冬青素诱导宫颈癌HeLa细胞凋亡及其机制初步研究

Study on apoptosis of human cervical cancer HeLa cell induced by Ilexgenin A and its primary mechanism

目的:研究冬青素(IA)对人宫颈癌HeLa细胞凋亡的诱导作用及其相关机制。方法:建立人宫颈癌HeLa细胞体外培养体系,采用CCK-8法检测细胞活力,MTT法测定乳酸脱氢酶(LDH),免疫比色法检测细胞增殖情况,DAPI荧光染色观察细胞凋亡形态学变化,Western Blot检测Caspase-3/7和NF-κB蛋白表达水平。结果:用冬青素处理过的HeLa细胞,LDH释放增加,呈剂量和时间依赖性。在CCK-8和5-溴脱氧尿嘧啶核苷(BrdU)测定过程中,冬青素比标准药物抗HeLa活性更强,随浓度和作用时间的增加,细胞的活性不断下降,对HeLa增殖的抑制作用明显。DAPI染色观察用冬青素处理过的HeLa细胞浓缩明显,细胞核分散,染色质凝聚。Western Blot结果显示用冬青素处理过的HeLa细胞Caspase-3/7的活性显著增加(P<0.01),NF-κB蛋白表达水平下降(P<0.01)。结论:冬青素具有诱导HeLa细胞凋亡的作用,其机制可能与激活Caspase-3/7和抑制NF-κB蛋白表达有关。

Abstract： Objective： To investigate the apoptosis of human cervical cancer HeLa cell induced by llexgenin A （IA） and its primary mechanism. Methods： The viability of HeLa cells was detected by the method CCK-8. The activity of LDH was measured by MTT assay. Immune colorimetry was used to test the proliferation of Hela cells. The change of morphology of apoptosis HeLa cells was observed by DAPI fluorescence staining. Western Blot was used to detect the expressions of Caspase-3/7 and NF-xB proteins of HeLa cells. Results： After HeLa cells were treated by IA, the leakage of LDH increased in a dosage and time dependent manner. In the process of determination of BrdU and CCK-8, the inhibitory effect of IA to HeLa cell was stronger than standard drugs, and showed clear inhibitory effect on the HeLa cell proliferation with the increasing of concentration and the acting time. By using the DAPI fluorescence dyes, the HeLa cell cytoplasm concentrating, nucleolus dispersion and chromatin aglomeration were observed. Western Blot analysis showed that the expression of Caspase-3/7 was significantly enhanced and the expression of NF-κB proteins was down-regulated. Conclusion： IA could induce the apoptosis of HeLa cells and its mechanism is related to activate the activity of Caspase-3/7 and down-regulate the expression of NF-κB protein.