[Inhibition of integrin beta1 expression in human keratinocyte stem cells by vector-1 based interfering RNA strategy].
OBJECTIVE
To investigate the influence of integrin beta1 on the proliferation and differentiation of human keratinocyte stem cells (KSCs).


METHODS
DNA oligonucleotides targeting integrin beta1 at different locations were synthesized and inserted into BamHI-1 HindIII linearized p Silencer 3.1/H1 plasmids. The inserted sequences were verified by DNA sequencing. The KSCs were divided into control (without transfection), T1 (with transfection of vacant vector), T2 (with transfection of si integrin beta(1-1) vector), T3 (with transfection of si integrin beta(1-1) vector), and T4 (with transfection of si Negative vector) groups. The change in the expression of integrin beta1, was determined with Western blotting. The positive vector with the highest expression of integrin beta1 was selected and named as integrin beta1, and semi-quantitative RT-PCR was employed to detect the change in the expression of integrin beta1 mRNA.


RESULTS
The protein expression of integrin beta1, was not suppressed in control and T1 group, but it was suppressed in T2 and T3 groups, especially in T3 group (the suppression rate was 60%-70%, which was named si integrin beta1). The expression of integrin beta1 mRNA was obviously decreased by integrin beta1, transfection (the suppression rate was 70%).


CONCLUSIONS
The expression of integrin beta1, mRNA and protein could be down-regulated with recombinant si integrin P, vector transfection.
Tang
Hong-ying
HY
Institute of Burn Research, Southwest Hospital, State Key Laboratory of Trauma, Burns and Combined Injury, Third Military Medical University, Chongqing 400038, PR China.
Wu
Su-hua
SH
Su
Yong-yue
YY
Chen
Jian
J
Liu
Yue-ming
YM
Liang
Guang-ping
GP
Sun
Rong-ju
RJ
Luo
Xiang-dong
XD
D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D005822:Genetic Vectors; D006801:Humans; D019012:Integrin beta1 / Q000235:genetics / Q000378:metabolism*; D015603:Keratinocytes / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D013234:Stem Cells / Q000378:metabolism*; D014162:Transfection
D019012:Integrin beta1; D012333:RNA, Messenger; D034741:RNA, Small Interfering