预知子种子提取物诱导肝癌细胞内质网应激的形态学特征

Akebia Fruit seed extract induced morphological characteristics of endoplasmic reticulum stress in hepatocellular carcinoma cells

目的观察预知子种子提取物对不同肝癌细胞形态学的影响。方法综合采用细胞生物学和扫描电子显微镜、透射电子显微镜等技术,观察预知子种子乙醇提取物干预后HepG2、Huh 7、SMMC 7721等肝癌细胞的形态学特征。结果 HepG2呈重叠生长,表面布满绒毛;预知子种子处理后部分细胞绒毛萎缩、消失,细胞形态呈不规则圆,或有塌陷,糙面内质网出现不同程度的扩张。Huh 7细胞会自发形成凋亡;预知子种子处理后细胞表面绒毛变得不规则、萎缩,多见凋亡小体,细胞中可见大量内质网应激所形成的空泡。SMMC 7721细胞表面布满较长绒毛;预知子种子处理后多见形态异常,胞体膨胀,呈不规则圆,细胞表面部分区域隆起或塌陷,一些细胞表面因严重内质网应激而发生塌陷,胞内多见大量严重扩张的内质网。结论 HepG2、Huh 7、SMMC 7721等肝癌细胞各具形态学特征,经预知子种子提取物给药后均不同程度地诱导出严重的内质网应激,这可能是预知子种子治疗肝癌的作用途径之一。

Objective To observe the effects of Akebia Fruit seed （AFS） extract on morphology of different hepatocellular carcinoma cells. Methods By the comprehensive application of cell biology, scanning electron microscopy（SEM） and transmission electron microscopy（ TEM）, the effects of alcohol extract of AFS on the morphological characteristics of HepG2, Huh 7, and SMMC7721 were observed. Results Untreated HepG2 cells show overlapping growth and the surfaces are fully covered with villus. After the treatment with AFS extract, the partial cell villus atrophied and disappeared, the cells became irregular and even collapsed, and rough endoplasmic reticulum（ER） expanded to different degrees. Spontaneous apoptosis could form in Huh 7 cells. After the treatment with AFS extract, the irregular villus, cell shrinking, a lot of apoptotic bodies, and numerous vacuoles caused by endoplasmic reticulum stress were observed in cells. Untreated SMMC- 7721 cells are fully covered with longer villus. After the treatment with AFS extract, abnormal morphology could be detected. The cell body expansion with irregular round shape were observed, some regions on the surface of cells collapsed or upheaved, the surface of some cells collapsed due to severe endoplasmic reticulum stress, and a large number of heavily expanded endoplasmic reticulum were commonly found. Conclusion These three kinds of hepatocellular carcinoma cell lines exhibit various morphological characteristics, but serious ER stress with different degrees can be induced after the treatment with Akebia Fruit seed extract, which may be one way of Akebia Fruit seed acting on liver cancer.