The Experimental Study Of Sophorcarpidine And Gemcitabine With Thermotherapy Inhibiting Angiogenesis

[Background] The tumor growth divides into the blood vessel time and the bloodvessel earlier period. Tumor cells gain nutrition mainly through dispersion processduring blood vessel earlier period. When the solid tumor grows up to 1～2mm~3 and thetumor cells amount to about 10~7, it has to depend on the newbom blood vessel toprovide enough oxygen and nutrients to maintain further growth. After vascularizationof tumor internal tissue, its volume increases rapidly. The tumor metastasis can beinfluenced by many kinds of factors. The blood route metastasis is the important wayfor distant dispersion of tumors, therefore, tumor angiogenesis plays a vital role intumor metastasis. Considering the importance of tumor angiogenesis to tumor growthand metastasis, to block angiogenesis is a feasible method to treat tumor. At present,high-time, low-dose chemotherapy is the commonly used administration model inclinical activities, whereas successive or high-frequency low dose anti-angiogenicchemotherapy substituted for traditional chemotherapy of routine dose and periodicaldelivery can enhance the curative effect to tumors with drug resistance and decreasesthe toxicity. Moreover, in the tumor tissue, the newborn blood capillary endothelialcells belong to normal cell category, the genetics of which are stable and drugresistance seldom happens to them.Hyperthermia is one method to treat tumor through elevating the temperature oftumor tissue to 40～44℃by heating,namely using related physical energy precipitationin tissue to generate heat effect to raise the temperature of tumor tissue to an effectivetreatment point and maintain it for a period of time, during which tumor cells will bekilled but normal cell uninjured.The anti-angiogenic therapy is to mollify patient’s condition and make them tosurvive with a tumor through stabilizing disease and postponing tumor diffusion bytreatment. This is congruent with the therapeutic principle of strengthening bodyresistance and suppressing pathogenic factors in traditional Chinese medicine. Thepurpose of strengthening body resistance and suppressing pathogenic factors is toprolong survival duration and improve life quality, making the patients to co-existwith tumor peacefully. Researches have shown that many kinds of traditional Chinese medicines and their preparation have the function of anti-angiogenesis, such asginsenoside Rg3, red pigment (an active monomer ingredient withdrawn fromTripterygium wilfordii), hydroxycamptothecin and paclitaxe.[Objectives] This experiment aims to①study antiangiogenetic ability ofSophorcarpidine, Gemcitabine combined with thermotherapy;②observe influance onproliferation of blood vein endothelial cells and growth of chick embryochoriallantoic membrane vessels when using combination of Sophorcarpidine,Gemcitabine.③study the influence of Sophorcarpidine with Gemcitabine onHUVEC migration was evaluated by transwell.[Methods] We use MTT method to observe the influence of Sophorcarpidine,Gemcitabine on human umbilical vein endothelial cells (HUVEC) or LOVOproliferation. The influence of Sophorcarpidine, Gemcitabine on HUVEC migrationwas evaluated by transwell. Chick embryo chorioallantoic membrane (CAM) modelwas used to check whether the neovascularization of CAM could be suppressed invivo.[Results]1. The survival rate of HUVEC is among86.4%-56.1% within the dose of6.25-200μg/ml sophorcarpidine and is negative correlated with the concentration(correlation coefficient is -0.955). The survival rate of LOVO within this doses ismore than HUVEC.2. The survival rate of HUVEC is among79.1%-43.8%within the dose of0.625-20μg/ml gemcitabine and is negative correlated with the concentration(correlation coefficient is -0.881). The survival rate of LOVO within this doses ismore than HUVEC.3. There is a synergedtic effection in anti-angiogenic when combined usesophorcarpidine (50μg/ml,100μg/ml,and 200μg/ml) with thermotherapy in vitro whileaddition effection whin sophorcarpidine (6.25μg/ml,12.5μg/ml,and 25μg/ml) withthermotherapy in vitro. There is a synergedtic effection in anti-angiogenic whencombined use gemcitabine (0.625μg/ml, 1.5μg/ml,5μg/ml and 10μg/ml) withthermotherapy in vitro while addition effection whin gemcitabine (2.5μg/ml,20μg/ml)with thermotherapy in vitro.4. There is a synergedtic effection in anti-angiogenic when combined usesophorcarpidine (25μg/ml, 50μg/ml) with gemcitabine(1.25μg/ml, 2.5μg/ml) in vitro.5. There is a synergedtic effection in inhibited migration of HUVEC in vitro when combined use sophorcarpidine (12.5/μg/ml) with gemcitabine(1.25μg/ml) in vitrowhile addition effection whin sophorcarpidine (6.25μg/ml) with gemcitabine(0.625μg/ml, 1.25μg/ml) in vitro.And there is a synergedtic effection in inhibitedmigration of HUVEC in vitro when combined use sophorcarpidine (12.5μg/ml) withgemcitabine (0.625μg/ml) in vitro.6. Sophorcarpidine can suppress significantly CAM vessels at doses of 12.5-25μg/ml;gemcitabine can suppress significantly CAM vessels at doses of 1.25-2.5μg/ml.[Conclusion] Sophorcarpidine and Gemcitabine can inhibit the proliferation ofhuman microvescular endothelial cells and the formation of chick embryochoriallantoic membrane blood vessels at the doses which do not demonstratesuppressing effect on cancerous cells. This experiment shows that Sophorcarpidineand Gemcitabine have antiangiogenetic ability both in vivo and in vitro. Thecombined use of drugs and thermotherapy has synergistic antiangiogenetic action invitro.