Fig. 6

SNHG1 regulates expression of the miR-154-5p target gene, CCND2. a CCND2 expression was detected by qRT-PCR in SNHG1 siRNAs transfected or SNHG1 siRNAs and miR-154-5p inhibitors co-transfected HCT-116 cells. b CCND2 expression was detected by qRT-PCR in SNHG1 vector transfected or SNHG1 vector and miR-154-5p mimics co-transfected HCT-116 cells. c Western blot analyses of CCND2 expression after knockdown of SNHG1, overexpression of miR-154-5p or knockdown of SNHG1 + inhibition of miR-154-5p in HCT-116 cells. d Dual luciferase reporter assays demonstrated that miR-154-5p overexpression reduced Luc-CCND2 luciferase activity and SNHG1 overexpression abolished miR-154-5p induced reductions in luciferase activity in HCT-116 cells. e CCND2 expression was measured by western blot after silencing of endogenous SNHG1 and transfection with either SNHG1-mut vector, which contains mutations at the putative miR-154-5p binding site, or SNHG1 vector in HCT-116 cells. f The correlation between CCND2 and SNHG1 expression analyzed in 30 paired colorectal cancer samples (n = 30, r = 0.38, P = 0.036). g EdU assays demonstrated HCT-116 cells proliferation rates after knockdown of SNHG1, knockdown of CCND2 or both knockdown of SNHG1and CCND2. h CCK-8 assays demonstrated that CCND2 knockdown could reverse growth promotion caused by SNHG1 overexpression in HCT-116 cells. i Immunohistochemistry analysis of CCND2 protein levels in tumor tissues formed from SNHG1 knockdown or control cells. j Detection of CCND2 protein levels in colorectal cancer and normal tissues by IHC. Scale bar = 50 μm. *P < 0.05, **P < 0.01 and ***P < 0.001