目的探讨β-榄香烯对肾癌GRC-1细胞体外放射增敏相关基因表达谱的影响。方法体外培养GRC-1细胞,MTT法检测不同放射剂量(0、50、1。O、150及200cGy)下,不同浓度β-榄香烯(空白组、空白乳组及10、20、30、40、50、60、70、80以及90μg/ml β-榄香烯组)对GRC-1细胞体外生长的影响。流式细胞仪检测单纯放疗组(空白组,150cGy)和放疗增敏组(20μg/ml β-榄香烯,150cGy)细胞周期变化与凋亡。选取包含4096个cDNA基因表达谱芯片分析2组间基因谱表达差异。结果β-榄香烯在20μg/ml时对GRC-1细胞有体外放射增敏作用。对肾癌细胞G2M阻滞作用随时间增加而增强,24h时凋亡率为17.26%,48h时作用达最高峰,凋亡率为24.34%。随时间和照射剂量增加细胞凋亡水平增高。基因芯片筛选出2组间差异表达基因360条,上调基因265条、下调基因95条。结论β-榄香烯乳对肾癌细胞的放疗增敏作用可能涉及原癌、抑癌基因等多种相关基因的变化,是一个多基因参与的复杂事件。
Objective cDNA microarray technology was applied for screening genes associated with increasing radiate sensitivity by β-elemenen. Methods GRC 1 cells were cultured in vitro. The effect of increasing radiate sensitivity of β-elemenen was measured by using MTT methods,in order to decide the doses of the β-elemenen and radiation in the radiate sensitivity group. Flow cytometry(FCM) was used to measure cell cycle and apoptosis in radiotherapy group and increasing radiate sensitivity group. The biostar H-40s gene chip was applied to investigate the differentially expressed genes. Results MTT showed β-elemenen could enhance the radiate sensitivity to GRC-1 cells in vitro. FCM showed that the G2 M stage blocking caused by 20 μg/ml β-elemenen was enhanced with time increase, it reached peak at 24 hours. The level of apoptosis enhanced with increase of drug-dose and time. A total of 360 genes were expressed differently between two groups. Conclusions The effects of increasing radiate sensitivity caused by β-elemenen may be associated with proto-oncogene and anti-oncongene. The protein of cell cycles the associated protein of cytoskelet on and cell movement, the protein of DNA synthesis, reparation and recombination, DNA integrate, transcribe and transcription factor, the receptor,signal of cell and transfer protein, cell metabolism growth, immunization and protein interpretation may also be involved. It could be a complex event with multi-genes participating.