Long non-coding RNA AGAP2-AS1 affects biological behavior of colon cancer cells through the YAP signaling pathway / 中国肿瘤临床

ABSTRACT

To investigate the expression of AGAP2-AS1 in colon cancer, its influence on the malignant behavior of colon cancer cells, and its effects on the Yes-associated protein (YAP) signaling pathway. Methods: The expression of AGAP2-AS1 in 457 colon cancer samples and 42 healthy samples was obtained from TCGA. Twenty matched colon cancer and adjacent cancer tissues were collected from patients diagnosed in Yiwu Central Hospital and confirmed by pathology department from January 2018 to March 2019. The expression of AGAP2-AS1 in these tissues, as well as in SW480, HCT-116, and NCM460 cells, was detected by real-time fluorescent quantitative PCR. The changes in cell proliferation, migration and apoptosis were analyzed and detected using a CCK-8 kit, and clone formation, wound-healing assay, and flow cytometry experiments. The levels of YAP, p-YAP, matrix metalloproteinase-2 (MMP2), and MMP9 were evaluated by Western blot. Immunofluorescence was used to examine the localization of YAP in HCT-116 cells. The levels of YAP, p-YAP, MMP2, and MMP9 proteins were evaluated after the co-transfection of pcDNA3.1-AGAP2-AS1 and si-YAP plasmids. Results: TCGA indicated that the expression of AGAP2-AS1 was significantly increased in colon cancer tissues. Similarly, in the present study, AGAP2-AS1 expression was significantly increased in colon cancer tissues and cells. After AGAP2-AS1 knockdown/overexpression, the capacity of migration and proliferation was significantly reduced/increased, and apoptosis was significantly increased/inhibited in colon cancer cells. Moreover, AGAP2-AS1 knockdown triggered the phosphorylation of YAP, and a significant reduction in the expression of MMP2 and MMP9 (P<0.05). Co-transfection studies revealed that AGAP2-AS1 upregulated the expression of MMP2 and MMP9 via activation of the YAP pathway (P<0.05). Conclusions: AGAP2-AS1 was highly expressed in colon cancer tissues and cell lines. AGAP2- AS1 can induce proliferation and migration, and inhibit apoptosis in colon cancer cells. In addition, AGAP2- AS1 regulated MMP2 and MMP9 expression by activating the YAP pathway, thereby affecting colon cancer metastasis.