Knockout of EGFL6 by CRISPR/Cas9 Mediated Inhibition of Tumor Angiogenesis in Ovarian Cancer.

Qi Chen,Simin Zhang,Yinmei Zhang,Chunyan Liu,Tongyi Lu,Wenhui Zhu,Ning Deng

doi:10.3389/fonc.2020.01451

Qi Chen, Simin Zhang + Show 5 more

Open Access

https://doi.org/10.3389/fonc.2020.01451

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Abstract

Tumor angiogenesis plays an important role in the progression and metastasis of ovarian cancer. EGFL6 protein is highly expressed in ovarian cancer and has been proposed to play an important role in promoting tumor angiogenesis. Here, a CRISPR/Cas9 system was used to knockout the EGFL6 gene in the ovarian cancer cell line SKOV3, using specific guide RNA targeting the exons of EGFL6. The knockout of EGFL6 markedly inhibited the proliferation, migration, and invasion of SKOV3 cells, as well as promoted apoptosis of tumor cells. In the nude mouse model of ovarian cancer, knockout of EGFL6 remarkably inhibited tumor growth and angiogenesis. The transcript profile assays detected 4,220 differentially expressed genes in the knockout cells, including 87 genes that were correlated to proliferation, migration, invasion, and angiogenesis. Moreover, Western blotting confirmed that EGFL6 knockout downregulated the FGF-2/PDGFB signaling pathway. Thus, the results of this study indicated that EGFL6 could regulate cell proliferation, migration, and angiogenesis in ovarian cancer cells by regulating the FGF-2/PDGFB signaling pathway.

Highlights

Ovarian cancer is a kind of malignant tumor that poses serious threat to women’s health and is one of the most lethal gynecological tumors [1, 2]
The expression of Epidermal growth factor-like domain multiple 6 (EGFL6) was detected in a panel of human ovarian cancer cells (CAOV3, COV-362, COV-504, EFO-27, ES2, OV-90, SKOV3, and TOV-21G), and EGFL6 expression in human umbilical vein endothelial cells (HUVECs) served as a negative control (Figure 1B)
The results showed that EGFL6 was expressed in all the ovarian cancer cell lines but with variations

Summary

INTRODUCTION

Ovarian cancer is a kind of malignant tumor that poses serious threat to women’s health and is one of the most lethal gynecological tumors [1, 2]. The angiogenic factors of ovarian cancer can be targeted for inhibiting tumor growth and metastasis. The murine EGFL6 promotes the migration of porcine vascular endothelial cells (SVEC) through the activation of the ERK signaling pathway and its downstream effector molecules, resulting in angiogenesis [17,18,19,20]. Considering the multiple mechanisms proposed in different cancers, the role of EGFL6 in tumor angiogenesis and progression of ovarian cancer requires further investigation. The EGFL6 was knocked out by CRISPR/Cas in an ovarian cancer cell line, using a specific guide RNA (gRNA) designed to target the exons of EGFL6. The role of EGFL6 in the proliferation and migration of cells, tumor growth, and angiogenesis in ovarian cancer was assayed. The molecular mechanism of EGFL6 function in the tumorigenesis of ovarian cancer needs further investigation

MATERIALS AND METHODS

RESULTS

DISCUSSION

ETHICS STATEMENT