Hong-wei Liu1, Xiang-lan Wang, Shu-ling Peng, Yi Zhou, Dong-ling Liu. 1. Department of Pharmacology, Beijing Institute of Ophthalmology, Beijing Tongren Ophthalmology Center, Capital University of Medical Sciences, Beijing 100005, China. hw65@sohu.com
Abstract
OBJECTIVE: To evaluate the effects of basic fibroblast growth factor (bFGF) antisense oligonucleotides and their liposomes on the proliferation of cultured rat lens epithelial cells (LECs). METHODS: The rat LECs were cultured and bFGF antisense oligonucleotides (AONs), sense oligonucleotides (SONs), AON liposomes, SON liposomes were added to second passage of cells supplied with DMEM while empty liposomes served as controls. MTT assay was used to examine the proliferation of LECs, and RT-PCR was performed to quantify bFGF mRNA expression in LECs 24 h after treatment. RESULTS: The Shapes of rat LECs were polygonal after 24 h in culture. Cell confluence was reached in 2 - 3 weeks. After subculture, confluence was occurred in 1 - 2 weeks. The results of MTT assay were showed that the absorption (A) value of bFGF AONs was 0.138 +/- 0.074, and that of bFGF SONs and DMEM control were 0.325 +/- 0.097 and 0.370 +/- 0.079, respectively. The A value of AONs was significantly less than SONs and DMEM alone (P = 0.024, P = 0.005). The absorption (A) value of bFGF AON liposomes was 0.128 +/- 0.032, and that of bFGF SON liposomes and liposomes alone were 0.238 +/- 0.120 and 0.348 +/- 0.017. The absorption (A) value of AON liposomes was not significantly different from that of liposomes negative control (P = 0.000). By RT-PCR, the amounts of PCR product for bFGF AONs, SONs and DMEM control were 0.33 micro g, 0.99 micro g, 0.85 micro g. The amounts of PCR product of bFGF AON liposomes, SON liposomes and liposomes only were 0.23 micro g, 0.48 micro g, 0.56 micro g. CONCLUSIONS: The proliferation of cultured rat LECs is inhibited by the treatment of the antisense oligonucleotides, the inhibition is correlated with decreased expression of bFGF mRNA.
OBJECTIVE: To evaluate the effects of basic fibroblast growth factor (bFGF) antisense oligonucleotides and their liposomes on the proliferation of cultured rat lens epithelial cells (LECs). METHODS: The rat LECs were cultured and bFGF antisense oligonucleotides (AONs), sense oligonucleotides (SONs), AON liposomes, SON liposomes were added to second passage of cells supplied with DMEM while empty liposomes served as controls. MTT assay was used to examine the proliferation of LECs, and RT-PCR was performed to quantify bFGF mRNA expression in LECs 24 h after treatment. RESULTS: The Shapes of rat LECs were polygonal after 24 h in culture. Cell confluence was reached in 2 - 3 weeks. After subculture, confluence was occurred in 1 - 2 weeks. The results of MTT assay were showed that the absorption (A) value of bFGFAONs was 0.138 +/- 0.074, and that of bFGF SONs and DMEM control were 0.325 +/- 0.097 and 0.370 +/- 0.079, respectively. The A value of AONs was significantly less than SONs and DMEM alone (P = 0.024, P = 0.005). The absorption (A) value of bFGF AON liposomes was 0.128 +/- 0.032, and that of bFGF SON liposomes and liposomes alone were 0.238 +/- 0.120 and 0.348 +/- 0.017. The absorption (A) value of AON liposomes was not significantly different from that of liposomes negative control (P = 0.000). By RT-PCR, the amounts of PCR product for bFGFAONs, SONs and DMEM control were 0.33 micro g, 0.99 micro g, 0.85 micro g. The amounts of PCR product of bFGF AON liposomes, SON liposomes and liposomes only were 0.23 micro g, 0.48 micro g, 0.56 micro g. CONCLUSIONS: The proliferation of cultured rat LECs is inhibited by the treatment of the antisense oligonucleotides, the inhibition is correlated with decreased expression of bFGF mRNA.