An antisense plasmid targeting survivin expression induces apoptosis and sensitizes hepatocarcinoma cells to chemotherapy.
- Author:
Wanguang ZHANG
1
;
Xiaoping CHEN
;
Fazu QIU
Author Information
1. Hepatic Surgery Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030.
- Publication Type:Journal Article
- MeSH:
Antigens, Neoplasm;
biosynthesis;
genetics;
Antimetabolites, Antineoplastic;
pharmacology;
Apoptosis;
Carcinoma, Hepatocellular;
metabolism;
pathology;
Cell Line, Tumor;
DNA, Antisense;
Fluorouracil;
pharmacology;
Humans;
Inhibitor of Apoptosis Proteins;
Liver Neoplasms;
metabolism;
pathology;
Microtubule-Associated Proteins;
biosynthesis;
genetics;
Neoplasm Proteins;
Plasmids;
RNA, Messenger;
biosynthesis;
genetics;
Transfection
- From:
Journal of Huazhong University of Science and Technology (Medical Sciences)
2003;23(4):387-391
- CountryChina
- Language:English
-
Abstract:
To explore the change of sensitivity to chemotherapy of antisense RNA targeting survivin on hepatocarcinoma carcinoma cells in vitro. Survivin mRNA structure region was amplified by RT-PCR and inserted inversely into eukaryotic expression vector pcDNA3. The antisense expression plasmid pcDNA3/survivin was transfected into HepG2 with lipofectAMINE 2000 (LF2000), with low concentration of 5-fluorouracil (5-Fu) added. Survivin protein was detected by Western-blot, the growth activity was measured by MTT, and apoptosis was detected by Flow Cytometry 12 h, 24 h, 48 h after transfection. The activity of caspase-3 was found by quantitative assay 48 h after transfection. The construction of antisense RNA vector pcDNA3/survivin was verified by restricted endonuclease digestion and nucleotide sequencing. Compared with normal group, 5-Fu and antisense survivin group, the cells growth inhibition, apoptosis index, and caspase-3 activity were increased in antisense survivin transfected + 5-Fu group. The threshold of apoptosis was decreased after survivin was silenced, and the sensitivity to chemotherapy was increased. These findings suggest the existence of a potential new target for gene therapy.
