对CD133⁺原代口腔鳞癌细胞干性维持的影响[J]. 上海口腔医学, 2022, 31(4): 343-348.
[4]	秦志明, 揭伟萍, 池彦廷, 李宏锋, 李斌斌. 基于局部可变阈值的无监督定量Ki-67细胞增殖指数模型的初步建立[J]. 上海口腔医学, 2022, 31(3): 248-254.
[5]	买尔哈巴·买合木提, 拜婕, 如斯坦木·依米提, 姚志涛. 50例口腔鳞癌患者治疗过程中营养状况改变及影响因素分析[J]. 上海口腔医学, 2022, 31(2): 205-210.
[6]	谈亦然, 赵铜超, 琚梧桐, 钟来平. 长春新碱对口腔鳞癌中Stathmin调控作用的实验研究[J]. 上海口腔医学, 2022, 31(1): 1-5.
[7]	谢非, 秦星, 童桐, 蒋英英, 张建军. IGF2BP1在口腔鳞癌细胞耐药中的作用及机制探讨[J]. 上海口腔医学, 2021, 30(5): 456-461.
[8]	李蕾, 李晓东, 庄乾伟, 葛良玉, 李志萍, 顾倩平, 白玉婷, 孟箭. 超声热疗联合TPF化疗治疗19例老年晚期口腔鳞癌的疗效分析[J]. 上海口腔医学, 2021, 30(5): 543-547.
[9]	邵小钧, 朱乔, 刘有, 韩小东, 林小臻, 席庆. 咀嚼槟榔与口腔鳞癌的相关临床病理学因素分析[J]. 上海口腔医学, 2021, 30(3): 268-272.
[10]	薛佳艺, 朱晓燕, 危常磊, 孙培, 张慧, 邓婧. 25例口腔鳞癌患者唾液代谢组学分析[J]. 上海口腔医学, 2021, 30(2): 201-205.
[11]	王树斌, 秦帅华, 易雅群, 李新明. 216例口腔鳞癌患者术后心理困扰影响因素及放疗前、后生存质量评价[J]. 上海口腔医学, 2021, 30(1): 71-76.
[12]	傅宗云, 陶峰. MMP-14、ING4和HIF-1α在60例口腔鳞癌中的表达及临床意义[J]. 上海口腔医学, 2020, 29(6): 642-646.
[13]	王学梅, 蒋勇. 双硫仑对口腔鳞癌细胞上皮-间充质转化的影响及机制探讨[J]. 上海口腔医学, 2020, 29(2): 138-145.
[14]	周璐, 龚霞, 熊屏. 载药脂质体-微泡复合物超声导向治疗头颈鳞癌的体外实验研究[J]. 上海口腔医学, 2020, 29(2): 168-173.
[15]	刘族志, 赵永兴, 林建能, 周国平. 双氢青蒿素对人口腔鳞癌细胞KBV200多药耐药的影响[J]. 上海口腔医学, 2019, 28(6): 586-590.