中国病理生理杂志2024,Vol.40Issue(1) :47-57.DOI:10.3969/j.issn.1000-4718.2024.01.006

RhoC基因沉默对口腔鳞状细胞癌迁移和侵袭的影响

Effect of RhoC silencing on migration and invasion of oral squamous cell carcinoma

杨杰 李欢 王鑫 陈正岗 王莹 李泉 华英杰 王婧 迟增鹏
中国病理生理杂志2024,Vol.40Issue(1) :47-57.DOI:10.3969/j.issn.1000-4718.2024.01.006

RhoC基因沉默对口腔鳞状细胞癌迁移和侵袭的影响

Effect of RhoC silencing on migration and invasion of oral squamous cell carcinoma

杨杰 1李欢 2王鑫 3陈正岗 4王莹 5李泉 6华英杰 1王婧 1迟增鹏7
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作者信息

  • 1. 潍坊医学院附属医院口腔科,山东 潍坊 261031
  • 2. 潍坊市人民医院口腔科,山东 潍坊 261500
  • 3. 长春中医药大学,吉林 长春 130117
  • 4. 康复大学青岛医院(青岛市市立医院),山东 青岛 266071
  • 5. 济南市第四人民医院口腔科,山东 济南 250031
  • 6. 潍坊医学院附属医院中医科,山东 潍坊 261031
  • 7. 青岛西海岸新区中心医院口腔科,山东 青岛 266000
  • 折叠

摘要

目的:分析RhoC对口腔鳞状细胞癌(OSCC)细胞迁移和侵袭能力的影响及其潜在机制.方法:利用UALCAN和K-M plotter在线数据库预测分析RhoC在OSCC中的表达及其与病理分期分生存率的关系,并结合IHC实验检测OSCC组织中RhoC的表达水平.按照人RhoC基因构建两条小干扰RNA(siRNA)并将细胞进行实验分组.RT-qPCR检测转染后各组细胞RhoC的mRNA表达水平,Western blot法分析实验分组细胞中RhoC、FAK、MAPK、p-FAK、p-MAPK、基质金属蛋白酶2(MMP-2)和MMP-9蛋白表达;此外,比较各实验分组细胞的穿膜细胞数和划痕愈合面积分析细胞的侵袭和迁移活动.最终通过构建裸鼠肺转移模型对上述实验进行验证.结果:RhoC在OSCC中高表达,并和病理分期和病理分级等关联性大.RhoC的mRNA和蛋白相对表达量在OSCC中增高(P<0.01);RT-qPCR及Western blot结果显示,敲减RhoC表达后,RhoC的mRNA和蛋白表达水平显著下降(P<0.01),Western blot结果显示p-FAK、p-MAPK、MMP-2和MMP-9蛋白水平显著下降(P<0.05),FAK和MAPK蛋白表达无明显变化(P>0.05);细胞迁移和侵袭能力显著下降(P<0.01);实验组在动物活体荧光成像系统下发出的荧光强度显著低于对照组,且HE染色结果显示实验组裸鼠肺结节数量明显减少(P<0.05).结论:RhoC基因可影响OSCC细胞的迁移和侵袭能力,其潜在机制可能是通过活化FAK和MAPK信号分子参与OSCC细胞的迁移和侵袭过程.

Abstract

AIM:To explore the expression of RhoC in oral squamous cell carcinoma(OSCC)and its effects on the malignant biological behavior of OSCC cells.METHODS:The UALCAN and K-M plotter databases,alongside tis-sue sample analyses,facilitated understanding RhoC expression in cancer and its links to clinicopathological traits.Two small interfering RNAs(RhoC-siRNA)were constructed according to the RhoC gene sequence.The mRNA and protein ex-pression levels of RhoC in OSCC cells were determined.The protein levels of FAK,p-FAK,MAPK,p-MAPK,matrix me-talloproteinase-2(MMP-2)and MMP-9 were also examined by Western blot.Furthermore,the invasion and migration of OSCC cells were analyzed by Transwell assay and scratch test.Finally,the pulmonary metastasis model of nude mice was established.RESULTS:The results of the databases showed that RhoC was highly expressed in OSCC tissues,which was closely related to pathological stage,pathological grade and lymph node metastasis,but not significantly related to the sur-vival rate of patients.Furthermore,compared with paracancer tissues,the mRNA and protein expression levels of RhoC were increased in OSCC tissues(P<0.01).Silencing of RhoC prominently reduced the migration and invasion of OSCC cells as well as the protein levels of p-FAK,p-MAPK,MMP2 and MMP9(P<0.05).The protein levels of MAPK and FAK were unchanged(P>0.05).The fluorescence intensity of the experimental group was significantly lower than that of the control group,and the results of HE staining showed that the number of lung nodules in the experimental group was sig-nificantly reduced(P<0.05).CONCLUSION:RhoC can effectively influence the migration and invasion of OSCC cells,and its potential mechanism may be related to FAK/MAPK/MMPs signaling pathway.

关键词

口腔鳞状细胞癌/RhoC基因/小干扰RNA/细胞迁移/细胞侵袭

Key words

oral squamous cell carcinoma/RhoC gene/small interfering RNA/cell migration/cell invasion

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基金项目

国家自然科学基金资助项目(81372908)

山东省中医药科技发展计划项目(2019-0440)

济南市卫计委科技计划(2017-2-02)

出版年

2024
中国病理生理杂志
中国病理生理学会

中国病理生理杂志

CSTPCD北大核心
影响因子:1.065
ISSN:1000-4718
被引量1
参考文献量2
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