Optical redox ratio differentiates breast cancer cell lines based on estrogen receptor status.
Optical redox ratio differentiates breast cancer cell lines based on estrogen receptor status.
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DOI：
10.1158/0008-5472.can-09-2572
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发表时间：
2010-06-01
期刊：
影响因子：
11.2
通讯作者：
Ramanujam N
中科院分区：
文献类型：
--
作者：
Ostrander JH;McMahon CM;Lem S;Millon SR;Brown JQ;Seewaldt VL;Ramanujam N
自体荧光光谱是一种利用内源荧光团的强大成像技术。内源性荧光团 NADH 和黄素腺嘌呤二核苷酸 (FAD) 分别是细胞代谢中两个主要的电子供体和受体。光氧化还原 (redox) 比率是细胞代谢的量度，可以通过 NADH/FAD 的比率来确定。我们假设，与乳腺肿瘤细胞系相比，正常乳腺上皮细胞的光学氧化还原比存在显着差异，并且雌激素受体（ER）阳性细胞比ER阴性细胞具有更高的氧化还原比。为了检验我们的假设，通过共焦显微镜收集 NADH 和 FAD 的荧光发射来确定光学氧化还原比。我们观察到与正常细胞系相比，癌症的光学氧化还原比有统计学上显着的增加（P < 0.05）。此外，我们观察到 ER(+) 乳腺癌细胞系的光学氧化还原比在统计上显着增加。通过实时 PCR 测定的 ESR1 表达水平与光学氧化还原比直接相关（Pearson 相关系数 = 0.8122，P = 0.0024）。此外，用他莫昔芬和 ICI 182,870 治疗仅在 ER(+) 乳腺癌细胞系中统计上降低了光学氧化还原比。这项研究的结果提出了一种重要的可能性，即荧光光谱可用于根据受体状态识别乳腺癌亚型、监测治疗反应或潜在预测治疗反应。这种光学对比源可能是临床前模型中药物筛选的潜在有用工具。
Autofluorescence spectroscopy is a powerful imaging technique that exploits endogenous fluorophores. The endogenous fluorophores NADH and flavin adenine dinucleotide (FAD) are two of the principal electron donors and acceptors in cellular metabolism, respectively. The optical oxidation-reduction (redox) ratio is a measure of cellular metabolism and can be determined by the ratio of NADH/FAD. We hypothesized that there would be a significant difference in the optical redox ratio of normal mammary epithelial cells compared with breast tumor cell lines and that estrogen receptor (ER)–positive cells would have a higher redox ratio than ER-negative cells. To test our hypothesis, the optical redox ratio was determined by collecting the fluorescence emission for NADH and FAD via confocal microscopy. We observed a statistically significant increase in the optical redox ratio of cancer compared with normal cell lines (P < 0.05). Additionally, we observed a statistically significant increase in the optical redox ratio of ER(+) breast cancer cell lines. The level of ESR1 expression, determined by real-time PCR, directly correlated with the optical redox ratio (Pearson’s correlation coefficient = 0.8122, P = 0.0024). Furthermore, treatment with tamoxifen and ICI 182,870 statistically decreased the optical redox ratio of only ER(+) breast cancer cell lines. The results of this study raise the important possibility that fluorescence spectroscopy can be used to identify subtypes of breast cancer based on receptor status, monitor response to therapy, or potentially predict response to therapy. This source of optical contrast could be a potentially useful tool for drug screening in preclinical models.