蒲葵子甲醇提取物抗鼻咽癌的实验研究

Experimental study on livistona chinensis methanol extracts against nasopharyngeal carcinoma

目的研究蒲葵子甲醇提取物(LCME)对鼻咽癌细胞的生长抑制作用。方法制备LCME的PBS溶液;CCK8法检测鼻咽癌细胞株C666和5-8F中LCME的半抑制浓度(IC50)值;光学显微镜下观察LCME作用下C666和5-8F细胞的凋亡形态变化;免疫荧光共聚焦检测LCME作用下Caspase3在C666和5-8F细胞中的表达情况;平板克隆实验观察LCME对C666和5-8F细胞克隆形成率的影响;CCK8实验检测LCME对C666和5-8F细胞增生能力的影响;Western Blotting实验检测LCME作用下C666和5-8F细胞中Caspase3,Caspase7和P21表达变化情况;流式细胞术检测LCME作用下C666和5-8F细胞的凋亡率。结果实验结果显示LCME对体外培养的鼻咽癌细胞株C666和5-8F的增生和克隆形成具有明显的抑制作用,并可诱导其发生凋亡。C666的48 h IC_(50)=130μg/L,5-8F的48 h IC_(50)=220μg/L。LCME能诱导鼻咽癌细胞凋亡,使鼻咽癌细胞Caspase3、Caspase7和P21的表达上调。结论 LCME对体外培养的鼻咽癌细胞株C666和5-8F的增生具有明显的抑制作用,并可诱导其发生凋亡,具有开发成为抗鼻咽癌的中药有效制剂的潜能。

Objective To study the effect of Livistona chinensis methanol extracts（LCME） on inhibiting the growth of nasopharyngeal carcinoma cells.Methods LCME in PBS solution was prepared and CCK8 assay was used to test nasopharyngeal carcinoma cell C666 and semi-inhibitive concentration value（IC50） of LCME in 5-8 F cell lines.The morphological characteristics of apoptosis of C666 and 5-8 F cell lines were observed by optical microscope and the expressions of Caspase3 in C666 and 5-8 F cell lines were detected by confocal immunofluorescence.Plate clonality assay was used to measure the effects of LCME on C666 and 5-8 F＇s colony formation rates.CCK8 assay was used to analyze the effects of LCME on the proliferation capacity of C666 and 5-8 F cell lines.Western Blotting was used to detect the changes in Caspase3,Caspase7 and P21 expressions in the C666 and 5-8 F cell lines.Flow cytometry was used to measure the apoptotic rates of C666 and 5-8 F cell lines.Results LCME significantly inhibited the in vitro proliferations and colony formation rates of C666 and 5-8 F cell lines and induces their apoptosis.The IC50 of C666 was 130 μg/L within 48 and the IC50 of 5-8 F was 220 μg/L within 48 h.LCME could induce apoptosis of nasopharyngeal carcinoma cells up regulate expressions of Caspase3,Caspase7 and P21.Conclusion LCME significantly inhibited the in vitro proliferations of C666 and 5-8 F cell lines and induced their apoptosis which makes it potential to be developed as an effective traditional Chinese medicine against nasopharyngeal carcinoma.