The Effects Of Liu Wei Di Huang Bolus On The Express Of Cx And GJIC Of Malignant Melanoma

Malignant melanoma is the malignant tumor which comes from melanin cell. Which is a type of melanin cell malignant tumor which originates neural crest .The outbreak rate is 1% to2% of the whole body malignant tumor. Its malignant degree is high, differing after preparing. With the development of molecular biology , the suicide gene therapy of the tumor has represented the good application prospect as a kind of new approach. But the problem in this remedy is insufficient killing ability, low efficiency of transfection and uncertain safety. It' s reported that increasing the bystander effect is a good approach to resolve this problem. Considering the most important mechanism of bystander effect conducted by the gap junction intercellular communication (GJIC) of cell, researchers today enhance the bystander effect by method of medical inducement to elevate GJIC. But there were not many medicines especially natural medicine with this effect had been found until today. Although some ingredients of TCM had been proved by the clinic or basic investigations on the effects of inducing apoptosis of tumor cells and increasing immunity of the patients, few articles were released about the investigation on increasing bystander effect and cooperating the tumor gene therapy with TCM or their ingredients. At the same time, research in the past proves that: Liu Wei Di Huang Bolus can be able to improve suicide gene bystander. The mechanism of gap junction is other one of bystander effect. Guess that Liu Wei Di Huang Bolus may be improving the GJIC function, which strengthen suicide gene bystander.This research applies the method that the serum pharmacology of the Chinese herbal medicine compound prescribe learns, With malignant melanoma of mou-se cell for research object, We study the function mechanism research of enhancingutility of serum pharmacology of Liu wei Di huang Bolus on "bystander effect"of suicide gene therapy of malignant melanoma. We provid experimentdata, science basis and work foundation to treatment project of suicide genetherapy consociatio-n serum pharmacology of the Chinese herbal medicinecompound prescribe .Objective:During the research, we hoped to explore the possible mechanism of adjusting cell GJIC and inducing of cell apoptosis with Serum Phamacology of LiuweiDiduang Bolus, then we could get the further comprehension about the mechanism of increasing bystander effect with Serum Phamacology of LiuweiDiduang Bolus. Methods:1. Preparation of the herbs serum of LiuweiDiduang Bolus and control serum:SD rat (230～280g), divided into two groups: one was control serum group, the other was Serum Phamacology of Liuwei Diduang Bolus (Administration: 6～8 times of isoeffect dosage). Getting blood from abdominal aorta after the Intragastric administration for one hour, Conservation of serum: -70°C frige.2. The suitable Medication level of the herbs serum of LiuweiDiduang Bolus and control serum. Using MTT assay to detect the sensitivity of herbs serum of Liu Wei Di Huang Bolus and control serum on B16 cell. with the results to decide the suitable medication level.3. The effects of the herbs serum of Liu Wei Di Huang Bolus on GJIC of malignant melanoma. the effects of herbs serum of Liu Wei Di Huang Bolus on the GJIC of B16 cell were detected by SLDT and FACS cell analyzer.4. The effects of the herbs serum of Liu Wei Di Huang Bolus on C_X of B16. the effects of herbs serum of Liu Wei Di Huang Bolus on C_X of B16 were detected by western blot and FITC.Results:1. The preparation of rat serum and the addition of the herbs serum of Liuwei Diduang BolusThe control serum and herbs serum of Liuwei Diduang Bolus showed a promotion effect on B16 cell when the final concentration was 5%～15% but showed inhibiting effect when higher than 20%(micro. Observation: the cells transformed into vacuoles). There was no statistical difference comparing the influences between the herbs serum of Liuwei Diduang Bolus and control serum on CBRH7919. (P>0. 05) 2. The effects of the herbs serum of Liu Wei Di Huang Bolus on GJIC of malignant melanomaThe result of SLDT and FACS cell analyzer showed that the ability of GJIC function of B16 was up-regulated by herbs serum of Liu Wei Di Huang Bolus and control serum. But why control serum can strengthen the ability of GJIC function of B16, the concrete mechanism pending further studies. Along with the density of herbs serum of Liu Wei Di Huang Bolus increase, its GJIC function obtains the enhancement, This indicated the ability of GJIC function of B16 was up-regulated by herbs serum of Liuwei Diduang Bolus.3. The effects of the herbs serum of Liu Wei Di Huang Bolus on C_X protein of B16.The result of western blot and FITC showed that C_X was up-regulated by herbs serum of Liu Wei Di Huang Bolus and control serum. But why control serum can strengthen C_X, the concrete mechanism pending further studies. Along with the density of herbs serum of Liuwei Diduang Bolus increase, The connexion expression has the enhancement, This indicated the expression of C_X was up-regulated by herbs serum of Liu Wei Di Huang Bolus.Conclusion:This research discussed The Mechanism Research of herbs serum of LiuWei Di Huang Bolus on GJIC of malig-nant melanoma .The analysis result indicated Liu Wei Di Huang Bolus strengthens the express of C_X of malignant melanoma, Which can strengthen the function of gap junction. The mechanism of gap junction of bystander effect may be one of the Liu Wei Di Huang Bolus mechanism to suicide gene treatment synergy.