国际口腔医学杂志

国际口腔医学杂志 ›› 2026, Vol. 53 ›› Issue (4): 526-536.doi: 10.7518/gjkq.2026228

• 论著 • 上一篇    

5-羟色胺受体2C通过Gαq/11-Yes相关蛋白信号通路调控口腔鳞状细胞癌的作用研究

张真语(),黄梅,崔浩,孙思露,罗小波,江潞,江宇辰()   

  1. 口腔疾病防治全国重点实验室 国家口腔医学中心 口腔疾病国家临床医学研究中心中国医学科学院口腔黏膜癌变与防治创新单元 四川大学华西口腔医院口腔黏膜病科 成都 610041
  • 收稿日期:2025-02-18 修回日期:2025-12-23 出版日期:2026-07-01 发布日期:2026-06-25
  • 通讯作者: 江宇辰
  • 作者简介:张真语，硕士，Email：zhenyuzhang7@outlook.com
  • 基金资助:
    国家自然科学基金青年项目(82002888)

Regulation of the progression of oral squamous cell carcinoma by 5-hydroxytryptamine receptor 2C via the Gαq/11-Yes-associated protein signaling pathway

Zhenyu Zhang(),Mei Huang,Hao Cui,Silu Sun,Xiaobo Luo,Lu Jiang,Yuchen Jiang()   

  1. State Key Laboratory of Oral Diseases & National Center for Stomatology & National Clinical Research Center for Oral Diseases & Chinese Academy of Medical Sciences Research Unit of Oral Carcinogenesis and Management & Dept. of Oral Medicine, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China
  • Received:2025-02-18 Revised:2025-12-23 Online:2026-07-01 Published:2026-06-25
  • Contact: Yuchen Jiang
  • Supported by:
    National Natural Science Foundation of China(82002888)

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摘要：

目的 探究5-羟色胺受体2C（HTR2C）通过Gαq/11-Yes相关蛋白（YAP）信号通路对口腔鳞状细胞癌（OSCC）进展的调控作用。 方法 利用TCGA数据库分析HTR2C在OSCC与正常口腔组织中的表达情况及其与患者总生存率的关系。通过免疫组织化学染色在OSCC肿瘤组织芯片中检测HTR2C表达水平，并分析其与临床病理特征的相关性。构建HTR2C敲低的HN12和UM1细胞系，通过细胞计数试剂盒（CCK）-8、克隆形成、划痕和Transwell实验评估HTR2C敲低或其抑制剂Puerarin对OSCC细胞增殖、迁移和侵袭的影响。采用蛋白质印迹（Western blot）、实时荧光定量聚合酶链式反应和免疫荧光检测HTR2C-Gαq/11-YAP信号通路相关蛋白的表达及激活状态。利用裸鼠OSCC皮下移植瘤模型验证HTR2C对OSCC生长的调控作用。 结果 HTR2C在OSCC组织中显著高表达，其高表达与患者不良预后显著相关（P<0.05）。体外实验表明，敲低HTR2C或使用Puerarin显著抑制OSCC细胞的增殖、克隆形成、迁移和侵袭能力（P<0.01），且Puerarin呈剂量依赖性抑制作用。机制研究发现，HTR2C敲低后通过减少黏着斑激酶和丝切蛋白的磷酸化，抑制YAP在Tyr357位点的磷酸化，促进YAP Ser127位点磷酸化，从而抑制YAP的转录活性，降低其下游靶基因CYR61、CTGF和AURKA的表达（P<0.05）。体内实验证实，通过Puerarin抑制HTR2C的表达，可显著抑制OSCC移植瘤的生长（P<0.000 1），这一作用是通过调控YAP信号通路实现的。 结论 OSCC中HTR2C的高表达与患者不良预后显著相关，HTR2C可能经激活Gαq/11-YAP信号通路，调控OSCC细胞的增殖、迁移、侵袭以及体内成瘤能力。本研究提示，HTR2C可能是OSCC治疗的潜在靶点，但仍需进一步的临床研究加以验证。

关键词: 口腔鳞状细胞癌, 5-羟色胺受体2C, Gαq/11-Yes 相关蛋白信号通路, Yes 相关蛋白, Puerarin, 肿瘤进展

Abstract:

Objective　To investigate the regulatory effect of 5-hydroxytryptamine receptor 2C (HTR2C) on oral squamous cell carcinoma (OSCC) through the Gαq/11-Yes-associated protein (YAP) signaling pathway.Me-thods　The TCGA database was used to analyze the expression of HTR2C in OSCC and normal oral tissues and its relationship with the overall survival rate of patients. The expression level of HTR2C was detected in OSCC tumor microarrays through immunohistochemistry, and the correlation between HTR2C and clinicopathological features was analyzed. HTR2C knockdown HN12 and UM1 cell lines were constructed, and the effects of HTR2C knockdown or its inhi-bitor (Puerarin) on the proliferation, migration, and invasion of OSCC cells were evaluated through cell counting kit-8, colony formation, wound healing, and Transwell assays. Western blot analysis, quantitative real-time polymerase chain reaction, and immunofluorescence were used to detect the expression and activation of the HTR2C-Gαq/11-YAP signaling pathway. The effect of HTR2C on the growth of OSCC was verified by using a subcutaneous xenograft model of OSCC in nude mice. Results　HTR2C was significantly overexpressed in OSCC tissues, and its high expression was significantly associated with the poor prognosis of patients (P<0.05). In vitro experiments demonstrated that the knockdown of HTR2C or use of Puerarin significantly inhibited the proliferation and clonal formation, migration, and invasion abilities of OSCC cells (P<0.01), and the inhibitory effect of puerarin was dose-dependent. Mechanism studies showed that HTR2C knockdown inhibited YAP phosphorylation at Tyr357 and promoted YAP phosphorylation at Ser127 by reducing the phosphorylation of focal adhesion kinase and Cofilin, thereby inhibiting YAP transcriptional activity and decreasing the expression of its downstream target genes CYR61, CTGF, and AURKA (P<0.05). In vivo experiments confirmed that inhibiting the expression of HTR2C by using Puerarin could significantly inhibit the growth of OSCC xenograft tumors (P<0.000 1). This effect was achieved by regulating the YAP signaling pathway. Conclusion　The high expression of HTR2C in OSCC was significantly associated with poor prognosis. HTR2C may regulate the proliferation, migration, invasion, and tumorigenesis of OSCC cells by activating the Gαq/11-YAP signaling pathway. This study suggests that HTR2C may be a potential therapeutic target for OSCC. However, further clinical research is needed to validate its findings.

Key words: oral squamous cell carcinoma, 5-hydroxytryptamine receptor 2C, Gαq/11-Yes-associated protein signaling pathway, Yes-associated protein, Puerarin, tumor progression

中图分类号: 

  • R739.8

图 1

OSCC及对照组织中HTR2C与YAP的IHC染色结果代表图A：IHC染色结果中HTR2C表达的4种情况代表图； B：IHC染色结果中YAP表达的4种情况代表图。"

图2

OSCC 组织中 HTR2C 的表达及其与预后的相关性A：通过TCGA数据库评估OSCC 组织和正常对照组织中HTR2C 表达差异；B：使用TCGA数据库数据绘制的Kaplan-Meier生存曲线；C：组织芯片中HTR2C在OSCC组织和正常对照组织中的表达差异；D：组织芯片中 HTR2C 表达情况与 OSCC 病理等级的相关性；E：WB检测OSCC细胞系HTR2C的表达水平（n=3）；与对照组相比，*为P<0.05，***为P<0.001。"

图3

体外敲低HTR2C可显著抑制OSCC细胞增殖和迁移侵袭A：WB检测HN12和UM1细胞中HTR2C表达情况（n=3）；B：HN12和 UM1细胞的CCK-8实验（n=3）；C：HN12和UM1细胞克隆形成实验结果和统计分析（n=3）；D：HN12和UM1细胞划痕试验结果和统计分析（n=3）；E：HN12和UM1细胞Transwell 侵袭实验结果和统计分析（n=3）。与对照组相比，**为P<0.01，***为P<0.001，****为P<0.000 1。"

图4

HTR2C可通过激活Gαq/11-YAP信号通路促进OSCC细胞的恶性进展A：组织芯片中YAP在OSCC组织和正常对照组织中的表达差异；B：组织芯片中YAP表达情况与OSCC病理等级的相关性；C：组织芯片中YAP表达情况与HTR2C表达情况的相关性；D：WB检测 HN12和UM1细胞中 YAP 信号通路蛋白水平（n=3）；E：qRT-PCR 检测 HN12和UM1细胞中YAP信号通路 mRNA 表达水平（n=3）；F：细胞免疫荧光实验检测HN12和UM1细胞中YAP蛋白的核质分布情况及统计分析（n=3）。与对照组相比，*为P<0.05；**为P<0.01；***为P<0.001；****为P<0.000 1。"

图 5

HTR2C抑制剂显著抑制OSCC细胞的增殖、迁移和侵袭A：HTR2C抑制剂抑制HN12和UM1细胞生长的IC50实验；B：WB检测HTR2C抑制剂处理对细胞蛋白表达的影响（n=3）；C：HTR2C抑制剂处理HN12和 UM1细胞克隆形成实验结果及统计分析（n=3）；D：HTR2C抑制剂处理HN12和UM1细胞划痕试验结果及统计分析（n=3）；E：HTR2C抑制剂处理HN12和UM1细胞Transwell侵袭实验结果及统计分析（n=3）。与对照组相比，**为P<0.01；***为P<0.001；****为P<0.000 1。"

图6

HTR2C抑制剂在体内有效抑制OSCC移植瘤的生长A：动物实验模式图；B：对照组与给药组的肿瘤白光照片；C：对照组与给药组的肿瘤体积随治疗时间的变化曲线（n=6）；D：对照组与给药组的小鼠体重随治疗时间的变化曲线（n=6）；E：对照组与给药组的肿瘤中YAP和Ki-67的IHC染色结果及统计分析（n=3，细胞数1 000~1 500个）；F：对照组与给药组的心、肝、脾、肺、肾的HE染色结果。与对照组相比，****为P<0.000 1，ns为差异无统计学意义。"

图7

OSCC中HTR2C调控Gαq/11-YAP信号通路的模式图"

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