[Vector-mediated shRNA inhibits HIF-1alpha expression in prostate cancer cells].
OBJECTIVE
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.


METHODS
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.


RESULTS
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.


CONCLUSIONS
The shRNA vector was successfully constructed, which can effectively suppress the expression of HIF-1alpha in prostate cancer cells.
Ping
Hao
H
Department of Urology, Beijing Chaoyang Hospital, Capital University of Medical Sciences, Beijing 100020, China. pinghaozx@yahoo.com.cn
Xing
Nian-Zeng
NZ
Chen
Xiao-Chun
XC
Zhang
Jun-Hui
JH
Yan-Yong
Yang
Yong
Y
D001483:Base Sequence; D045744:Cell Line, Tumor; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics*; D008297:Male; D011471:Prostatic Neoplasms / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering; D014162:Transfection
D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D012333:RNA, Messenger; D034741:RNA, Small Interfering