The ratio of the relative migration in miR-182-5p inhibitor group was elevated by 1.51 times in 786-O (P?=?0.001) and raised by1.58 times in Caki-1 (P?=?0.005) (Fig. we found that UCA1 was significantly up-regulated in renal cancer. Moreover, increased UCA1 expression was positively correlated with differentiation and advanced TNM stage. Further experiments demonstrated that knockdown of UCA1 inhibited malignant phenotypes and Notch signal path of renal cancer cells, and miR-182-5p was reverse function as UCA1. UCA1 functioned as a miRNA sponge to positively regulate the expression of Delta-like ligand 4(DLL4) through sponging miR-182-5p and subsequently promoted malignant phenotypes of renal cancer cells, thus UCA1 playing an oncogenic role and miR-182-5p as an antioncogenic one in renal cancer pathogenesis. Conclusion UCA1-miR-182-5p-DLL4 axis is involved in proliferation and progression of renal cancer. Thus, this study demonstrated that UCA1 plays a critical regulatory role in renal cancer cell and UCA1 may serve as a potential diagnostic biomarker and therapeutic target of renal cancer. value of less than 0.05 was considered to be statistically significant. Results Up-regulation of UCA1 and low-expression of miR-182-5p in renal cancer tissues, cells and both correlation with clinical pathologic factors The relative expression level of UCA1 and Cytarabine miR-182-5p was detected by using Real-Time qPCR in a total of 88 patients with renal cancer. Compared to matched normal peritumoral tissues, the UCA1 expression was up-regulated remarkably in 68.2% (60 of 88) of cancer tissues (valuevalue
Gender?Male4711 (23.4%)36 (76.6%)0.474?Female4113 (31.7)28 (68.3%)Tumor size (cm)???7?cm5016 (32.0%)34 (68.0%)0.335?>7?cm388 (21.1%)30 (78.9%)Age? 554315 (34.9%)28 (65.1%)0.152??>?55459 (20.0%)36 (80.0%)Differentiation?Moderate/poor508 (16.0%)42 (84.0%)0.008**?Well3816 (42.1%)22((57.9%)TNM stage?T0C12612 (11.5%)14 (88.5%)0.017*?T2C46212 (38.7%)50 (61.3%)Lymph node metastasis(N)?N07921 (26.6%)58 (73.4%)0.700?N1 or above93 (33.3%)6 (66.7%) Open in a separate window (*P?0.05, **P?0.01) TNM according to staging TNM of American Joint Committee on Cancer (AJCC) in 2010 2010 Knockdown of UCA1 and up-regulation of miR-182-5p inhibited cell Cytarabine proliferation of renal cell lines. Up-regulation of UCA1 and down-regulation of mi-182-5p promoted cell proliferation of renal cell lines We further determined whether UCA1 promotes cell proliferation and miR-182-5p restrained cell proliferation in renal cancer. The relative expression level of UCA1 and miR-182-5p were analyzed by qRT-PCR at 48?h after transfection of shRNA, miRNA mimics or inhibitor in in 786-O and Caki-1 cell lines, and after transfection of pcDNA3.1-UCA1 in 293?T and RPTEC cell line. The relative expression levels of UCA1 was decreased by 48.17% in 786-O (P?=?0.007) and was decreased by 43.84% in Caki-1(P?=?0.011) cells were down-regulated significantly by shUCA1 at 48?h post transfection (Fig. ?(Fig.2a).2a). And the relative expression levels of UCA1 was up-regulated significantly in by 3.99 times in 293?T cells (P?0.001) at 48?h post transfection of pcDNA3.1-UCA1 (Fig. ?(Fig.2b).2b). And the relative expression levels of UCA1 was up-regulated significantly in by 4.026 times in RPTEC cells (P?0.001) at 48?h post transfection of pcDNA3.1-UCA1 (Fig. ?(Fig.22 c). And the relative expression levels of miR-182-5p were down-regulated significantly by 80.74% in 786-O (P?0.001) and by 73.75% in Caki-1(P?0.001) cells at 48?h Cytarabine post transfection of miR-182-5p inhibitor (Fig. ?(Fig.3a).3a). And the relative expression levels of miR-182-5p were up-regulated significantly in by 2.30 times in 786-O (P?0.001) and 2.21 times in Caki-1(P?0.001) cells at 48?h post transfection of miR-182-5p mimics (Fig. ?(Fig.33a). Open in a separate window Fig. 2 Knockdown and overexpression of UCA1 inhibited or promote cell proliferation. The relative expression level of UCA1 was significantly down-regulated by shUCA1 (a) and upregulated by pcDNA3.1-UCA1(b and c). ANOVA was used for the comparison of curves of cell proliferation. Cell proliferation was detected in both renal cancer cells after transfection of shRNA (d and e) and pcDNA3.1-UCA1 (f and g). Representative images of EdU assay and the relative fold changes of EdU positive cells were detected by shRNA (H and I) and pcDNA3.1-UCA1 (j and k). Assays were performed in triplicate, and data were shown as mean??standard deviation (SD) of CDK4 those biological replicates or samples (*P?0.05, **P?0.01) Open in a separate window Fig. 3 Knockdown and overexpression of miR-182-5p promote or inhibited cell proliferation. The relative expression level of miR-182-5p was significantly down-regulated by miR-182-5p inhibitor and up-regulated by miR-182-5p mimics (a). ANOVA was used for the comparison of curves of cell proliferation. Cell proliferation was detected in both renal.