Knockdown of Decoy Receptor 3 Impairs Growth and Invasiveness of Hepatocellular Carcinoma Cell Line of HepG2

Background：Decoy receptor 3 （DcR3） binds to Fas ligand （FasL） and inhibits FasL-induced apoptosis.The receptor is overexpressed in hepatocellular carcinoma （HCC）,and it is associated with the growth and metastatic spread of tumors.DcR3 holds promises as a new target for the treatment of HCC,but little is known regarding the molecular mechanisms underlying the oncogenic properties of DcR3.The present work,therefore,examined the role of DcR3 in regulating the growth and invasive property of liver cancer cell HepG2.Methods：HepG2 cells were stably transfected with lentivirus-based short hairpin RNA vector targeting DcR3.After the knockdown of DcR3 was confirmed,cell proliferation,clone formation,ability of migrating across transwell membrane,and wound healing were assessed in vitro.Matrix metalloproteinase-9 （MMP 9） and vascular epithelial growth factor （VEGF）-C and D expressions of the DcR3 knockdown were also studied.Comparisons between multiple groups were done using one-way analysis of variance （ANOVA）,while pairwise comparisons were performed using Student＇s t test.P 〈 0.05 was regarded statistically significant.Results：DcR3 was overexpressed in HepG2 compared to other HCC cell lines and normal hepatocyte Lo-2.Stable knockdown of DcR3 slowed down the growth of HepG2 （P 〈 0.05） and reduced the number of clones formed by 50% compared to those without DcR3 knockdown （P 〈 0.05）.The knockdown also reduced the migration of HepG2 across transwell matrix membrane by five folds compared to the control （P 〈 0.05） and suppressed the closure of scratch wound （P 〈 0.05）.In addition,the messenger RNA levels of MMP 9,VEGF-C,and VEGF-D were significantly suppressed by DcR3 knockdown by 90% when compared with the mock control （P 〈 0.05）.Conclusions：Loss of DcR3 impaired the growth and invasive property of HCC cell line of HepG2.Targeting DcR3 may be a potential therapeutic approach for the treatment of HCC.

Effect of Liuweidihuang pill and Jinkuishenqi pill on inhibition of spontaneous breast carcinoma growth in mice

OBJECTIVE: To investigate the preventing and treating action of Liuweidihuang pill(LP) and Jinkuishenqi pill(JP) on spontaneous breast carcinoma in mice.METHODS: A model of spontaneous breast carcinoma was derived from 11.5-month-old female Kunming breeding mice following the delivery of several litters. The mice were randomly divided into five groups: model control group(C),Liuweidihuang pill high-dose group(LH; 4.6 g·kg1·d1),Liuweidihuang pill low-dose group(LL;2.3 g·kg1·d﹣1),Jinkuishenqi pill high-dose group(JH; 4.6 g·kg﹣1·d1) and Jinkuishenqi pill low-dose group(JL;2.3 g·kg﹣1·d﹣1). Cancer tissue volume was measured by water immersion. Histopathology was analyzed by hematoxylin and eosin staining. Vascular endothelial growth factor(VEGF),extracellular signal-regulated kinase(ERK) and cyclin D1 protein expression in cancer tissue was assayed by western blotting.RESULTS: Compared with the control group,cancer tissue volume and weight were lower in the LP and JP groups,and survival time was longer. The expression of VEGF,ERK and Cyclin D1 were inhibited in the LP and JP groups(P < 0.05),and cell differentiation was increased. Tumor weights and volumes and VEGF,ERK and Cyclin D1 expression in LL or LH were significantly lower than in JL and JH(P < 0.01).CONCLUSION: Both LP and JP could restrain cancer growth and promote cancer cell differentiation;moreover,LP was more effective than JP The likely mechanism of action was via inhibition of VEGF,ERK and cyclin D1.