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学科主题: 口腔医学
题名:
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
作者: 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
发表日期: 2015-07-01
DOI: 10.1111/odi.12323
卷: 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
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
项目资助者: National Nature Science Foundation of China
WOS记录号: WOS:000355741700009
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/63328
Appears in Collections:北京大学口腔医学院_期刊论文

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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

Recommended Citation:
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.
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