Construction of NFIB Knock-Out Bladder Cancer Cell Line by CRISPR-Cas9 Technology and Its Transcriptomics Analysis
SHEN Lu1, SUN Yue1, WU Yanqian2, JIANG Ying2, HONG Yeting2, LYU Jianxin1,2*
In order to explore the biological function of NFIB gene in bladder cancer, CRISPR-Cas9 technology was utilized to knock out NFIB gene in bladder cancer cell line, and transcriptomics analysis was used to detect the DEGs (differentially expressed genes) between control cell NFIB-NC and NFIB knock out cells NFIBKO-I and NFIB-KO-II, and KEGG enrichment analysis was performed. The results of transcriptomics analysis were also verified by Western blot and qRT-PCR. Compared with the control cell NFIB-NC, there were 134 DEGs in NFIB knock-out cell line NFIB-KO-I, including 62 upregulated genes and 72 downregulated genes, and 131 DEGs in NFIB knock-out cell line NFIB-KO-II, including 50 upregulated genes and 81 downregulated genes. The results of KEGG enrichment analysis showed that DEGs were tightly related to PI3K-AKT signaling pathway, and the results of Western blot confirmed that the expression level of p-AKT was significantly upregulated in bladder cancer cell after knocking out NFIB. The results of qRT-PCR showed that after knocking out NFIB, the expression level of ITGA4 was upregulated, and the levels of TNC and ANGPT4 were downregulated. The study revealed the regulated molecular signaling pathways of NFIB, which could provide a basis for the follow-up research on the molecularmechanism of NFIB mediating the occurrence and development of bladder cancer.
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