[Construction of RNA recombinant vector expressing STAT3 short hairpin and its effects on proliferation and apoptosis of SiHa cells].
OBJECTIVE
To investigate the effects of eukaryotic vector expressing short hairpin RNA (shRNA) of signal transducers and activators of transcription 3 (STAT3) on the proliferation and apoptosis of SiHa cells.


METHODS
shRNA templates were designed based on STAT3 gene sequence and cloned into pSilencer2.1-U6-neo vector. The resultant plasmid was transfected into SiHa cells with Lipofectamine 2000. The STAT3 protein and mRNA were detected by Western blot and RT-PCR, respectively. The cellular growth activity was assayed by MTT and the apoptosis was tested by flow cytometry.


RESULTS
The plasmid pSilencer2.1-U6-neo-STAT3 was successfully constructed and transfected into SiHa cells. The expression of STAT3 in SiHa cells decreased, the cellular growth activity decreased, and the cell apoptosis increased.


CONCLUSIONS
The siRNA expressing plasmid pSilencer2.1-U6-neo-STAT3 can inhibit cellular proliferation and induce the apoptosis of cervical cancer cells by suppressing the expression of STAT3.
Wu
Wei-Guang
WG
Department of Obstetrics & Gynecology, Affiliated Hospital, Medical College of Chinese People's Armed Police Forces, Tianjin 300162, China.
Yu
Yue-Cheng
YC
Chen
Ya-Qiong
YQ
Cao
Xiu-Qin
XQ
D017209:Apoptosis / Q000235:genetics / Q000502:physiology*; D015153:Blotting, Western; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005260:Female; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000235:genetics / Q000502:physiology*; D006801:Humans; D012313:RNA / Q000235:genetics / Q000502:physiology*; D034741:RNA, Small Interfering / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050796:STAT3 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D002583:Uterine Cervical Neoplasms / Q000378:metabolism / Q000628:therapy
D034741:RNA, Small Interfering; C039919:RNA, recombinant; D050796:STAT3 Transcription Factor; D012313:RNA