Downregulation of SMP30 in senescent human lens epithelial cells
Downregulation of SMP30 in senescent human lens epithelial cells
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衰老人晶状体上皮细胞中 SMP30 的下调
DOI：
10.3892/mmr.2017.7106
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发表时间：
2017-10-01
影响因子：
3.4
通讯作者：
Liang, Hao
中科院分区：
文献类型：
--
作者：
Li, Shuning;Chen, Xi;Liang, Hao
据报道，衰老标志蛋白30 （SMP30）具有抗凋亡和抗氧化作用，并参与Ca2+调节，可能参与白内障的发生和发展。本研究旨在研究SMP30在衰老人晶状体上皮细胞（HLECs）中的表达，并探讨SMP30与衰老的关系。采用H2O2模拟HLEC细胞系SRA01/04细胞衰老，显微镜下观察细胞形态学变化，MTT法、衰老相关-β-半乳糖苷酶（SA-β-Gal）染色及细胞周期分析检测细胞活性。采用逆转录-定量聚合酶链反应（RT-qPCR）和western blotting检测SMP30 mRNA和蛋白的表达。长期低剂量H2O2暴露后，细胞表现出衰老相关的形态学改变，生长活性降低，SA-β-Gal阳性染色增加，细胞周期阻滞在S期和G2/M期。暴露于75µM和100µM H2O2后，SMP30 mRNA表达水平显著下调，与对照组相比，同一组细胞中SMP30 mRNA表达水平降低了6倍。然而，在25µM和50µM H2O2暴露组中，SMP30的表达没有明显变化。这些结果表明，在h2o2介导的慢性氧化应激诱导衰老的早期，SMP30的表达可能没有明显变化，但当氧化应激增加、衰老加剧时，SMP30可能在衰老的HLECs中显著下调。本研究提示SMP30可能是参与高晶状体老化过程和白内障发生的重要因子。
Senescence marker protein 30 (SMP30) has been reported to serve antiapoptotic and antioxidant roles, as well as roles in Ca2+ regulation, and may be involved in the occurrence and development of cataract. The present study aimed to investigate the expression of SMP30 in senescent human lens epithelial cells (HLECs) and explored the relationship between SMP30 and aging. SRA01/04 cells, a HLEC line, were treated with H2O2 to mimic aging, and cell morphological changes were observed by microscopy and cell activity was examined by MTT assay, senescence-associated-β-galactosidase (SA-β-Gal) staining and cell cycle analysis. The expression of SMP30 mRNA and protein was measured by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blotting. Following prolonged low-dose H2O2 exposure, cells exhibited senescence-related morphological changes, reduced growth activity, increased SA-β-Gal positive staining and cell cycle arrest in the S and G2/M phases. SMP30 mRNA expression levels were significantly downregulated following exposure to 75 and 100 µM H2O2, and the protein expression levels in the same groups were decreased by >6-fold compared with the control untreated cells. However, no significant change was observed in SMP30 expression in the 25 and 50 µM H2O2 exposure groups. These results suggest that, in the early stage of senescence induced by H2O2-mediated chronic oxidative stress, there may be no significant change in SMP30 expression, but when the oxidative stress increases and senescence is aggravated, SMP30 may be significantly downregulated in the senescent HLECs. The present study indicates that SMP30 may be an important factor involved in the aging process of HLECs and the development of cataract.