Effect Of Oridonin-mediated Cytokine Changes On Inflammatory Pathways In Human Gastrointestinal Tumor Cells

Objective To observed oridonin effect of proliferation,morphological features and apoptosis in different human gastrointestinal tumor cells,including BxPC-3/PANC-1 human pancreatic cancer cells,Sgc-7901 human gastric cancer cells and Hct 116 human colorectal cancer cells.Comparing the difference of cytokine expression between treatment with and without oridonin in human gastrointestinal tumor cells.Exploring the anti-cancer mechanism of oridonin,which provided the clinical data for treatment on gastrointestinal tumors.Methods 1.BxPC-3/PANC-1,Sgc-7901 and Hct 116 cells were cultivated sterilely and the best concentration of oridonin in inhibiting the proliferation of BxPC-3,Sgc-7901 and Hct 116 cells were obtained by CCK-8 array(for 24h).2.The effects of oridonin on inflammatory factors including IL-2,IL-4,IL-6,IL-10 and TNF-αin PANC-1,Sgc-7901 and Hct 116 cells were measured by Liquidchip System.3.The effects of oridonin on MMP2 and MMP9 in PANC-1,Sgc-7901 and Hct 116 cells were measured by RT-PCR.4.The effects of oridonin on cytokine IL1β and IL-33 in BXPC-3,Sgc-7901 and Hct 116 cells were measured by Western Blot.5.The effects of oridonin on cytokine IL-33 in BXPC-3 and Hct 116 cells were observed by immunofluorescence staining.6.The effects of oridonin on pancreatic cancer in vivo were observed by tumorigenesis assay in nude mice.7.The effects of oridonin on cytokine IL-33 in BXPC-3 cells transplantation tumor were measured by immunohistochemical.Results 1.CCK-8 array showed that oridonin inhibited the proliferation of BxPC-3/PANC-1,Sgc-7901 and Hct 116 cells in a manner as dose-dependent,the best concentration were 75.6μM and 55.8 μM oridonin for 24 hours at 37℃,5%CO2 in BxPC-3/PANC-1,Sgc-7901 and Hct 116 cells respectively.2.Compared with control group,results showed that oridonin could down-regulate the expressions of inflammatory factor IL-6 in PANC-1,Sgc-7901 and Hct 116 cells by Liquidchip System.3.Compared with control group,RT-PCR results showed that oridonin could down-regulate the expressions of MMP2 and MMP9 in BXPC-3,Sgc-7901 and Hct 116 cells.4.The Western Blot showed that oridonin could down-regulate the expressions of IL-33 and IL-1β in BXPC-3,Sgc-7901 and Hct 116 cells compared with the control.5.The immunofluorescence staining showed that oridonin could down-regulate the expressions of IL-33 in BXPC-3 and Hct 116 cells compared with the control.6.The tumorigenesis assay in nude mice showed that oridonin could inhibit the tumor growth.7.The immunohistochemical showed that oridonin could down-regulate the expressions of IL-33 in BXPC-3 cells transplantation tumor.Conclusion 1.Oridonin could inhibit the proliferation of gastrointestinal tumor cells,inhibit the cytoactive of gastrointestinal tumor cells.2.Oridonin could down-regulate the secretion of inflammatory factors IL-33 and IL-1β in BxPC-3/PANC-1,Sgc-7901 and Hct 116 cells.Then the inflammatory factor signal pathway may be influenced by oridonin.3.Oridonin ihibit the gastrointestinal tumor growth in nude mice.