榄香烯增强瘤苗免疫原性的研究

Study on the increased tumor cell immunogenicity induced by elemene

目的 在基因水平上通过与热休克处理对比探讨榄香烯增强肿瘤细胞免疫原性的分子机制。方法 人肝癌细胞HepG2 经 5 0 μg ml榄香烯或 42℃热休克处理后 ,与未经处理的对照组一起 ,分别提取总RNA、逆转录成cDNA、体外转录制备生物素标记的cRNA ,应用含有 12 5 5 0条人类基因的表达谱芯片 ,与热休克对比分析经榄香烯处理的人肝癌细胞HepG2 基因表达谱的改变。结果 榄香烯能使HepG2 细胞的 2 4条基因表达明显改变 ,在这 2 4条基因中有 14条基因在热休克处理时未发生表达变化。结论 榄香烯有可能在基因水平上改变肿瘤细胞的免疫原性 ,并可能诱生与热休克不同的免疫原性物质。

Objective Comparing with heat shock to study the molecular mechanism of increased tumor cell immunogenicity induced by elemene in the level of gene. Methods Total RNA was abstracted from human hepatoma cell line HepG 2 cells untreated or treated with elemene (50μg/ml, 1 h) or heat shock (42℃, 1 h), cDNA was synthesized with reverse transcriptase, cRNA was transcripted from cDNA in vitro successively. The sample and positive control RNA were added into the same Genechip (including 12 550 human genes), hybridized, stained and scanned. The level of gene expression was compared and judged from the intensity of fluorescence with Gene Array Scanner and GENECHIP 3.1 software. Results There were 24 genes with altered expression in HepG 2 cells treated with elemene, 13 of them up-regulated, they were HSP70EP, XRCC4, Rb-protein (mutated), Id-1, NGFR, hEGR1, HTF10, Max, HNF-6, VDUP-1, DAXX, Retina cDNA, KIAA1055. The 11 down-regulated genes were CDC25Hu2, c-fos, bbc-3, metalloproteinase inhibitor, HSF1, HSPA2, Spliceosomal Protein Sap62, KIAA0819, KIAA1053, 24503, 24519. Comparatively, among these genes with elevated or reduced expression observed in HepG 2 cells treated with elemene, 14 of them appeared unchanged in the treatment of heat shock. Conclusion Elemene may change the immunogenicity of tumor cells in the level of gene, and may induce different immunogenic substances from tumor cells treated by heat shock.