[Rev-erbß Knock-out Affectsthe Proliferation, Migration and Invasion Ability of Hepatocellular Carcinoma HepG2 Cell Line in vitro].
Sichuan Da Xue Xue Bao Yi Xue Ban
; 50(4): 520-526, 2019 Jul.
Article
en Zh
| MEDLINE
| ID: mdl-31642229
ABSTRACT
OBJECTIVE:
To investigate the effect of nuclear receptor Rev-erbß knockout on proliferation and migration ability of human hepatocellular carcinoma cell line HepG2.METHODS:
-The Rev-erbß gene knockout HepG2 cell line was abtained by CRISPR/Cas9 genome editing technique with specific DNA modification of the target gene. The Rev-erbß gene targeting vectors were co-transfected into HepG2 cells. Through cloning and screening, the Rev-erbß gene knockout HepG2 cell line was constructed, PCR, sequencing and Western blot methods were carried out for the identification of the Rev-erbß gene knockout HepG2 cell line. The expression level of tumor migration and invasion-associated gene in Rev-erbß gene knockout cell was determined by real-time quantitative PCR (qRT-PCR) and was compared with normal cell as control.MTT, cell scratch and Transwell experiments were conducted in order to explore the effect of Rev-erbß gene on HepG2 cell's ability of proliferation, migration and invasion.RESULTS:
A Rev-erbß gene knockout monoclonal cell line, which was identified by PCR, sequencing and Western blot, was successfully constructed and named HepG2 C5 (Rev-erbß -/-). qRT-PCR results showed that Rev-erbß knockout resulted in up-regulation of matrix metalloproteinase-2 (MMP2), matrix metalloproteinase-9 (MMP9) and extracellular matrix protein-1 (ECM1) gene expression (P < 0.05) and down-regulation of E-cadherin (CDH1) gene expression (P=0.05).Results of MTT, cell scratch and transwell experiments showed that HepG2 C5 had stronger proliferation, migration and invasion ability than control cells (P < 0.05).CONCLUSION:
Rev-erbß gene knockout could change the expression of migration and adhesion-associated genes in HepG2 cell, and then affect the proliferation, migration and invasion ability of HepG2 cells.Palabras clave
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Base de datos:
MEDLINE
Asunto principal:
Proteínas Represoras
/
Movimiento Celular
/
Receptores Citoplasmáticos y Nucleares
/
Proliferación Celular
/
Invasividad Neoplásica
Tipo de estudio:
Prognostic_studies
Límite:
Humans
Idioma:
Zh
Revista:
Sichuan Da Xue Xue Bao Yi Xue Ban
Año:
2019
Tipo del documento:
Article
País de afiliación:
China