三氧化二砷诱导人肺腺癌细胞凋亡形态学及数量变化

摘要

Objective To observe the cell proliferation, morphology change and count number change of the AS49 cells under the arsenic trioxide (As2O3) induction. Methods The AS49 cells were cultured into logarithmic growth phase in vitro, and the cells were divided into experimental group and blank group. As2O, was added into experimental group, the apoptosis status of the cells was observed under light microscope and by electron microscope scanning. The flow cytometer was used to count the cell number and calculate the change of the apoptosis ratio. Results Under the light microscope, the experimental group cells were deflated, and turned into a circle, the transmissivity decreased, and the vacuolus changes were detected in the cell body. After fluorescent staining by the acridine orange (AO) , the fluorescence intensity and density of the attached cells decreased , part of the cells broke apart, nuclei disappeared, and the cell apoptosis appeared. Under the electron microscope we saw karyopyknosis, karyorrhexis, lose nuclear membrane, chromatin margination, and also the apoptotic body. The count number indicated that the apoptotic ratio (14.497% )of the experimental group was higher than that (0. 027% )of the blank group (χ2 =18.447, P<0.0001). Conclusion Inducted by As2O3, the proliferated and apoptic changes appeare conclusively in A549 cells with an increase in the number of apoptic cells, suggesting that As2O3 might be a new option in treatment of lung adenocarcinoma.%目的 观察三氧化二砷( arsenic trioxide,As2O3)诱导人肺腺癌A549细胞增殖和凋亡的形态学改变及计数变化.方法 体外培养A549细胞至对数生长期后随机分为实验组和空白对照组.实验组加入As2 O3药液,光学显微镜(光镜)和电子显微镜(电镜)扫描观察细胞凋亡情况,使用流式细胞仪定量计数凋亡细胞比例变化,并与空白对照组比较.结果 实验组A549细胞光镜下表现为细胞缩小、变圆,透光度降低,胞体呈空泡样改变；经吖啶橙荧光染色后,贴壁细胞荧光强度降低,密度下降,部分细胞破碎、胞核消失,出现凋亡细胞；电镜下可见细胞核固缩、破裂,无核膜包被,核染色质边集胞膜,出现凋亡小体.计数结果表明,实验组凋亡细胞比例( 14.497％)明显多于空白对照组(0.027％),差异有统计学意义(x2 =18.447,P＜0.0001).结论 经As2O3诱导后,A549细胞发生了明确的增殖和凋亡改变,且凋亡细胞计数增加,故As2O3有可能成为临床治疗肺癌的一种新方法.