An antisense plasmid targeting survivin expression induces apoptosis and sensitizes hepatocarcinoma cells to chemotherapy / 华中科技大学学报(医学)(英德文版)
Journal of Huazhong University of Science and Technology (Medical Sciences)
; (6): 387-391, 2003.
Article
in En
| WPRIM
| ID: wpr-330898
Responsible library:
WPRO
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.
Full text:
1
Index:
WPRIM
Main subject:
Pathology
/
Pharmacology
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Plasmids
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RNA, Messenger
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Transfection
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DNA, Antisense
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Apoptosis
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Carcinoma, Hepatocellular
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Cell Line, Tumor
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Inhibitor of Apoptosis Proteins
Limits:
Humans
Language:
En
Journal:
Journal of Huazhong University of Science and Technology (Medical Sciences)
Year:
2003
Type:
Article