Fig. 4.

CB₂ high expression represented a better prognosis of HCC patients and CB2 knockdown reversed the inhibition of HCC cell proliferation induced by MDA19. a Kaplan-Meier plotter (http://kmplot.com/analysis/) is a bioinformatics website that assesses the effect of 54,675 genes on survival using 10,461 cancer samples. The survival curves of HCC patients with high or low CB₂ mRNA level were acquired by searching on Kaplan-Meier plotter; (Affy id/Gene symbol: 1269_s_at; Survival: RFS (n = 364); Follow up threshold: all; and Auto select best cutoff and user selected probe set were selected to generate these figures); (b) The correlation of CB₂ expression with clinical factors of HCC patients; (c) Three siRNAs targeting CB₂ were synthesized and introduced into Hep3B and HepG2 cells. The mRNA expression of CB₂ was detected by qRT-PCR. d CB₂ was knocked down by using RNAi technology and western blot was used to detect the interference efficiency on protein level; (e) CCK8 was used to detect cell proliferation of HCC cells in NC (negative control), CB₂-KD and MDA19 + CB₂-KD groups. NC: HCC cells were transfected with siNC (50nM); CB₂-KD: HCC cells were transfected with CB₂ siRNA (50nM); MDA19 + CB₂-KD: HCC cells were transfected with CB2 siRNA (50nM) and treated with MDA19 (30μM for Hep3B and 40μM for HepG2). All experiments were performed at 3 times. *P < 0.05