HCCR-2基因沉默后HepG2肝癌细胞形态变化

The Morphology of HepG2 Liver Cancer Cells after Silencing HCCR-2 Gene

目的研究HCCR-2基因表达沉默后肝癌细胞HepG2形态学的变化.方法将经测序证实的干扰质粒用脂质体法转染HepG2细胞,经G418持续压力选择和有限稀释法获得稳定转染的细胞系.分别用光镜、HE染色、荧光显微镜、透射电镜观察HepG2细胞转染前后形态学的改变.结果与亲本细胞相比,HCCR-2沉默后,HepG2细胞呈梭形,核浆比例有所下降,透射电镜下可观察到细胞核收缩变圆,胞质浓缩,细胞核中染色质边集、皱缩,密度增高.结论 RNAi介导的HCCR-2基因沉默可以促进HepG2细胞形态的改变及凋亡.

Objective To investigate the morphological changes of HepG2 after silencing HCCR-2 by RNAi.Methods After confirmed by sequencing,the recombinant HCCR-2 RNAi vector was transfected into HepG2 cells by LipofectamineTM2000.Cells containing stable transformants were selected by the ability of resistance to G418 and isolated with a limited dilution.Morphological changes of the transfected cells were examined under light microscope,HE staining,inverted,fluorescence microscope and transmission electron microscope respectively.Results The transfected cells were transformed into fusiform and katyoplasmic ratio decreased to some extent.Cellular nucleus contracted and became round,cell kytoplasm condensed in the transfected cells.Chromatin margination and crimple were also observed under transmission electron in the transfected cells.Conclusion RNAi eukaryotic expression vector targeting HCCR-2 can promote the morphological changes and apoptosis of HepG2 cells