BACKGROUND: Midkine is a heparin-binding growth factor that promotes the proliferation, survival, migration and differentiation of various target cells. Midkine plays an important role in tumorigenesis and tumor progr...BACKGROUND: Midkine is a heparin-binding growth factor that promotes the proliferation, survival, migration and differentiation of various target cells. Midkine plays an important role in tumorigenesis and tumor progression, and is overexpressed in many human malignant tumors. Patients with high tumor midkine expression frequently have a worse prognosis than those with low expression. The present study was designed to investigate the interaction network of midkine in hepatic cancer cells, and to elucidate its role in hepatocellular carcinoma. METHODS: DNA encoding full-length midkine was cloned into pDBLeu vector to serve as bait in yeast two-hybrid screening to identify interacting proteins. Candidate proteins were examined on SC-Leu-Trp-His+3-AT (20 mmol/L) plates and assayed for X-gal activity, then sequenced and classified according to the GenBank. Finally, identified proteins were expressed by the in vitro expression system pCMVTnT, and protein interactions were confirmed by co-immunoprecipitation. RESULTS: Using the yeast two-hybrid system, we found 6 proteins that interacted with midkine: NK-kappa-B inhibitor alpha (I-κ-B-α), Dvl-binding protein naked cuticle 2, granulin, latent active TGF-β binding protein 3, latent active TGF-β binding protein 4, and phospholipid scramblase 1. In vitro co-immunoprecipitation demonstrated that all identified proteins directly interacted with midkine.CONCLUSION: The identification of midkine-interacting proteins in hepatic cancer cells indicates that midkine is a multifunctional factor that may participate in cell migration, differentiation, and proliferation, and is also associated with the multicellular response feedback during the development of hepatocellular carcinoma.展开更多
Background Recent studies have shown that effective ingredients of Chinese herbs are used more and more widely in the treatment or co-treatment of cancers, however, they are usually used separately and there has been ...Background Recent studies have shown that effective ingredients of Chinese herbs are used more and more widely in the treatment or co-treatment of cancers, however, they are usually used separately and there has been limited research about joint application of Chinese herbs in multi-modal treatment. The aim of this study was to screen a QHF （Q： Qingrejiedu, H： Huoxuehuayu and F： Fuzhengguben） formula for effective ingredients from Chinese medicines and assess its anti-hepatic cell cancer （HCC） effect in combination with chemotherapy.Methods Six effective ingredients from Chinese medicine were selected based on the previous literature and used in the study. The QHF formula and the best ratio of ingredients were evaluated in H22 mouse （KM） models with solid tumors and ascites tumors by uniform design and monitoring inhibition of tumor growth and survival. We then observed the anti-hepatic cell cancer （HCC） effect of QHF when combined with cisplatin （DDP） in H22 mouse （Balb/c） models with solid tumors and ascites tumors. Evaluating of the therapeutic effect included the general condition of the mice, inhibition of tumor growth, survival, changes in body weight, thymus index, spleen index and WBC counts.Results The optimal QHF dose ratio for anti-hepatic cell cancer treatment was： 800 mg/kg Cinobufotalin, 14 mg/kg Ginsenosides Rg3, 5.5 mg/kg PNS and 100 mg/kg Lentinan. Treatment was more efficient in inhibiting the growth of transplanted tumors in H22 mice when using the QHF formula （55.91%） than using Cinobufotalin （33.25%）, Ginsenosides Rg3 （35.11%）, PNS （27.12%） or Lentinan （4.97%） separately. QHF also prolonged the life of H22 ascites hepatic cancer mice more efficiently （38.13%） than Cinobufotalin （25.00%）, Ginsenosides Rg3 （27.27%）, PNS （23.30%） or Lentinan （24.43%）. QHF combined with DDP could reduce DDP-induced leucopenia, spleen and thymus atrophy and other toxic reactions. Combining QHF with DDP the tumor growth inhibition reached 82.54% with a 66.83% increase in survival. Conclusions QHF is more efficient in anti-hepatic cell caner treatment than the single drugs that constitute the formula. QHF combined with DDP can attenuate tumor growth and suppresses the DDP-induced toxic reactions.展开更多
基金supported by grants from the National Natural Science Foundation of China (30772534)the Natural Science Foundation of Huzhou (2010YS05)
文摘BACKGROUND: Midkine is a heparin-binding growth factor that promotes the proliferation, survival, migration and differentiation of various target cells. Midkine plays an important role in tumorigenesis and tumor progression, and is overexpressed in many human malignant tumors. Patients with high tumor midkine expression frequently have a worse prognosis than those with low expression. The present study was designed to investigate the interaction network of midkine in hepatic cancer cells, and to elucidate its role in hepatocellular carcinoma. METHODS: DNA encoding full-length midkine was cloned into pDBLeu vector to serve as bait in yeast two-hybrid screening to identify interacting proteins. Candidate proteins were examined on SC-Leu-Trp-His+3-AT (20 mmol/L) plates and assayed for X-gal activity, then sequenced and classified according to the GenBank. Finally, identified proteins were expressed by the in vitro expression system pCMVTnT, and protein interactions were confirmed by co-immunoprecipitation. RESULTS: Using the yeast two-hybrid system, we found 6 proteins that interacted with midkine: NK-kappa-B inhibitor alpha (I-κ-B-α), Dvl-binding protein naked cuticle 2, granulin, latent active TGF-β binding protein 3, latent active TGF-β binding protein 4, and phospholipid scramblase 1. In vitro co-immunoprecipitation demonstrated that all identified proteins directly interacted with midkine.CONCLUSION: The identification of midkine-interacting proteins in hepatic cancer cells indicates that midkine is a multifunctional factor that may participate in cell migration, differentiation, and proliferation, and is also associated with the multicellular response feedback during the development of hepatocellular carcinoma.
文摘Background Recent studies have shown that effective ingredients of Chinese herbs are used more and more widely in the treatment or co-treatment of cancers, however, they are usually used separately and there has been limited research about joint application of Chinese herbs in multi-modal treatment. The aim of this study was to screen a QHF （Q： Qingrejiedu, H： Huoxuehuayu and F： Fuzhengguben） formula for effective ingredients from Chinese medicines and assess its anti-hepatic cell cancer （HCC） effect in combination with chemotherapy.Methods Six effective ingredients from Chinese medicine were selected based on the previous literature and used in the study. The QHF formula and the best ratio of ingredients were evaluated in H22 mouse （KM） models with solid tumors and ascites tumors by uniform design and monitoring inhibition of tumor growth and survival. We then observed the anti-hepatic cell cancer （HCC） effect of QHF when combined with cisplatin （DDP） in H22 mouse （Balb/c） models with solid tumors and ascites tumors. Evaluating of the therapeutic effect included the general condition of the mice, inhibition of tumor growth, survival, changes in body weight, thymus index, spleen index and WBC counts.Results The optimal QHF dose ratio for anti-hepatic cell cancer treatment was： 800 mg/kg Cinobufotalin, 14 mg/kg Ginsenosides Rg3, 5.5 mg/kg PNS and 100 mg/kg Lentinan. Treatment was more efficient in inhibiting the growth of transplanted tumors in H22 mice when using the QHF formula （55.91%） than using Cinobufotalin （33.25%）, Ginsenosides Rg3 （35.11%）, PNS （27.12%） or Lentinan （4.97%） separately. QHF also prolonged the life of H22 ascites hepatic cancer mice more efficiently （38.13%） than Cinobufotalin （25.00%）, Ginsenosides Rg3 （27.27%）, PNS （23.30%） or Lentinan （24.43%）. QHF combined with DDP could reduce DDP-induced leucopenia, spleen and thymus atrophy and other toxic reactions. Combining QHF with DDP the tumor growth inhibition reached 82.54% with a 66.83% increase in survival. Conclusions QHF is more efficient in anti-hepatic cell caner treatment than the single drugs that constitute the formula. QHF combined with DDP can attenuate tumor growth and suppresses the DDP-induced toxic reactions.
