家蝇幼虫血淋巴蛋白MAC-1诱导人宫颈癌细胞凋亡的实验
Apoptosis of Cervical Cancer Cell Line HeLa Induced by MAC-1 Protein from Musca Domestica Larva Hemolymph
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摘要: 目的 探讨家蝇幼虫血淋巴蛋白(anticancer protein from Musca domestica larva hemolymph，MAC-1)诱导人宫颈癌细胞系（Cervical cancer cell lines，HeLa）的凋亡过程中fas、caspase 8、bcl-2凋亡调节基因的相互关系及可能的作用机制。方法苏木素-伊红（HE）染色法、流式细胞术（FCM）观察MAC-1蛋白作用细胞凋亡及增殖变化，应用免疫细胞化学技术检测MAC-1蛋白作用前后凋亡相关基因fas、caspase 8和bcl-2表达的变化。结果MAC-1蛋白诱导后HeLa 细胞形态发生改变；流式细胞仪分析结果发现细胞凋亡率随蛋白作用时间的延长和蛋白浓度的增加而降低的趋势。免疫细胞化学法提示MAC-1蛋白作用HeLa细胞后fas 和caspase 8的染色增强（P<0.05），bcl-2的染色减弱（P<0.05）。结论家蝇幼虫血淋巴蛋白MAC-1能够抑制HeLa 细胞增殖并诱导凋亡，此作用随着蛋白的浓度增加和作用时间延长反而降低，其机制可能与fas、caspase 8和bcl-2表达有关。Abstract: Objective To investigate the apoptosis-inducing effect and mechanism of the MAC-1 protein from Musca domestica larvae three age hemolymph extract on Cervical cancer cell line HeLa. Methods The morphology of apoptosis cells was observed with hematoxylin- eosin stainings and flow cytometry.The expression of Fas，caspase 8 and bcl-2 were examined by immunocytochemical staining Results. Results Treated with the MAC-1 protein from Musca domestica larvae three age hemolymph extract，the HeLa cells showed the morphological apoptosis;FITC/PI stained FCM showed the apoptosis rate of HeLa had been decreased along with the prolonging of time and the increase of the MAC-1 protein concentrations.With the MAC-1 protein，immunocytochermical staining showed the enhanced expression of Fas and caspase 8，and attenuated expression of bcl-2. Conclusion The MAC-1 protein from Musca domestica larvae three age hemolymph can induce HeLa cells，tapoptosis in- time and concentration -dependent manners，which may be related to the expression of Fas，caspase8 and bcl-2.
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