ROCK1基因敲除对乳腺癌细胞系迁移及侵袭的影响

doi:10.3969/j.issn.1004-616x.2019.03.004

Abstract

Abstract: OBJECTIVE: To construct a ROCK1 gene-knockout model using the CRISPR/Cas9 system and to investigate effects of the ROCK1 knockout on MCF-7 breast cancer cell migration and invasion. METHODS: Three pairs of 20 bp sgRNA targeting ROCK1 were chemically synthesized and inserted into CRISPR expression vectors. The ROCK1 knockout cells were selected with lentivirus infection of MCF-7 cells, and were identified by gene sequencing and Western blot. Effects of the knockout on MCF-7 cell migration and invasion were evaluated by wound-healing and Transwell assays. RESULTS:Western blot data show that ROCK1 knockout MCF-7 cells were successfully and stably established by usage of the CRISPR/Cas9 system. Wound-healing assays show that the wound closure of ROCK1 knockout cells was significantly lower than that of their parental cells[(60.600±0.047)% and (80.404±0.018)%,respectively;P=0.003]. Transwell assays without matrigel show that the migration capability of ROCK1 knockout cells was significantly decreased compared with the control group(271.3 ±5.0 and 448.3 ±5.5, respectively; P=0.000) by 24 h. Transwell assays with matrigel show the invasion capability of ROCK1 knockout cells at 48 h was deeply suppressed compared to that of the controls (1.7±2.9 and 298.3±5.7,respectively;P=0.000). CONCLUSION:Knockout of ROCK1 gene caused significant inhibition of the migration and invasion ability of breast cancer cells,suggesting that ROCK1 may play an important role in the invasion and metastasis of breast cancer.

Key words: Rho-associated protein kinase 1, CRISPER/Cas9, migration, invasion

CLC Number:

WANG Xiaojie, TENG Yu, GU Meng, WANG Ziyu, ZHAO Xiaoting, YUE Wentao. Effects of ROCK1 knockout on breast cancer cell migration and invasion[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2019, 31(3): 193-197,202.

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Effects of ROCK1 knockout on breast cancer cell migration and invasion

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