MAP30-PEG结合物对A549体外细胞增殖作用的研究

目的探讨苦瓜籽核糖体失活蛋白MAP30及其聚乙二醇修饰物对人肺腺癌细胞A549增殖、细胞凋亡的影响和caspase3相关凋亡途径的诱导作用。方法用MTT法测定MAP30和MAP30-PEG结合物对人肺腺癌细胞A549的抑制率;通过Hoechst33258荧光染色观察A549细胞凋亡形态,并用流式细胞术测定细胞凋亡率以及用western blot检测凋亡细胞caspase3表达。结果 MAP30和MAP30-PEG结合物均对A549有抑制作用,且呈时间和剂量依赖性关系;MAP30-PEG结合物对A549细胞增值抑制率约为MAP30的60%;Hoechst33258荧光染色后均呈典型凋亡形态;流式细胞术检测显示,MAP30和MAP30-PEG结合物作用于A549细胞48 h后均可使A549细胞呈现Sub-G1凋亡峰,凋亡率分别为58.7%、42.7%;MAP30和MAP30-PEG结合物对胞内caspase3在凋亡过程中显示有激活作用。结论 MAP30和MAP30-PEG结合物均对A549增殖有明显的抑制作用;两者均能通过caspase3途径诱导细胞凋亡。

Effects of MAP30-PEG conjugates on the proliferation of A549 cells in vitro

OBJECTIVE To study the effect of MAP30 and MAP30-PEG conjugates on the proliferation,apoptosis and inducing effect on caspase3 pathway in lung cancer A549 cells.METHODS MTT assay was used to investigate the effect of MAP30 and MAP30-PEG conjugates on proliferation of A549 cells.After the treatment with MAP30 and MAP30-PEG conjugates,morphological changes of A549 cells and apoptosis proportion were determined by Hoechst33258 fluorescent staining and flow cytometry(FCM),respectively.Protein expression of caspase3 was detected by Western blot.RESULTS Both native and PEGlyated MAP30 significantly inhibited the growth of lung cancer A549 cells in a time-and dose-dependent manner.MAP30-PEG conjugate preserved about 60% antiproliferative activity of native averagely.After staining with Hoechst33258,typical morphologic changes of apoptosis cells were observed in both groups treated with the drugs.FCM showed that a typical sub-diploid apoptosis peak was demonstrated in both groups treated with MAP30 and MAP30-PEG conjugates for 48 h.Native and PEGlyated MAP30 induced a different apoptotic rate,which was 58.7% and 42.7%,respectively.The expression of caspase3 significantly increased in both groups treated with the drugs at 48 h.CONCLUSION MAP30 and MAP30-PEG conjugates inhibited the proliferation of A549 cells significantly in vitro.Both of two drugs can induce the apoptosis of A549 cells and its mechanism may be through caspase3 pathway.