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The CRISPR/Cas system inhibited the pro-oncogenic effects of alternatively spliced fibronectin extra domain A via editing the genome in salivary adenoid cystic carcinoma cells
Wang, H-C1; Yang, Y.1; Xu, S-Y2; Peng, J.1; Jiang, J-H3; Li, C-Y1
关键词salivary adenoid cystic carcinoma extra domain A cellular motility CRISPR Cas pro-oncogenic splicing knockout
刊名ORAL DISEASES
2015-07-01
DOI10.1111/odi.12323
21期:5页:608-618
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Dentistry, Oral Surgery & Medicine
资助者National Nature Science Foundation of China ; National Nature Science Foundation of China
研究领域[WOS]Dentistry, Oral Surgery & Medicine
关键词[WOS]EPITHELIAL-MESENCHYMAL TRANSITION ; PROSTATE-CANCER ; EDA SEGMENT ; EXPRESSION ; GROWTH ; CAS9 ; ADENOCARCINOMA ; ENDONUCLEASE ; SPECIFICITY ; METASTASIS
英文摘要

ObjectivesTo identify the association of fibronectin (FN) extra domain A (EDA) with the progression of salivary adenoid cystic carcinoma (SACC). Accordingly, the exclusion of EDA exon through the CRISPR/Cas9 system was investigated as the rescue for such pro-oncogenic splicing.

Materials and methodsSACC-83 cells were transiently transfected with plasmids containing recombinant EDA, and the cellular growth and motility were then accessed in vitro. Epithelial-mesenchymal transition (EMT) was investigated with immunohistochemistry, Western blot, and real-time PCR analysis. SACC tissues from 81 patients were used to access the associations between EDA+FN and clinical-pathological parameters. CRISPR/Cas9 plasmids containing sgRNA were designed and co-transfected into SACC-83 cells; the effects of EDA knockout on cellular growth and motility were then accessed.

ResultsThe recombinant EDA exhibited little effect on the proliferation of SACC cells, but significantly promoted the migration and invasion of the cells (P<0.05), accompanied with upregulated EMT (P<0.05); consistently, the expression of EDA+FN was positively associated with the metastasis, nerve invasion and recurrence of SACC (P<0.05). Furthermore, the EDA knockout from the FN gene in most SACC cells resulted in a decrease in cell motility and invasion, as well as prolonged population doubling time, compared with untreated SACC-83 cells (P<0.05).

ConclusionThe EDA domain significantly promoted the motility of SACC cells, and positively associated with the tumor progression in patients with SACC. Thus, it is a potential risk factor and also a therapeutic target for SACC. The CRISPR/Cas9 system may control a pro-oncogenic splicing process through the exclusion of EDA exon from the FN gene, leading to inhibition of motility, invasion and proliferation of cancer cells.

语种英语
所属项目编号81072214 ; 81171006
资助者National Nature Science Foundation of China ; National Nature Science Foundation of China
WOS记录号WOS:000355741700009
引用统计
被引频次:5[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/63328
专题北京大学口腔医学院
作者单位1.Peking Univ, Sch & Hosp Stomatol, Cent Lab, Beijing 100081, Peoples R China
2.Shandong Univ, Sch Stomatol, Dept Oral Implanting, Jinan 250100, Peoples R China
3.Peking Univ, Sch & Hosp Stomatol, Dept Orthodont, Beijing 100081, Peoples R China
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GB/T 7714
Wang, H-C,Yang, Y.,Xu, S-Y,et al. The CRISPR/Cas system inhibited the pro-oncogenic effects of alternatively spliced fibronectin extra domain A via editing the genome in salivary adenoid cystic carcinoma cells[J]. ORAL DISEASES,2015,21(5):608-618.
APA Wang, H-C,Yang, Y.,Xu, S-Y,Peng, J.,Jiang, J-H,&Li, C-Y.(2015).The CRISPR/Cas system inhibited the pro-oncogenic effects of alternatively spliced fibronectin extra domain A via editing the genome in salivary adenoid cystic carcinoma cells.ORAL DISEASES,21(5),608-618.
MLA Wang, H-C,et al."The CRISPR/Cas system inhibited the pro-oncogenic effects of alternatively spliced fibronectin extra domain A via editing the genome in salivary adenoid cystic carcinoma cells".ORAL DISEASES 21.5(2015):608-618.
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