The Effect and Mechanism of miR-141-3p on Vasculogenic Mimicry in Human Prostate Cancer Cells by Targeted Regulating the KLF9 Expression
LIU Bide, WANG Shuheng, LI Xun, JIN Hongyong, ZHANG Xiao’an, DONG Qiang, LI Jiuzhi*
This study aimed to investigate the role of miR-141-3p in the VM (vasculogenic mimicry) of PCa (prostate cancer) cells and its mechanism. Normal human gastric epithelial cell lines (RWPE-1) and human PCa cell lines (PC-3, LNCaP and DU145) were cultured. The expression of miR-141-3p was detected using RT-qPCR. Western blot was used to detect the protein expression of KLF9. According to the expression difference of miR-141-3p and KLF9 in different PCa cell lines, as well as the purpose of the study, LNCaP and DU145 cell lines were selected for subsequent experiments. LNCaP and DU145 cells were divided into miR-141-3p control group, miR-141-3p inhibitor group, KLF9 overexpression group and KLF9 control group, respectively. miR-141-3p inhibitor was used to downregulate the expression of miR-141-3p, and KLF9 overexpression plasmid was used to upregulate the expression of KLF9. Vasculogenic mimicry was detected by Three-Dimensional culture. Cell proliferation activity was detected by CCK-8 assay. Wound-healing assay and Transwell assay were used to detect cell migration and invasion abilities. To evaluate the stemness of PCa cells, Western blot was conducted to detect the expression of stem cell markers, colony formation assay was used to evaluate cell colony formation ability, and flow cytometry was used to detect the proportion of CD133+ cells. Next, dual luciferase reporting assay and recue assay verified the targeting relationship between miR-141-3p and KLF9. The results showed that down-regulation of miR-141-3p or up-regulation of KLF9 could significantly inhibit the VM formation and stemness of PCa cells, and inhibit the proliferation, migration and invasion ability of PCa cells remarkably. The ability of miR-141-3p on promoting VM formation and stemness of PCa cells was achieved through targeting inhibit KLF9 expression. In conclusion, miR-141-3p regulates the VM formation ability of PCa cells by target regulating KLF9, which may be achieved by regulating the stemness of PCa cells.
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