Laminins in an in vitro anterior lens capsule model established using HLE B-3 cells

Yan,Yu; Qian,Hua; Jiang,Hongda; Yu,Haiyang; Sun,Liyao; Wei,Xi; Sun,Yunduan; Ge,Hongyan; Zhou,Haizhou; Li,Xiaoguang; Hashimoto,Takashi; Tang,Xianling; Liu,Ping

doi:10.3892/mmr.2018.8581

Laminins in an in vitro anterior lens capsule model established using HLE B-3 cells

Authors:
- Yu Yan
- Hua Qian
- Hongda Jiang
- Haiyang Yu
- Liyao Sun
- Xi Wei
- Yunduan Sun
- Hongyan Ge
- Haizhou Zhou
- Xiaoguang Li
- Takashi Hashimoto
- Xianling Tang
- Ping Liu
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Affiliations: Eye Hospital, First Affiliated Hospital, Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China, Academician Workstation, Harbin Medical University and Heilongjiang Academy of Medical Sciences, Harbin, Heilongjiang 150081, P.R. China, Department of Laboratory Diagnosis, First Affiliated Hospital, Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China, Department of Dermatology, Osaka City University Graduate School of Medicine, Osaka 545‑8585, Japan
Published online on: February 8, 2018 https://doi.org/10.3892/mmr.2018.8581
Pages: 5726-5733
Copyright: © Yan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Cataracts are the most common eye disease to cause blindness in patients. The abnormal deposition of laminins (LMs) in the lens capsule and the disruption of capsular epithelium contribute to cataract development, although the mechanism by which this occurs is currently unclear. The present study aimed to reproduce HLE B‑3 basement membranes (BMs) using HLE B‑3 cells and to analyze the similarities of LM expression between HLE B‑3 BMs and human anterior lens capsule (ALC). Immunohistochemistry (IHC), ELISA, western blot analysis and immunoprecipitation (IP)‑western blot analysis were used to detect total LMs, LM trimers and 11 LM subunits in HLE B‑3 cells, HLE B‑3 BMs and human ALCs. In IHC staining, HLE B‑3 cells and human ALCs were positive for LMs. In LM ELISA, all samples analyzed were positive for LMs. Western blot analysis detected all LM subunits except for LMγ3 in HLE B‑3 cell lysate, 4 subunits (LMα4, LMα2, LMα1 and LMγ1) in HLE B‑3 cell culture supernatant, 5 subunits (LMα4, LMα2, LMα1, LMβ3 and LMγ1) in HLE B‑3 BMs, and 3 subunits (LMα4, LMγ2 and LMγ1) in human ALCs. The results of IP‑western blot analysis revealed that the LM411 trimer was detected in HLE B‑3 cell culture supernatant. These results indicated that HLE B‑3 BMs were similar to human ALCs in terms of LM expression. Therefore, HLE B‑3 BMs could be used as an in vitro ALC model to determine the role of LMs in ALC in the pathogenesis of cataracts and to select potential anti‑cataract drugs.

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