| Objective: To investigate the effect of microRNA-378(miR-378)on cell proliferation and migration in renal cancer cell lines and to explore its possible mechanism in vitro.And To investigate the effect of microRNA-378(miR-378)on cell proliferation in prostate cancer cell in vitro and its effect on cell differentiation in preosteoblast cell.Methods: In Renal cancer,miR-378 expression was analyzed using datasets from The Cancer Genome Atlas(TCGA)and confirmed by quantitative real-time fluorescent PCR(qRT-PCR)in human renal carcinoma cell lines and human renal proximal tubular epithelial cell line HK2.MiR-378 mimic and inhibitor were synthesized and then transfected into 786-O and Caki-1 cells.Proliferation of 786-O and Caki-1 cells was detected by CCK-8 assay.Migration of Caki-1 cells was examined by Transwell and wound healing assays,respectively.Potential target genes of miR-378 were screened by bioinformatics and confirmed by qRT-PCR and western blot in renal cancer cell lines,respectively.In prostate cancer,the morphology and purity of exosome collectecd by ultracentrifugation or total exosome isolation reagent were analyzed by electron microscopy and particle size analysis.MiR-378 mimic were synthesized and then transfected into prostate cancer cell line LNCaP cells and pre-osteoblast cell line MC3T3-E1 cells respectively.Proliferation of LNCaP cells was detected by CCK-8 assay.The expression of miR-378 in LNCaP or BPH1 culture medium supernatant-derived exosome was detected by qRT-PCR.Potential target genes of miR-378 in LNCaP and MC3T3-E1 were screened by bioinformatics.The expression of miR-378 target genes was detected by qRT-PCR and immunofluorescence(IF).Results: The expression level of miR-378 in renal cancer cell lines was significantly upregulated compared to that in HK2 cell(P<0.001).After transfecting with miR-378 mimic,both proliferation(P<0.01)and migration(P <0.001)of renal cancer cells were remarkably increased.In contrast,after transfecting with miR-378 inhibitors,proliferation(P<0.01)and migration(P <0.001)of renal cancer cells were remarkably suppressed.By bioinformatics assay,large tumor suppressor 2(LATS2)was identified as a target gene of miR-378 in renal cancer.The mRNA and protein expression of LATS2 were negatively related with expression of miR-378(P<0.01)in renal cancer cell lines by qRT-PCR and western blot.In prostate cancer,it was shown by the results of qRT-PCR that the expression of miR-378 in exosome from LNCaP culture medium supernatant was significantly upregulated compared to that from BPH1 culture medium supernatant(P<0.01).Overexpression of miR-378 and inhibition of exosome release simultaneously inhibited the proliferation of LNCaP cells in vitro(P < 0.01)and upregulated expression of osteocalcin and OSX,markers for labeling differentiation of pre-osteoblast cells.By bioinformatics assay,Monoamine oxidaseA(MAOA)was identified as a target gene of miR-378 in prostate cancer cells,and GLI family zinc finger 3(Gli3)and human immunodeficiency virus type I enhancer binding protein 3(HIVEP3,also named Schnurri3)were also identified as target genes of miR-378 in MC3T3-E1 cells.It was verified by the results of qRT-PCR that expression of target genes was negatively correlated with miR-378 expression.Overexpressing miR-378 and inhibiting exosome release simultaneously in LNCaP cells significantly inhibited the expression of MAOA and Shh,a downstream target gene of MAOA(P < 0.01).Overexpressing miR-378 and inhibiting exosome release simultaneously in MC3T3-E1 cells significantly inhibited expression of miR-378 target genes Gli3 and Schnurri3(P < 0.01)..Conclusion: In renal cancer,overexpression of miR-378 promotes proliferation and migration of renal cancer cells in vitro by targeting LATS2.In prostate cancer,overexpression of miR-378 in vitro can inhibit proliferation of prostate cancer cells by inhibiting the expression of MAOA,and can promote differentiation of pre-osteoblast cells by inhibiting Gli3 and Schnurri3 expression. |
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