华西口腔医学杂志 ›› 2021, Vol. 39 ›› Issue (5): 510-517.doi: 10.7518/hxkq.2021.05.003

• 基础研究 • 上一篇    下一篇

RhoE在舌鳞状细胞癌中的表达及其对细胞迁移和侵袭的影响

赵开1(), 袁文红2, 李文健3, 迟增鹏4, 王少如3, 陈正岗2()   

  1. 1.青岛大学口腔医学院,青岛 266003
    2.青岛大学附属青岛市市立医院口腔医学中心,青岛 266071
    3.大连医科大学口腔医学院,大连 116044
    4.潍坊医学院口腔医学院,潍坊 261021
  • 收稿日期:2020-12-10 修回日期:2021-06-28 出版日期:2021-10-01 发布日期:2021-10-11
  • 通讯作者: 陈正岗 E-mail:1142083797@qq.com;chenzhg1973@163.com
  • 作者简介:赵开,硕士,E-mail:1142083797@qq.com
  • 基金资助:
    国家自然科学基金面上项目(81372908)

Effect of RhoE expression on the migration and invasion of tongue squamous cell carcinoma

Zhao Kai1(), Yuan Wenhong2, Li Wenjian3, Chi Zengpeng4, Wang Shaoru3, Chen Zhenggang2()   

  1. 1.School of Stomatology, Qingdao University, Qingdao 266003, China
    2.Dept. of Stomatology, Qingdao Municipal Hospital, Qingdao University, Qingdao 266071, China
    3.School of Stomatology, Dalian Medical University, Dalian 116044, China
    4.College of Stomatology, Weifang Medical University, Weifang 261021, China
  • Received:2020-12-10 Revised:2021-06-28 Online:2021-10-01 Published:2021-10-11
  • Contact: Chen Zhenggang E-mail:1142083797@qq.com;chenzhg1973@163.com
  • Supported by:
    The National Natural Science Foundation of China(81372908);Correspondence: Chen Zhenggang, E-mail: chenzhg1973@163.com

摘要: 目的

初步探讨RhoE在舌鳞状细胞癌(TSCC)中的表达水平,及其对TSCC细胞迁移和侵袭的影响及相关机制。

方法

选取在青岛大学附属青岛市市立医院口腔医学中心颌面外科2017—2019年手术切除的TSCC原发灶48例,采用免疫组织化学法检测标本(TSCC组织、癌旁组织)中RhoE的表达水平,并提取标本的RhoE mRNA和蛋白进一步检测RhoE的表达水平。选取TSCC SCC-4和CAL-27细胞系进行体外实验,采用瞬时转染法过表达RhoE,并应用实时荧光定量PCR(qRT-PCR)及蛋白质免疫印记法检测过表达效率;细胞划痕实验及Transwell细胞侵袭实验分别检测TSCC细胞的迁移和侵袭能力;蛋白质免疫印记法检测相关蛋白Rho关联含卷曲螺旋结合蛋白激酶1(ROCK1)、基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)的表达水平;采用GraphPad Prism 8.2.1软件对实验数据进行统计学分析。

结果

TSCC组织中RhoE的表达水平显著低于癌旁组织(P<0.05);在体外实验中,与空白对照组和阴性对照组相比,过表达实验组TSCC细胞的迁移和侵袭能力显著降低(P<0.05),过表达实验组ROCK1、MMP-2及MMP-9表达显著降低(P<0.05)。

结论

RhoE在TSCC组织中低表达,上调TSCC细胞中RhoE表达可显著抑制其迁移和侵袭能力,提示RhoE可能在调控TSCC转移、侵袭等恶性生物学行为过程中发挥作用,为TSCC的基因治疗提供新的靶点。

关键词: RhoE, 舌鳞状细胞癌, 细胞迁移, 细胞侵袭

Abstract: Objective

This study aims to investigate the effect of RhoE expression on the migration and invasion of tongue squamous cell carcinoma (TSCC).

Methods

Forty-eight TSCC cases were selected from the Maxillofacial Surgery Center of Qingdao Municipal Hospital from 2017 to 2019. The expression of RhoE in the specimens (TSCC and adjacent tissues) was detected by immunohistochemistry, and RhoE mRNA and protein were extracted to further detect the expression of RhoE. SCC-4 and CAL-27 cells were selected for in vitro experiments. Transient transfection was used to overexpress RhoE. Real-time fluorescence quantitative PCR (qRT-PCR) and Western blot analyses were conducted to detect the overexpression efficiency. Scratch test and Transwell cell invasion tests were used to detect the migration and invasion ability of TSCC, respectively. The expression levels of Rho-associated coiled-coil-containing protein kinase 1 (ROCK1), matrix metalloproteinase-2 (MMP-2), and matrix metalloproteinase-9 (MMP-9) were detected by Western blot. Experimental data were analyzed by Graphpad prism 8.2.1 software.

Results

The expression level of RhoE in TSCC was significantly lower than that in adjacent tissues (P<0.05). The migration and invasion abilities of TSCC were significantly lower than those in the control group (P<0.05). The Western blot showed significantly lower expression levels of ROCK1, MMP-2, and MMP-9 in the experimental group than in the control group (P<0.05).

Conclusion

RhoE expression is low in TSCC. Over expression RhoE in TSCC can significantly decrease its migration and invasion abilities. Hence, RhoE may play an important role in regulating the metastasis and invasion of TSCC and provide a new target for gene therapy.

Key words: RhoE, tongue squamous cell carcinoma, migration, invasion

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