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拉曼光谱学:一种用于识别辐射后细胞存活的探索性研究。

Raman Spectroscopy: An Exploratory Study to Identify Post-Radiation Cell Survival.

机构信息

Advanced Center for Treatment, Research and Education in Cancer (ACTREC), Tata Memorial Center (TMC), Navi Mumbai, India.

Homi Bhabha National Institute, Mumbai, India.

出版信息

Appl Spectrosc. 2020 May;74(5):553-562. doi: 10.1177/0003702820908352.

Abstract

Resistance to radiotherapy has been an impediment in the treatment of cancer, and the inability to detect it at an early stage further exacerbates the prognosis. We have assessed the feasibility of Raman spectroscopy as a rapid assay for predicting radiosensitivity of cancer cells in comparison to the conventional biological assays. Cell lines derived from breast adenocarcinoma (MCF7), gingivobuccal squamous cell carcinoma (ITOC-03), and human embryonic kidney (HEK293) were subjected to varying doses of ionizing radiation. Cell viability of irradiated cells was assessed at different time points using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and Raman spectroscopy, and colony-forming capability was evaluated by clonogenic assay. Radiosensitivity observed using MTT assay was limited by the finding of similar cell viability in all the three cell lines 24 h post-irradiation. However, cell survival assessed using clonogenic assay and principal component linear discriminant analysis (PC-LDA) classification of Raman spectra showed correlating patterns. Irradiated cells showed loss of nucleic acid features and enhancement of 750 cm peak probably attributing to resonance Raman band of cytochromes in all three cell lines. PC-LDA analysis affirmed MCF7 to be a radioresistant cell line as compared to ITOC-03 and HEK293 to be the most radiosensitive cell line. Raman spectroscopy is shown to be a rapid and alternative assay for identification of radiosensitivity as compared to the gold standard clonogenic assay.

摘要

放射抗拒一直是癌症治疗的障碍,而早期无法检测到这种情况则进一步恶化了预后。我们评估了拉曼光谱作为一种快速检测方法的可行性,以预测癌症细胞对放射敏感性,与传统的生物学检测方法相比。我们将源自乳腺腺癌(MCF7)、牙龈颊鳞状细胞癌(ITOC-03)和人胚肾(HEK293)的细胞系暴露于不同剂量的电离辐射中。在不同的时间点,使用 MTT(3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化物)测定法和拉曼光谱评估照射细胞的细胞活力,通过集落形成能力评估细胞的克隆形成能力。MTT 测定法观察到的放射敏感性受到限制,因为在照射后 24 小时,所有三种细胞系的细胞活力都相似。然而,使用集落形成测定法和拉曼光谱的主成分线性判别分析(PC-LDA)分类评估的细胞存活情况显示出相关模式。照射后的细胞显示出核酸特征的丧失和 750cm峰的增强,这可能归因于所有三种细胞系中细胞色素的共振拉曼带。PC-LDA 分析证实 MCF7 是一种放射抗性细胞系,而 ITOC-03 和 HEK293 则是最敏感的细胞系。与金标准集落形成测定法相比,拉曼光谱被证明是一种快速的替代检测方法,用于鉴定放射敏感性。

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