Vector-mediated shRNA inhibits HIF-1alpha expression in prostate cancer cells / 中华男科学杂志
National Journal of Andrology
; (12): 993-997, 2008.
Article
ي Zh
| WPRIM
| ID: wpr-309772
المكتبة المسؤولة:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To construct a short hairpin RNA (shRNA) vector of the hypoxia inducible factor-1alpha (HIF-1alpha), determine its inhibitory effect on the expression of the HIF-1alpha gene in PC-3M cells, and investigate its application prospects in the treatment of prostate cancer.</p><p><b>METHODS</b>We designed and synthesized the shRNA template sequence specific against HIF-lalpha, inserted it into the vector psilencer 2.1-U6 to generate the plasmid psilencer-HIF, transfected the recombinant plasmid into prostate cancer cell line PC-3M cells and detected the transfection efficiency by cotransfection with the pEGFP vector as well as the expression of HIF-1alpha by RT-PCR and Western blot.</p><p><b>RESULTS</b>The DNA sequencing analysis showed a complete consistency of the recombinant plasmid psilencer-HIF with the design. Twenty-four hours after the transfection, the rate of transfected plasmid was about (89.26 +/- 4.72)% and the vector-mediated shRNA induced RNA interference (RNAi), while 48 hours transfection reduced the HIF-1alpha mRNA and protein levels by 82.09% and 81.61% respectively (P < 0.01) in PC-3M cells.</p><p><b>CONCLUSION</b>The shRNA vector was successfully constructed, which can effectively suppress the expression of HIF-1alpha in prostate cancer cells.</p>
النص الكامل:
1
الفهرس:
WPRIM
الموضوع الرئيسي:
Prostatic Neoplasms
/
RNA, Messenger
/
Base Sequence
/
Transfection
/
Gene Expression
/
RNA, Small Interfering
/
Cell Line, Tumor
/
Hypoxia-Inducible Factor 1, alpha Subunit
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Genetic Vectors
/
Genetics
المحددات:
Humans
/
Male
اللغة:
Zh
مجلة:
National Journal of Andrology
السنة:
2008
نوع:
Article