| Objective:The present study was designed to investigate antitumor activities of commonly chemotherapeutic agents combined with Berberine (BBR) against variety cancer cells, to explore the potential mechanisms of two drugs against cancer cells in vivo and in vitro.Methods:1. nine strains of cancer cell lines were selected,which were gastric cancer cell line SGC-7901, the MKN-45, hepatocellular carcinoma cell line SMMC-7901, esophageal cancer cell lines Eca109,colon cancer cell line HT-29, Bile duct cancer cells QBC-939, pancreatic cancer cell line SW1990, breast cancer cell line MCF-7, doxorubicin resistant breast cancer cell line MCF-7/ADR;2. Select some commonly chemotherapeutic drugs which treat the different solid tumor;broth microdilution board method was used to study the synergism of berberine and some chemotherapeutic against the nine strains of tumor cell lines,in order to find the drug that combined with berberine possesses more powerful antitumor activity;3. Through the study on MTT, the growth curve and in vivo experiments,we investigated the mechanism of action of BBR combined with hydroxy camptothecin (HCPT) against gastric cancer cell line SGC-7901, MKN-45;4. Reactive oxygen species (ROS) generation,mitochondrial membrane potential (⊿ψm) and Western blotting were applied to determine the apoptosis of the gastric cancer cells.Results:We investigated the more powerful synergic effect in vitro between berberine and HCPT against gastric cancer cell line SGC-7901, MKN-45by checkerboard microdilution assay and the combination index was0.31±0.19,0.35±0.23, respectively, less than1; MTT were performed to investigate the BBR alone or in combination with HCPT in a dose-dependent against gastric cancer cell line SGC-7901, MKN-45.Morever, After treated with16μg/mL berberine in combination with0.5μg/mL HCPT,combination index was0.38±0.042,0.39±0.06,respectively.The inhibition effect of cell growth of BBR and HCPT using in combination is stronger than that of BBR or HCPT alone respectively (P<0.001). Growth curve further confirmed the synergism of the two drugs against gastric cancer in a time-dependent;10mg/kg BBR (i.p.) in combination with2.5mg/kg HCPT (i.v.) were treated the solid tumor in human gastric cancera model system.The results in vivo showed that the combination of tumor volume and weight were significantly lower than BBR or HCPT respectively (21days). Howerver,10mg/kg BBR (i.p.) in combination with0.5mg/kgHCPT (i.v.) had no therapeutic efficacy.We can investigated the mechanism of acion of BBR combined with HCPT against gastric cancer cell line SGC-7901, MKN-45through apoptosis related proteins analysis, oxidation damage effect analysis;The significant apopsis rates of gastric cancer treated with BBR alone or in combination with HCPT were determined by Flow cytometry instrument detection. We also found that endogenous reactive oxygen species (ROS) was increased, mitochondrial membrane potential and intracellular ATP level were decreased after treating the combination. We investigated the apoptosis related proteins using Western blot method.The result showed that the combination can increase apoptosis. Therefore, the apopsis was induced by the combination of BBR and HCPT through a mitochondria pathway to promote cell DNA damage in human gastric cancer.Conclusions:The results in vivo and in vitro showed the synergism of berberine and hydroxycamptothecine against gastric cancer. |
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