FSCN1 Promotes Glycolysis and Epithelial-Mesenchymal Transition in Prostate Cancer through a YAP/TAZ Signaling Pathway

Abstract

Objective: The aim of the study is to investigate the role and possible mechanism of fascin-1 (FSCN1) in the invasion, migration, glycolysis, and epithelial-mesenchymal transition (EMT) of prostate cancer.

Methods: Real-time quantitative polymerase chain reaction (qRT-PCR) was utilized to determine the mRNA expression level of FSCN1 in prostate cancer tissues and prostate cancer cells PC-3 and DU145. The transwell and the scratch test were applied to detect the invasion and migration abilities of cells, respectively. A metabolic assay was used for measuring the glucose consumption, lactate production, and the extracellular acidification rate (ECAR) in cells; western blot was used for checking FSCN1, EMT, and yes-associated protein/transcriptional co-activators with the PDZ-binding motif (YAP/TAZ) signaling pathway-related protein expression level in cells or tissues.

Results: FSCN1 was significantly highly expressed in prostate cancer tissues and cells. On the one hand, interference with the expression of FSCN1 could inhibit the invasion, migration, EMT, and glycolysis of prostate cancer cells. On the other hand, overexpression of FSCN1 promoted the invasion, migration, EMT, and glycolysis of prostate cancer cells. Besides, further mechanistic studies revealed that FSCN1 could activate the YAP/TAZ signaling pathway in prostate cancer cells.

Conclusion: FSCN1 promotes invasion, migration, EMT, and glycolysis in prostate cancer cells by activating the YAP/TAZ signaling pathway. FSCN1 may be used as a biomarker for the diagnosis or treatment in prostate cancer.

Figure 1

FSCN1 is highly expressed in prostate cancer tissues and cells. (a) qRT-PCR for detecting the mRNA level of FSCN1 in prostate cancer tissue (PCa tissue) and the corresponding adjacent tissue (adjacent tissue), ^∗∗P < 0.01; (b, c) western blot to detect the protein expression level of FSCN1 in the PCa group and the adjacent tissue group, and the relative protein expression level was analyzed, ^∗∗P < 0.01; (d) qRT-PCR to detect the mRNA expression level of FSCN1 in WPMY-1, PC-3, and DU145 cells; (e, f) western blot for the detection of the protein level of FSCN1 in WPMY-1, PC-3 and DU145 cells, and the relative protein expression level was analyzed. ^∗∗P < 0.01 vs. WPMY-1, ^##P < 0.01, and ^#P < 0.05 vs PC-3.

Figure 2

FSCN1 promotes prostate cancer cell invasion, migration, and EMT. (a) qRT-PCR for the detection of the mRNA expression of FSCN1 in PC-3 cells transfected with si-NC, si-FSCN1#1, and si-FSCN1#2, ^∗∗P < 0.01 vs si-NC; (b, c) western blot for the examination of the protein expression level of FSCN1 in PC-3 cells after si-NC, si-FSCN1#1, and si-FSCN1#2 transfection, ^∗∗P < 0.01 vs si-NC; (d) qRT-PCR to detect the mRNA expression level of FSCN1 in DU145 cells in groups of mock and FSCN1, ^∗∗P < 0.01; (e, f) western blot for determining the protein expression level of FSCN1 in DU145 cells in the mock group and the FSCN1 group; (g) transwell to measure the invasion ability of PC-3 cells in groups of si-NC, si-FSCN1#, and si-FSCN1#2 and DU145 cells in groups of mock and FSCN1; (h) scratch test for evaluating the migration ability of PC-3 cells in the si-NC group, si-FSCN1#1 group, and si-FSCN1#2 group and DU145 cells in the mock group and the FSCN1 group; (i, j) western blot to determine the protein expression level of E-cadherin, vimentin, and snail in PC-3 cells in the si-NC group, si-FSCN1#1 group, and si-FSCN1#2 group, ^∗∗P < 0.01 vs si-NC; (k, l) western blot for detecting the protein expression level of E-cadherin, vimentin, and snail in DU145 cells in the mock group and the FSCN1 group, ^∗∗P < 0.01.

Figure 3

FSCN1 promotes glycolysis in prostate cancer cells. (a–d) Determination of the glucose consumption (a), lactate production (b), ECAR (c), and extracellular pH (d) in PC-3 cells in the si-NC, si-FSCN1#1, and si-FSCN1#2 groups, ^∗P < 0.05 and ^∗∗P < 0.01 vs si-NC; E-H, assay of the glucose consumption (e), lactate production (f), ECAR (g), and extracellular pH in DU145 cells in the mock group and the FSCN1 group (h), ^∗∗P < 0.01.

Figure 4

FSCN1 activates the YAP/TAZ signaling pathway in prostate cancer cells. (a, b) Western blot measured the protein expression level of YAP, p-YAP, n-YAP, and TAZ in PC-3 cells in the si-NC, si-FSCN1#1, and si-FSCN1#2 groups, ^∗∗P < 0.01 vs si-NC; (c, d) western blot detected the protein expression level of YAP, p-YAP, n-YAP, and TAZ in DU145 cells in the mock group and the FSCN1 group, ^∗∗P < 0.01.