当前位置: X-MOL 学术Front. Mol. Biosci. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
CRISPR-Cas13-Mediated Knockdown of lncRNA-GACAT3 Inhibited Cell Proliferation and Motility, and Induced Apoptosis by Increasing p21, Bax, and E-Cadherin Expression in Bladder Cancer
Frontiers in Molecular Biosciences ( IF 5 ) Pub Date : 2020-11-27 , DOI: 10.3389/fmolb.2020.627774
Zhongfu Zhang , Jieqing Chen , Zhongshuang Zhu , Zhongqing Zhu , Xinhui Liao , Jianting Wu , Jianli Cheng , Xintao Zhang , Hongbing Mei , Guosheng Yang

The current study is to investigate the expression pattern and biological function of long non-coding RNA Focally gastric cancer-associated transcript3 (GACAT3) in bladder cancer. Real-time quantitative qPCR was used to detect the expression level of GACAT-3 in tumor tissues and paired normal tissues. Human bladder cancer T24 and 5637 cell lines were transiently transfected with specific CRISPR-Cas13 or negative control CRISPR-Cas13. Cell migration, proliferation, and apoptosis were measured by using wound healing assay CCK-8 assay and Caspase-3 ELISA assay, respectively. The expression changes of p21, Bax, and E-cadherin after knockdown of GACAT3 were detected by using Western blot. The results demonstrated that GACAT3 was up-regulated in bladder cancer tissues than that in the paired normal tissues. Inhibition of cell proliferation, increased apoptosis, and decreased motility were observed in T24 and 5637 cell lines transfected by CRISPR-Cas13 targeting GACAT3. Downregulation of GACAT3 increased p21, Bax, and E-cadherin expression and silencing these genes could eliminate the phenotypic changes induced by knockdown of GACAT3. A ceRNA mechanism for GACAT3 was also revealed. By using CRISPR-Cas13 biotechnology, we suggested that GACAT3 may be a novel target for diagnosis and treatment of bladder cancer.



中文翻译:

CRISPR-Cas13介导的lncRNA-GACAT3抑制基因抑制膀胱癌细胞增殖和运动性,并通过增加p21,Bax和E-钙黏着蛋白的表达诱导细胞凋亡。

目前的研究是调查长非编码RNA胃癌相关转录本3(GACAT3)在膀胱癌中的表达模式和生物学功能。实时定量qPCR用于检测GACAT-3在肿瘤组织和配对的正常组织中的表达水平。用特异性CRISPR-Cas13或阴性对照CRISPR-Cas13瞬时转染人膀胱癌T24和5637细胞系。通过使用伤口愈合测定CCK-8测定和Caspase-3ELISA测定分别测量细胞迁移,增殖和凋亡。用Western blot检测敲除GACAT3后p21,Bax和E-cadherin的表达变化。结果表明,与配对的正常组织相比,膀胱癌组织中的GACAT3上调。抑制细胞增殖,在靶向GACAT3的CRISPR-Cas13转染的T24和5637细胞系中观察到细胞凋亡增加,运动性降低。GACAT3的下调增加了p21,Bax和E-cadherin的表达,使这些基因沉默可以消除GACAT3敲低诱导的表型变化。还揭示了GACAT3的ceRNA机制。通过使用CRISPR-Cas13生物技术,我们建议GACAT3可能是诊断和治疗膀胱癌的新靶标。

更新日期:2021-01-18
down
wechat
bug