The effect of calmodulin antagonist berbaminederivative-EBB on hepatoma in vitro and in vivo / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To evaluate the anti-hepatoma effect of Calmodulin antagonist 0 - 4-ethoxyl-butyl-Berbamine (EBB), one of the berbamine derivatives.</p><p><b>METHODS</b>Monotetrazolium (MTT) method was used to analyze the effect of EBB on the proliferation and growth inhibition effect. Of a hepatoma cell line in vitro. A mouse hepatoma model was induced by injection of hepatoma cells (H22) in the abdominal cavity. The effect of EBB on survival at different concentrations as well as in combination with 5-FU were investigated in vivo. Flow cytometry analysis, dot blot hybridization, western blot, immunochemistry, enzyme-linked lectin assay (ELISA), trifluoperazine (TFP) and electron microscopic observation were used to study the effect of EBB on cell cycle process, P53 mRNA and protein levels, calmodulin content and ultrastractural changes of hepatoma cells.</p><p><b>RESULTS</b>EBB exerts a very strong inhibitory effect on human hepatoma cell line 7402 and mouse hepatoma cell line H22 in vitro. The IC(50) value of EBB for the two cell lines are 3.312 microg/ml and 1.167 microg/ml, respectively. The sensitivity of H22 cells to 5-FU can be markedly enhanced: The IC(50) dosage of 5-Fu can be decreased from 0.75 microg/ml down to 0.15 microg/ml, when jointly administered with nontoxic dosages of EBB (IC(10)). In vivo, EBB can prolong the lifespan of mice with ascites H22 to more than three months. 64% of mice survived, while all animals in the control group died by the 18th day. When EBB (5 mg x kg(-1) x d(-1)) is jointly used with 5-FU (25 mg x ml(-1) x d(-1)), 73% of mice with ascites H22 survived, much higher than 27% in the 5-FU treated group. EBB can enhance the anti-hepatoma ability of 5-Fu treatment. EBB mechanism against hepatoma: P53 expression in the EBB treated group is substantially higher than that in the control group. EBB increased the translation of P53. As a calmodulin antagonist, EBB decreases amount of the CaM in hepatoma cells and blocked the hepatoma cell proliferation cycle at the G(2)M phase. Before the G(0)/G(1) phase, a diploid peak and apoptic cells in the treated groups were observed.</p><p><b>CONCLUSIONS</b>The CaM antagonist, EBB, has a strong anti-hepatoma effect and enhances the effect of 5-FU, induces hepatoma cell to apoptosis, promotes the P53 protein expression and decreases the amount of CaM in the cytoplasm. All these results demonstrate that EBB is a new and potentially useful drug against hepatoma and should be researched further.</p>

Giant Tumor Cells in the Bone Marrow and Their Significance / 中国实验血液学杂志

Giant tumor cells and their varieties in the bone marrow were found in 7 patients with abnomal hematopoiesis phenomena. These cells were artificially devided into 5 kinds according to the difference of their morphology. Most of these cells were corresponding to lymphoid-monocytoid-macrophagocytoid cells with Wright's staining, cytochemical stainings, immunocytochemical stainings, flow cytometry examination, electron microscopy and pathologic study. The bone marrows were hypercellular and marked dysplastic hematopoiesis phenomena. Two of the 7 cases were diagnosed as malignant lymphoma with bone marrow biopsy. All cases characteristically showed no lymph node enlargement or hepatosplenomegaly or any local tumor mass. As to the prognosis of these cases, two patients died with survival time of 8 and 17 months, respectively, one was on critical condition at course of 10 months, and the other 4 cases were in comparatively stable condition with courses of 2.5 to 24 months. These patients seem to be a group of rare malignant lymphoid-monocytoid-macrophagocytoid proliferative diseases.