Study On The Relationship Between Anti-proliferatiive Effect Of Oridonin On Human Colon Cancer Celline HCT116 And BMP7

ObjectiveTo study the relationship of anti-proliferative effect of ORI on human colon cancer cell-line HCT116 and BMP7,and the possible molecular mechanisms underlying the relationships.MethodsHCT116 cells were treated with ORI under the concentration of5μmol/L, 10μmol/L, and 15μmol/L respectively.1.Crystal violet staining, flow cytometry, PCR and Western Blot assay were introduced to detected the effect of oridonin on the proliferation, cell cycle and apoptosis in HCT116 cells.2. Xenograft in nude mice was used to study the effect of oridonin in colon cancer3. The effect of ORI on the expression of BMP7 and the endogenous expression in HCT116 cells was detected by PCR and Western Blot assay.4. BMP7 recombinant adenovirus(AdBMP7) and BMP7 antibody(anti-BMP7) were used to study the relationship between BMP7 andproliferation in HCT116 cells.5.Xenograft in nude mice was used to study the effect of AdBMP7 in colon cancer.6. The effect of ORI on Smad1/5/8, P-Smad1/5/8, p38 and p-p38 was detected by PCR and Western Blot assay.7. The relationship between p38 MAPK and proliferation in HCT116 were observed by introduce p38 MAPK inhibitor(SB).8. The anti-proliferative effect of BMP7 in HCT116 cells and its relationship with p38 MAPK was also dissected.Results1. ORI effectively inhibited the proliferation,induced cell cycle arrest at G2/M phase and apoptosis in HCT116 cells.2.ORI significantly inhibited tumor growth in nude mice3.ORI significantly up-regulate the expression of BMP7, BMP7 endogenous expression occurs in normal colonic epithelial cells and various colon cancer cell lines.4. In HCT16 cells, AdBMP7 can promote the expression of BMP7 and inhibit effect of ORI, while anti-BMP7 is represent a reversed effects as AdBMP7.5.AdBMP7 can inhibit the tumor growth and enhance anti-proliferative effect of ORI in nude mice.6.ORI exerted no significant effect on the level of Smad1/5/8,P-Smad1/5/8 and p38, but up-regulated the expression of p-p38.7.SB can promote the proliferation of HCT116 cells and reverse the inhibitory effect of ORI.8.AdBMP7 can promote the expression of p-p38, and enhance the effect of ORI on p-p38, while anti-BMP7 is reversed.Conclusions1. ORI inhibits the proliferation and induces apoptosis of HCT116 cells obviously, and arrests the cell cycle at the G2/M phase.2. ORI can up-regulate the expression of BMP7 and p-p38.3. ORI inhibits the proliferation and induces apoptosis by up-regulating BMP7 to activate the p38 MAPK signaling pathway.