Fig. 1.

NORAD enhanced the proliferation and metastasis of PCa cells

a, The expression of NORAD in 74 PCa tissues and adjacent normal tissue was detected by qRT-PCR. * p < 0.05, ** p < 0.01, vs. adjacent normal tissue. b, The expression of NORAD in 25 non-bone metastatic PCa tissues and 49 bone metastasis PCa tissues among 74 PCa patients was detected by qRT-PCR. ** p < 0.01, vs. non-bone metastatic PCa tissues. c, Kaplan-meier curves of PCa survival (high NORAD, n = 37; low NORAD, n = 37). d, The expression of NORAD in PCa cell lines (22Rv1, C4-2B, Du145, and PC-3) and normal prostate epithelial cell line RWPE-1 was detected by qRT-PCR and the cells with the highest and lowest NORAD expression were selected for follow-up studies. * p < 0.05, ** p < 0.01, vs. RWPE-1 cells. e, The knockdown efficiency of sh-NORAD against NORAD in PC-3 cells was detected by qRT-PCR, and the most efficient sequence was selected for subsequent experiments. ** p < 0.01, vs. PC-3 cells treated with sh-NC. f, The overexpression efficiency of OE-NORAD against NORAD in 22Rv1 cells was detected by qRT-PCR. ** p < 0.01, vs. PC-3 cells treated with vector. g, CCK-8 assay of proliferation of PCa cells upon treatment with sh-NORAD or sh-NC. * p < 0.05. h, Representative images of Transwell assay of migration and invasion of PCa cells upon treatment with sh-NORAD, OE-NORAD or sh-NC. * p < 0.05, ** p < 0.01, vs. PC-3 cells treated with sh-NC or 22Rv1 cells treated with vector. i, The effect of NORAD on the migration of PCa cells towards HS-5-CM was detected by Transwell. ** p < 0.01, vs. PC-3 cells treated with sh-NC or 22Rv1 cells treated with vector. The measurement data were expressed as mean ± standard deviation. Paired t-test was used for comparison between cancer tissue and adjacent normal tissue; unpaired t-test was used for other two groups; ANOVA was used for comparison among multiple groups with Tukey’s post-hoc test; Comparisons between two groups with different time points were performed using two-way ANOVA followed by Bonferroni. The Kaplan-Meier and log-rank test were used to analyze the survival rates. The cell experiment was repeated 3 times