Study On The Relationship Between The Anti-proliferation Effect Of Resveratrol In LoVo Cells And BMP9/p38MAPK

Aim To study the effect of resveratrol on anti-proliferation in Lo Vo cells and the possible mechanism underlying this process.Methods We used crystal violet staining assay, flowcytometry assay, immunofluorescence staining, luciferase reporter assay, HE staining, RT-PCR, Western blot, and immunohistochemistry staining to analyze the effect of resveratrol on anti-proliferation in Lo Vo, so does the the possible mechanism. 1. Crystal violet staining, Western blot, RT-PCR and immunofluorescence staining were used to explore the effects of resveratrol on proliferation, apoptosis, and cell cycle in Lo Vo cells. 2. Xenograft tumor model was used tostudy the anti-tumor effects of resveratrol in vivo. 3. Luciferase reporter assay was used to detect the effects of resveratrol on BMP/Smad signaling pathway.4. With PCR and Western blot assay, we detected the effect of resveratrol on the m RNA and/or protein levels of BMP9, Smad1/5/8, p-Smad1/5/8, p38 MAPK, and p-p38 MAPK in Lo Vo cells. 5. Using recombinant adenovirus mediated BMP9 over-expression or knockdown to determine the role of BMP9 in resveratrol-induced proliferation inhibitory effect, and its relationship with p38 MAPK. 6. We detected the proliferation effects with p38 MAPK inhibitor and the effects of SB and resveratrol on the level of p38 MAPK, p-p38 MAPK in Lo Vo cells.Results 1. Resveratrol significantly inhibits proliferation, promotes apoptosis and arrests the cell cycle in Lo Vo cells. 2. Resveratrol inhibits the tumor growth in xenograft colon cancer. 3. Resveratrol up-regulates BMP9, but decreases BMP/Smad signaling transduction in Lo Vo cells. 4. Over-expression of BMP9 strengthen the inhibitory effect of resveratrol in Lo Vo cells, and BMP9 knockdown reduce the inhibitory effect of resveratrol. 5. Resveratrol reduces p-Smad1/5/8 protein levels, and increases the level of p-p38 MAPK. In addition, over-expression of BMP9 further enhances the level of p-p38 MAPK, vice versa. 6. Resveratrol increase the level of p-p38 MAPK in Lo Vo cells. And p38 inhibitor promotes the proliferation of Lo Vo cells and reduces the anti-proliferation effect of resveratrol.Conclusion Resveratrol inhibits the proliferation, promotes apoptosis, and arrests cell cycle in Lo Vo cells. These effects may be mediated by activating p38 MAPK through up-regulating BMP9 partly.