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The effects of rapamycin on lens epithelial cell proliferation, migration, and matrix formation: an in vitro study.

Liu H, Feng G, Wu L, Fu S, Liu P, Yang W, Zhang X - Mol. Vis. (2010)

Bottom Line: The effect of rapamycin on the synthesis of Fn was examined via immunofluorescence.Extracellular matrix Fn formation of rLECs was also reduced by rapamycin.In our study, rapamycin strongly inhibited rLEC proliferation, bFGF-induced migration, and extracellular matrix Fn formation.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, The 1st Affiliated Hospital, Harbin Medical University, Harbin, PR China.

ABSTRACT

Purpose: The objective of the present study was to investigate the efficacy of rapamycin on rabbit lens epithelial cell proliferation, migration, and secretion of extracellular matrix fibronectin (Fn).

Methods: Rabbit lens epithelium cells (rLECs) were isolated from 1 month old rabbit. rLECs were either cultured for 24, 48, or 72 h with different doses of rapamycin (0.1, 1, and 10 ng/ml). The proliferation kinetics, proliferating cell nuclear antigen (PCNA) expression, and basic fibroblast growth factor (bFGF)-induced migration of rLEC was determined by methyl thiazol tetrazolium (MTT) assay, western blotting and transwell chamber assay, respectively. The effect of rapamycin on the synthesis of Fn was examined via immunofluorescence.

Results: Rapamycin significantly inhibited rLEC proliferation and PCNA protein expression when administered doses and time periods except for 0.1 ng/ml for 24 h. bFGF-induced migration rLECs was inhibited by pretreatment with rapamycin for 48 h. Extracellular matrix Fn formation of rLECs was also reduced by rapamycin.

Conclusions: In our study, rapamycin strongly inhibited rLEC proliferation, bFGF-induced migration, and extracellular matrix Fn formation. Thus, rapamycin may have a potential inhibition of posterior capsule opacification (PCO) and needs further study.

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Related in: MedlinePlus

Expression of Fn in rLECs. Expression of Fn and β-actin are analyzed in panel A. The Fn expression is standardized by an internal control (anti-β-actin). Lanes M, 1, 2, 3, 4, and 5 are protein marker, normal control, 0.1 ng/ml for 24 h, 0.1 ng/ml for 48 h, 10 ng/ml for 24 h, 10 ng/ml rapamycin for 48 h groups, respectively. The summary results of Fn is shown in panel B. All differences were statistically significant (*p≤0.05).
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f7: Expression of Fn in rLECs. Expression of Fn and β-actin are analyzed in panel A. The Fn expression is standardized by an internal control (anti-β-actin). Lanes M, 1, 2, 3, 4, and 5 are protein marker, normal control, 0.1 ng/ml for 24 h, 0.1 ng/ml for 48 h, 10 ng/ml for 24 h, 10 ng/ml rapamycin for 48 h groups, respectively. The summary results of Fn is shown in panel B. All differences were statistically significant (*p≤0.05).

Mentions: The effect of rapamycin on the expression of intracellular Fn was evaluated in vitro (Figure 6). The results showed intense staining of fibronectin without rapamycin incubation in comparison with rapamycin incubations. As shown in Figure 7, Fn expression in rLEC was examined by western blotting. rLECs without rapamycin treatment appeared more Fn expression when compared to the rapamycin incubations. Rapamycin, decreased approximately threefold at 10 ng/ml in comparison with the control cells. Taken together, the results showed that rapamycin significantly decreased Fn expression in rLECs.


The effects of rapamycin on lens epithelial cell proliferation, migration, and matrix formation: an in vitro study.

Liu H, Feng G, Wu L, Fu S, Liu P, Yang W, Zhang X - Mol. Vis. (2010)

Expression of Fn in rLECs. Expression of Fn and β-actin are analyzed in panel A. The Fn expression is standardized by an internal control (anti-β-actin). Lanes M, 1, 2, 3, 4, and 5 are protein marker, normal control, 0.1 ng/ml for 24 h, 0.1 ng/ml for 48 h, 10 ng/ml for 24 h, 10 ng/ml rapamycin for 48 h groups, respectively. The summary results of Fn is shown in panel B. All differences were statistically significant (*p≤0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2927374&req=5

f7: Expression of Fn in rLECs. Expression of Fn and β-actin are analyzed in panel A. The Fn expression is standardized by an internal control (anti-β-actin). Lanes M, 1, 2, 3, 4, and 5 are protein marker, normal control, 0.1 ng/ml for 24 h, 0.1 ng/ml for 48 h, 10 ng/ml for 24 h, 10 ng/ml rapamycin for 48 h groups, respectively. The summary results of Fn is shown in panel B. All differences were statistically significant (*p≤0.05).
Mentions: The effect of rapamycin on the expression of intracellular Fn was evaluated in vitro (Figure 6). The results showed intense staining of fibronectin without rapamycin incubation in comparison with rapamycin incubations. As shown in Figure 7, Fn expression in rLEC was examined by western blotting. rLECs without rapamycin treatment appeared more Fn expression when compared to the rapamycin incubations. Rapamycin, decreased approximately threefold at 10 ng/ml in comparison with the control cells. Taken together, the results showed that rapamycin significantly decreased Fn expression in rLECs.

Bottom Line: The effect of rapamycin on the synthesis of Fn was examined via immunofluorescence.Extracellular matrix Fn formation of rLECs was also reduced by rapamycin.In our study, rapamycin strongly inhibited rLEC proliferation, bFGF-induced migration, and extracellular matrix Fn formation.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, The 1st Affiliated Hospital, Harbin Medical University, Harbin, PR China.

ABSTRACT

Purpose: The objective of the present study was to investigate the efficacy of rapamycin on rabbit lens epithelial cell proliferation, migration, and secretion of extracellular matrix fibronectin (Fn).

Methods: Rabbit lens epithelium cells (rLECs) were isolated from 1 month old rabbit. rLECs were either cultured for 24, 48, or 72 h with different doses of rapamycin (0.1, 1, and 10 ng/ml). The proliferation kinetics, proliferating cell nuclear antigen (PCNA) expression, and basic fibroblast growth factor (bFGF)-induced migration of rLEC was determined by methyl thiazol tetrazolium (MTT) assay, western blotting and transwell chamber assay, respectively. The effect of rapamycin on the synthesis of Fn was examined via immunofluorescence.

Results: Rapamycin significantly inhibited rLEC proliferation and PCNA protein expression when administered doses and time periods except for 0.1 ng/ml for 24 h. bFGF-induced migration rLECs was inhibited by pretreatment with rapamycin for 48 h. Extracellular matrix Fn formation of rLECs was also reduced by rapamycin.

Conclusions: In our study, rapamycin strongly inhibited rLEC proliferation, bFGF-induced migration, and extracellular matrix Fn formation. Thus, rapamycin may have a potential inhibition of posterior capsule opacification (PCO) and needs further study.

Show MeSH
Related in: MedlinePlus