Construction of miRNA expression vector dual-targeting on HIF-1α/survivin genes and its effects on proliferation of pancreatic cancer Panc-1 cell line / 浙江大学学报·医学版
Journal of Zhejiang University. Medical sciences
;
(6): 81-88, 2012.
Article
in Chinese
| WPRIM
| ID: wpr-247179
ABSTRACT
<p><b>OBJECTIVE</b>To design and construct miRNA expression vector dual-targeting on HIF-1α and survivin genes and to investigate its effects on proliferation of human pancreatic cancer cells.</p><p><b>METHODS</b>The specific pre-miRNA single strand DNA oligos for HIF-1 α and survivin genes were designed and synthesized, then via annealing and ligating with pcDNA6.2-GW/EmGFP-miR plasmids in order, two kinds (eight in total) of miRNA expression vectors for HIF-1α and survivin genes were constructed. The vectors, which were most effective to knockdown target genes, were screened with real-time RT-PCR and combined by chaining technology to construct dual-targeting plasmid. The recombined dual-targeting plasmid, mono-targeting plasmids and negative plasmid were transfected into Panc-1 cells, the suppression effect on two genes was identified by real-time RT-PCR, Western blot and MTT assays.</p><p><b>RESULTS</b>The miRNA expression plasmids anti-H, anti-S and anti-H+S were successfully constructed by identification of sequencing analysis, and they were able to effectively inhibit the target genes expressing. MTT assays showed that the inhibition effect of dual-targeting vector anti-H+S was higher than that of mono-targeting vectors anti-H and anti-S 72 h after transfection (P<0.05).</p><p><b>CONCLUSION</b>The effective miRNA expression vector dual-targeting on HIF-1α and survivin genes has been successfully constructed. The inhibition effect on proliferation of pancreatic cancer Panc-1 cells by dual-targeting plasmid was higher than that by mono-target plasmids.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pancreatic Neoplasms
/
Pathology
/
Plasmids
/
Transfection
/
MicroRNAs
/
Cell Line, Tumor
/
Cell Proliferation
/
Inhibitor of Apoptosis Proteins
/
Hypoxia-Inducible Factor 1, alpha Subunit
/
Genetic Vectors
Type of study:
Prognostic study
Limits:
Humans
Language:
Chinese
Journal:
Journal of Zhejiang University. Medical sciences
Year:
2012
Type:
Article
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