[Preliminary study of inhibition of human Tenon's capsule fibroblasts in vitro by RNA interference targeting IKK-beta].
OBJECTIVE
To determine whether small interference RNA (siRNA) targeting the inhibitor of kappaB kinase-beta (IKK-beta) could be used to suppress the IKK-beta expression, and inhibit the proliferation of human Tenon's capsule fibroblasts in vitro.


METHODS
IKK-beta specific siRNA designed from the human gene sequence was transfected into the cultured human Tenon's capsule fibroblasts, and a non-targeted siRNA was transfected as a negative control. The mRNA of IKK-beta was assessed by reverse transcription-polymerase chain reaction (RT-PCR), and the protein expression was determined by Western blotting. Cell viability of the cultured human Tenon's capsule fibroblasts with series of RNA interference at 5, 10, 25, 50, 100, and 200 nmol/L was evaluated by MTT assay 48 hours after transfection.


RESULTS
The expression of IKK-beta was significantly (P < 0.05) suppressed at both mRNA and protein levels after transfection. The proliferation of the cultured human Tenon's capsule fibroblasts was inhibited at all the transfected concentrations at different rates (10.72%, 23.35%, 30.84%, 51.25%, 50.06% and 49.63% respectively). The highest level of inhibition was observed at 50 nmol/L of siRNA concentration.


CONCLUSIONS
IKK-beta specific siRNA is effective in suppressing the IKK-beta expression and inhibiting the proliferation of the cultured human Tenon's capsule fibroblasts. RNA interference may offer a new alternative to post-operational management of scar tissue formation.
Huang
Sheng-song
SS
Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangzhou 510060, China.
Ge
Jian
J
Wang
Li-na
LN
Yin
Xin-bao
XB
Wei
Yan-tao
YT
Ma
Ping
P
D000328:Adult; D049109:Cell Proliferation; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D005902:Glaucoma, Open-Angle / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D051550:I-kappa B Kinase / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering; D014162:Transfection
D012333:RNA, Messenger; D034741:RNA, Small Interfering; D051550:I-kappa B Kinase