Objective To investigate the effect of CD44 regulation of HIF-1α/PLOD2 signaling pathway on epithelial mesenchymal transformation in clear cell renal cell carcinoma (ccRCC). Methods The expression of CD44 in ccRCC tissues was analyzed by bioinformatics database. The expression of CD44 in ccRCC cells was detected by qRT-PCR. The expressions of epithelial- mesenchymal transition, metastasis-related proteins, and HIF-1α and PLOD2 were analyzed by Western blot. Cell proliferation, migration and invasion were detected by clonal formation assay and transwell assay respectively. Results D44 was significantly upregulated in ccRCC cells. Compared with shNT group, cell proliferation, migration and invasion ability of sh-CD44 group were significantly decreased, the expression of E-cadherin protein was increased, and the expressions of N-cadherin, MMP-2 and MMP-9 protein were decreased. Compared with the Vector group, the proliferation, migration and invasion of cells in the CD44-OE group were significantly enhanced, the expression of E-cadherin protein was decreased, and the expressions of N-cadherin, MMP-2 and MMP-9 protein were increased. In addition, the expressions of HIF-1α and PLOD2 were decreased in sh-CD44 group compared with shNT group. Compared with Vector group, the expressions of HIF-1α and PLOD2 in CD44-OE group were increased. The mRNA level of HIF-1α in the sh-CD44 group did not change, but VHL bound to HIF-1α increased, which promoted the degradation of HIF-1α protein. Compared with the Vector group, the promoter activity of PLOD2 in the CD44-OE group was enhanced. In addition, compared with Vector+DMSO group, the proliferation, migration and invasion of cells in CD44-OE+DMSO group were significantly improved. Compared with the CD44-OE+DMSO group, the proliferation, migration and invasion of cells in the CD44-OE+ PX-478 group were significantly reduced. In addition, compared with Vector+DMSO group, the expression of E-cadherin protein decreased, while the expressions of N-cadherin, MMP-2, MMP-9, HIF-1α and PLOD2 protein were increased in CD44-OE+DMSO group. Compared with CD44-OE+DMSO group, the expression of E-cadherin protein was increased in CD44-OE+PX-478 group, while the expressions of N-cadherin, MMP-2 and MMP-9, HIF-1α and PLOD2 protein were decreased. Conclusion The results of this study indicate the roles of CD44 in promoting the proliferation and metastasis of ccRCC cells. These results suggest that targeting CD44 or HIF-1α/PLOD2 pathways may be new targets for ccRCC therapy.
Key words
CD44 /
HIF-1α/PLOD2 pathway /
clear cell renal cell carcinoma /
epithelial-mesenchymal transition
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