Aberration of lncRNA LINC00460 is a Promising Prognosis Factor and Associated with Progression of Clear Cell Renal Cell Carcinoma

Figures & data

Table 1 Clinical Characteristics of ccRCC Patients and Their Association with LINC00460 Expression

Characteristics	Cases n = 113	LINC00460 Expression		P values
		Low (n = 54)	High (n = 59)
Gender
Male	72	34	38	0.873
Female	41	20	21
Age
≤ 55	59	30	29	0.496
> 55	54	24	30
Tumor size (cm)
≤ 5	63	32	31	0.473
> 5	50	22	28
Lymph node metastasis
Negative	79	44	35	0.010
Positive	34	10	24
Histological grade
I–II	86	41	45	0.966
III–IV	27	13	14
TNM stage
I–II	76	42	34	0.023
III–IV	37	12	25

Note: Differences were analyzed using Pearson’s chi-square test.

Figure 1 Relative LINC00460 expression in ccRCC tissues and cell lines. (A) The qRT-PCR analysis detected the expression level of LINC00460 in the RCC tissues and the corresponding normal tissues. (B) qRT-PCR analysis of LINC00460 in ccRCC cell lines and normal renal epithelial HK-2 cells. **P < 0.01, ***P < 0.001.

Table 2 Multivariate Cox Analyses of Overall Survival of ccRCC Patients

Characteristics	Multivariate Cox Analysis
	HR	95% CI	P value
LINC00460	2.619	1.110–6.178	0.028
Gender	1.450	0.645–3.259	0.369
Age	1.835	0.842–3.998	0.127
Tumor size	1.423	0.647–3.131	0.381
Lymph node metastasis	2.467	1.088–5.591	0.031
Histological grade	1.614	0.627–4.155	0.321
TNM stage	2.577	1.039–6.391	0.041

Note: Data are analyzed using multivariate Cox regression analysis.

Figure 2 Kaplan–Meier curves for ccRCC patients with high and low levels of LINC00460 (Log rank test: P = 0.014).

Figure 3 Knockdown of LINC00460 inhibited the proliferation, migration and invasion of 786-O and Caki-1 cells. (A and B) si-NC or si-LINC00460 were transfected into 786-O and Caki-1 cells, and transfection efficiency was verified by qRT-PCR. (C and D) Cell proliferation of 786-O and Caki-1 was determined after transfection with si-NC or si-LINC00460 by CCK-8 assays. ***P or ****P < 0.001.

Figure 4 (A and B) The migration abilities of 786-O and Caki-1 cells were accessed after transfection via transwell migration assays. (C and D) The invasive abilities of 786-O and Caki-1 cells were tested after transfection using transwell invasion assays. ***P < 0.001.

Figure 5 MiR-149-5p acted as the target of LINC00460. (A) The binding sites between LINC00460 and miR-149-5p. (B) The qRT-PCR analysis detected the expression level of miR-149-5p in the RCC tissues and the corresponding normal tissues. (C) qRT-PCR analysis of miR-149-5p in ccRCC cell lines and normal renal epithelial HK-2 cells. (D) The correlation between LINC00460 and miR-149-5p in ccRCC tissues was analyzed using Spearman correlation analysis. (E) Luciferase activity was examined in ccRCC cells co-transfected with WT-LINC00460 or MUT-LINC00460 reporter plasmid and miR-149-5p or miR-NC. (F) RIP assay indicated that both LINC00460 and miR-149-5p were significantly enriched in Ago2 immunoprecipitate. Empty vector was used as a nonspecific control. IgG acted as an internal control. *P < 0.05, **P < 0.01, ***P < 0.001.