Bains Savneet Kaur, Chapman Kim, Bright Scott, Senan Anish, Kadhim Munira, Slijepcevic Predrag
a Department of Life Sciences , Brunel University London , Uxbridge , UK.
b Department of Biological and Medical Sciences , Oxford Brookes University , Oxford , UK.
Int J Radiat Biol. 2019 Jan;95(1):54-63. doi: 10.1080/09553002.2018.1466066. Epub 2018 May 3.
To investigate the effects of ionizing radiation on telomere length and telomerase activity in human lens epithelial cells. There are studies suggesting evidence of telomere length in association with opacity of the lens; however, these studies have been conducted on Canine Lens cells. Our study was designed to understand further the effects of different doses of ionizing radiation on telomere length and telomerase activity in cultured human lens epithelium cells from three Donors.
For this study, embryonic human lens epithelial (HLE) cells from three donors, obtained commercially were cultured. Telomere length and telomerase activity were measured after each passage until cells stopped growing in culture. This was repeated on irradiated (0.001 Gy, 0.01 Gy, 0.02 Gy, 0.1 Gy, 1 Gy and 2 Gy) cells. DNA damage response using the H2AX and telomere dysfunction foci assays were also examined at 30 mins, 24 hours, 48 hours and 72 hours postirradiation.
We have demonstrated genetic changes in telomere length and oxidative stress, which may be relevant to cataractogenesis. Our study shows that in control cells telomere length increases as passage increases. We have also demonstrated that telomere length increases at higher doses of 1.0 Gy and 2.0 Gy. However, telomerase activity decreases dose dependently and as passages increase. These results are not conclusive and further studies ex vivo measuring lens opacity and telomere length in the model would be beneficial in a bigger cohort, hence confirming a link between telomere length, cataractogenesis and genetic factors.
研究电离辐射对人晶状体上皮细胞端粒长度和端粒酶活性的影响。有研究表明存在端粒长度与晶状体混浊相关的证据;然而,这些研究是在犬晶状体细胞上进行的。我们的研究旨在进一步了解不同剂量的电离辐射对来自三名供体的培养人晶状体上皮细胞中端粒长度和端粒酶活性的影响。
在本研究中,培养从商业渠道获得的来自三名供体的人胚胎晶状体上皮(HLE)细胞。在每次传代后测量端粒长度和端粒酶活性,直至细胞在培养中停止生长。对经辐照(0.001 Gy、0.01 Gy、0.02 Gy、0.1 Gy、1 Gy和2 Gy)的细胞重复此操作。还在辐照后30分钟、24小时、48小时和72小时使用H2AX和端粒功能障碍灶检测法检测DNA损伤反应。
我们已经证明了端粒长度的遗传变化和氧化应激,这可能与白内障形成有关。我们的研究表明,在对照细胞中,端粒长度随着传代次数的增加而增加。我们还证明,在1.0 Gy和2.0 Gy的较高剂量下,端粒长度增加。然而,端粒酶活性呈剂量依赖性降低且随着传代次数增加而降低。这些结果并不确凿,在更大的队列中对该模型进行离体测量晶状体混浊和端粒长度的进一步研究将是有益的,从而证实端粒长度、白内障形成和遗传因素之间的联系。