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Establishment and characterization of A549 tumor monoclonal cell line with UCHL1 gene deletion

ZHANG Yu¹, SHI Yan², SONG Di¹, ZHANG Xuan², YU Lin², HE Ya-Ping², SUN Zhao-Gui^2,*

¹Shanghai Medical College, Fudan University, Shanghai 200032, China；²National Population and Family Planning Key Laboratory of Contraceptive Drugs & Devices, Shanghai Institute of Planned Parenthood Research, Shanghai 200032, China

Abstract

The purpose of this study was to investigate the effects of ubiquitin C-terminal hydrolase-L1 (UCHL1) on non-small cell lung cancer cell line A549. UCHL1 gene knockout A549 cell line was constructed by CRISPR-CAS9 gene editing technique. The mRNA and protein levels of UCHL1 were examined by RT-PCR and Western blot, respectively. Cell proliferation and cycles were analyzed by CCK-8 method and flow cytometry, respectively. The sensitivity of A549 cells to cisplatin was detected by CCK-8 method. Migration ability of A549 cells was detected by scratch assay and Transwell test, and p-Erk expression level was assessed by Western blot. The results showed that UCHL1 gene knockout A549 cells were successfully constructed by CRISPR-CAS9 gene editing technique. After UCHL1 gene knockout, there was no significant change in cell proliferation and cell cycle ratios in A549 cells. UCHL1 gene knockout A549 cells exhibited decreased sensitivity to cisplatin and migration activity, as well as increased p-Erk expression level. These results suggest that the loss of UCHL1 gene function may reduce the sensitivity and migration ability of A549 cells, and this effect may be related to the activation of Erk1/2 signaling pathway.

Key words: UCHL1; CRISPR-CAS9; non-small cell lung cancer; drug resistance; migration

Received: 2017-09-26　　Accepted: 2018-03-30

Corresponding author: 孙兆贵　　E-mail: sunzgbio@126.com

DOI: 10.13294/j.aps.2018.0032

Citing This Article：

ZHANG Yu, SHI Yan, SONG Di, ZHANG Xuan, YU Lin, HE Ya-Ping, SUN Zhao-Gui. Establishment and characterization of A549 tumor monoclonal cell line with UCHL1 gene deletion. Acta Physiol Sin 2018; 70 (2): 184-192 (in Chinese with English abstract).