Effects of IKBKE on Proliferation, Migration and Invasion of Pancreatic Cancer Cells by Regulating Hippo-YAP Signaling Pathway
ZHANG Zhenliang1*, HAO Jing2, ZHAO Xinglong3, MA Guangjun1, XU Bo1
This study aims to investigate whether IKBKE (inhibitor of nuclear factor kappa B kinase sub unit epsilon) can regulate Hippo-YAP (Yes-associated protein) signaling pathway to affect the proliferation, mi gration and invasion of pancreatic cancer cells. Pancreatic cancer and paracancer tissues were collected, IKBKE expression was detected by immunohistochemistry, and the relationship between IKBKE expression and clinico pathological features of patients was analyzed. The expression of IKBKE gene in pancreatic duct epithelial cells and pancreatic cancer cells was detected by qRT-PCR. Pancreatic cancer SW1990 cells were grouped into control group, si-NC group, si-IKBKE group, si-IKBKE+TDI-011536 (Hippo-YAP signaling pathway inhibitor) group, and Verteporfin group (Hippo-YAP signaling pathway activator). qRT-PCR was used to detect the expression level of IKBKE mRNA in each group. MTT assay and EdU staining were used to detect cell proliferation. Cell scratch assay was used to detect cell migration. Transwell was used to detect cell invasion. Flow cytometry was used to detect the apoptosis rate of cells. The expression levels of E-cadherin, N-cadherin, vimentin, cleaved caspase-3, PCNA (proliferating cell nuclear antigen), IKBKE, YAP, TAZ, p-YAP, p-TAZ, TEAD2 (TEA domain transcription factor 2), Axl (tyrosine protein kinase receptor), c-myc, Bcl-2 (B-cell lymphoblastoma-2), Bax (Bcl-2 associated X protein) proteins in cells were detected by Western blot. IKBKE was highly expressed in pancreatic cancer tissues, and its expression was positively correlated with tumor diameter, lymph node metastasis and differentiated status (P<0.05). Compared with pancreatic duct epithelial cells, the expression of IKBKE mRNA in several pancreatic cancer cells increased (P<0.05). Compared with the control group and si-NC group, the D490 (24 h, 48 h) values of SW1990 cells, proliferation rate, scratch healing rate, the number of cell invasions, and the expression levels of N cadherin, vimentin, PCNA, Bcl-2, YAP, TAZ, TEAD2, Axl, and c-myc proteins were decreased in the si-IKBKE group and Verteporfin group, the apoptosis rate, and the expression levels of E-cadherin, cleaved caspase-3, Bax, p YAP, and p-TAZ proteins were increased (P<0.05), the expression of IKBKE mRNA and IKBKE protein in the si IKBKE group decreased (P<0.05), while there was no statistically significant difference in the expression of IKBKE mRNA and IKBKE protein in the Verteporfin group (P>0.05). Compared with the si-IKBKE group, the D490 (24 h, 48 h) values of SW1990 cells, proliferation rate, scratch healing rate, the number of cell invasions, and the expres sion levels of N-cadherin, vimentin, PCNA, Bcl-2, YAP, TAZ, TEAD2, Axl, and c-myc proteins were increased in the si-IKBKE+TDI-011536 group, the apoptosis rate, and the expression levels of E-cadherin, cleaved caspase-3, Bax, p-YAP, and p-TAZ proteins were reduced (P<0.05), the expression of IKBKE protein showed no statistical significance (P>0.05), the expression levels of IKBKE mRNA and IKBKE protein in SW1990 cells of Verteporfin group were increased (P<0.05), the other indexes were not statistically significant (P>0.05). Downregulation of IK BKE expression may decrease the invasion, migration and proliferation of pancreatic cancer cells by up-regulating Hippo-YAP signaling pathway.
CN
EN