Transfection of CTGF siRNA inhibits transdifferentiation in human lens epithelium cell line B3 in vitro
10.3980/j.issn.1672-5123.2017.8.01
- VernacularTitle:CTGF siRNA转染对体外培养人晶状体上皮细胞株B3转分化的抑制作用
- Author:
Hua, ZHUANG
;
Ning-Xuan, ZHENG
;
Jing, WU
;
Wei, XU
;
Jian-Zhang, HU
;
Mao-Song, XIE
;
Jian, GUO
;
Guo-Xing, XU
- Keywords:
connective tissue growth factor;
small interfering RNA;
lipofectamine;
transdifferentiation
- From:
International Eye Science
2017;17(8):1387-1393
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To investigate the expression of connective tissue growth factor (CTGF) and α-SMA in human lens epithelium cell (HLEC) line B3 after transfection by liposome-coated siRNA targeting CTGF.METHODS: HLECs were transfected with small interfering RNA (siRNA) targeting CTGF,labeled with 5`-fluorescein isothiocyanate (5`-FITC) and coated with lipofectamine.The transfection ratio was evaluated via fluorescence intensity.Cell counting kit-8 (CCK-8) assay was performed to assess cytoviability of both non-transfected and transfected HLECs.Quantitative RT-PCR,cell immunochemistry and Western blot analysis were conducted to detect the expression changes of CTGF and α-SMA after transfection.RESULTS: A highly effective transfection ratio was observed in siRNA co-transfected with lipofectamine.The transfection ratio reached 95% at 24h.The proliferation of HLECs was inhibited by siRNA after 72h transfection.The expression of CTGF and α-SMA significantly decreased in HLECs after transfected by CTGF siRNA for 24h.This effect was not found in negative control siRNA.CONCLUSIONS: SiRNA targeting CTGF decreased CTGF and α-SMA expression in HLECs,which is a potential therapeutic strategy for posterior capsular opacification.
