肝癌细胞自噬性凋亡过程中caspase-3基因的克隆与鉴定

目的从长春新碱诱导发生自噬性凋亡的肝癌细胞系HepG2中克隆caspase-3基因。方法提取细胞总RNA,采用RT-PCR技术扩增目的片段,构建重组克隆载体,测序鉴定及分析。结果经RT-PCR获得了预期的扩增产物即caspase-3基因625bp的核苷酸片段,其测序结果与文献报道一致。结论初步证明该caspase-3基因片段参与了长春新碱诱导的HepG2细胞自噬性凋亡过程。

Cloning and Identification of Caspase-3 Gene from HepG2 Cells that Underwent Autophagic Apoptosis

Objective To acquire the fragment of caspase-3 gene from HepG2 cells, a human hepatocarcinoma cell line,that underwent autophagic apoptosis induced by vincristine(VCR).Methods Total RNA was extracted from the HepG2 cell line,then the fragment of caspase-3 gene from the cells was amplified by RT-PCR and it was cloned,sequenced by gene recombination techniques for confirmation.Results The prospective sequence of selected cloned caspase-3 gene was 625 bp.It is consistent with that of reported human caspase-3 gene sequence.Conclusions It was proved that the fragment of caspase-3 gene is involved in autophagic apoptosis induced by VCR on HepG2 cells.