目的 研究冬凌草甲素（ORI）對人源結(jié)腸癌細胞HCT116的增殖抑制作用，并探討其可能的分子機制。方法 采用結(jié)晶紫染色研究ORI （5、10、15、20、25 μmol/L）對HCT116細胞的增殖抑制作用；采用Western blotting技術(shù)研究ORI （10、15、20 μmol/L）對HCT116細胞中增殖細胞核抗原（PCNA）、凋亡相關(guān)指標（Caspase-3和Cleaved-caspase-3）、p38 MAPK信號通路相關(guān)指標（Smad1/5/8、p-Smad1/5/8、p38、p-p38）蛋白表達的影響；采用Western blotting、凝膠電泳技術(shù)分析ORI對BMP7蛋白、mRNA表達的影響；轉(zhuǎn)染重組BMP7腺病毒（AdBMP7）實現(xiàn)BMP7的外源性過表達，應(yīng)用BMP7抗體（anti-BMP7）降低BMP7水平，檢測ORI是否通過調(diào)節(jié)BMP7表達影響HCT116細胞增殖凋亡、p38 MAPK信號通路。結(jié)果 與對照組比較，ORI明顯抑制HCT116細胞增殖，下調(diào)PCNA蛋白水平；明顯上調(diào)Caspase-3、Cleaved caspase-3、p-p38蛋白表達，且呈濃度與時間相關(guān)性，對Smad1/5/8、P-Smad1/5/8蛋白表達無明顯影響；明顯上調(diào)BMP7蛋白及RNA表達；外源性過表達BMP7加強了ORI對上述蛋白表達的調(diào)控，anti-BMP7則部分抑制了ORI的作用。結(jié)論 ORI抑制HCT116細胞的增殖，其機制可能與上調(diào)BMP7蛋白表達進而激活p38 MAPK信號通路相關(guān)。

Objective To investigate the proliferation inhibition effect and the possible mechanism of Oridonin(ORI) on human colon cancer HCT116 cells.Methods The inhibitory effects of ORI (5, 10, 15, 20, 25 μmol/L) on the proliferation of HCT116 cells were tested by crystal violet staining. The effects of ORI (15, 20, 25 μmol/L) on the expression of proliferating cell nuclear antigen (PCNA), apoptosis related index (Caspase-3 and Cleaved-caspase-3) and p38 MAPK signal pathway related protein (Smad1/5/8, p-Smad1/5/8, p38 and p-p38) in HCT116 cells were studied by Western blotting assay. The effect of ORI on the expression of BMP7 protein and mRNA was analyzed by Western blotting and gel electrophoresis. Next, the exogenous overexpression of BMP7 were realized by transfecting recombinant adenovirus (AdBMP7), and the BMP7 antibody (anti-BMP7), which could reduce the expression of BMP7, was used to detect the influence of ORI on proliferation and apoptosis of HCT116 cells and p38 MAPK signal pathway by regulating the expression of BMP7.Results Compared with control group, ORI could inhibit the proliferation of HCT116 cells obviously, down-regulated the PCNA protein level; Western blotting results showed that ORI up-regulated the protein level of BMP7, Caspaes-3, Cleaved caspase-3 and p-p38 in a time and concentration dependent manner, with no obvious effect on the expression of Smad1/5/8 and P-Smad1/5/8 protein and obvious up-regulative effect on the BMP7 protein and mRNA. Over expression of BMP7 strengthened the regulative level of above-mentioned proteins, while anti-BMP7 partly inhibited such effects of ORI.Conclusion ORI can inhibit the proliferation of HCT116 cells, which may be related to the up-regulation of BMP7 protein expression with view to activating of the p38 MAPK signal pathway.

重慶市渝中區(qū)科委課題（No 20150120）