关键词: 榄香烯, 凋亡, 热休克蛋白, 抗肿瘤疫苗

Abstract: The possible relationship between heat shock protein (HSP) and the molecular mechanisms of active immunization of elemene-combo tumor cell vaccine (TCV) was tudied by using murine H22 ascitic hepatic carcinoma and L615 leukemia as the experimental models. Stressors, such as elemene or/and heat shock, were applied to study the following items: treated with different concentrations of elemene and different time course of heat shock, transmission electron microscopy were used to observe the morphological influence on tumor cells. Direct immunofluorescent flow cytometry (FCM) was used to detect the apoptotic rate of H22 and L615 tumor cells and influence on the expression of membrane HSP70 and HSP90. The membrane HSP70-peptide complex was separated and purified from H22 and L615 tumor cells induced by different stressors (elemene, MMC) and then a preliminary in vivo active immunoprophylaxis experiment was carried out using those isolated HSP70-peptide complex against H22 on Balb/c mice and L615 on 615 mice. Under transmission electron microscope, the influence of elemene or heat shock on the two kinds of tumor cells is, to some extent, the same or almost the same, which means the apoptosis and necrosis of the cells. The extent of the influence on the two kinds of tumor cells is different, and L615 leukemia was more sensitive and undergoing bigger morphological changes than H22 hepatic carcinoma. It's further proved by FCM that both elemene and heat shock alone can induce the apoptosis of mice H22 hepatic carcinoma or L615 leukemia cells, with the intensifying of this effect by combining use of the two, the apoptosis of cells from mice L615 leukemia relates closely to the concentration of elemene and the time course of heat shock at 42℃. By separating and purifying Balb/c and 615 mice immunized by HSP70-peptide compound of H22 and L615 tumor cell induced by elemene or MMC, the results showed that all mice survived within one month, while none of the mice in control immunized by buffer D survived.

Key words: Elemene, Apoptosis, Heat shock protein, Tummor vaccine