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褪黑素对人甲状腺未分化癌细胞株毒性和增殖的影响。

Effects of melatonin on the toxicity and proliferation of human anaplastic thyroid cancer cell line.

机构信息

Department of Anatomical Sciences, School of Medicine, Guilan University of Medical Sciences, Rasht, Iran; Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.

Department of Pharmaceutical Biotechnology, School of Pharmacy, Guilan University of Medical Sciences, Rasht, Iran.

出版信息

Acta Histochem. 2021 Apr;123(3):151700. doi: 10.1016/j.acthis.2021.151700. Epub 2021 Mar 2.

Abstract

BACKGROUND

Thyroid carcinoma is the most common endocrine malignancy and anaplastic thyroid carcinoma (ATC) is a rare but most aggressive cancer. Melatonin has enhanced or induced apoptosis in many different cancer cells, however, there has not been any study on the effects of melatonin in the treatment of ATC. In this study, we examined the effect of melatonin on cytotoxicity in the human ATC cell line.

MATERIALS AND METHODS

Cultured ATC cells were treated at melatonin concentrations 0.6, 1, 4, 16, 28 mM for 24 h. The MTT assay was performed to examine cell viability. Cytotoxicity was assayed with the determination of lactic dehydrogenase (LDH) activity. Apoptosis was detected by acridine orange/ethidium bromide and Hoechst 33342 staining. Giemsa staining is considered for evaluating the morphological changes of ATC cells. The reproductive ability of cells to form a colony was evaluated by the clonogenic assay.

RESULTS

Results showed that melatonin could significantly decrease cell viability and the lowest cell viability was observed at 28 mM, 10.26 % ± 0.858 versus control. Similar results were obtained when analyzing LDH activity. The highest LDH levels were observed at 16 and 28 mM (546.08 ± 4.66, 577.82 ± 3.14 munit/mL versus control) that confirmed the occurrence of late apoptosis. The clonogenic assay showed that cells at the high concentration of melatonin (16 and 28 mM) don't enable to form the colony that approved the occurrence of reproductive death.

CONCLUSION

Our results showed a dose-dependent cytotoxic effect of melatonin on ATC cells that significantly decreased cell viability and induced cell reproductive death at the concentration greater than 1 mM and findings suggested that MLT might be useful as an adjuvant in ATC therapy.

摘要

背景

甲状腺癌是最常见的内分泌恶性肿瘤,间变性甲状腺癌(ATC)是一种罕见但最具侵袭性的癌症。褪黑素已增强或诱导许多不同癌细胞凋亡,但尚未有研究探讨褪黑素在 ATC 治疗中的作用。在这项研究中,我们研究了褪黑素对人 ATC 细胞系细胞毒性的影响。

材料和方法

将培养的 ATC 细胞用 0.6、1、4、16、28mM 的褪黑素处理 24 小时。MTT 检测法检测细胞活力。通过测定乳酸脱氢酶(LDH)活性来检测细胞毒性。用吖啶橙/溴化乙锭和 Hoechst 33342 染色检测细胞凋亡。吉姆萨染色用于评估 ATC 细胞的形态变化。用集落形成实验评估细胞的再生能力。

结果

结果表明,褪黑素可显著降低细胞活力,在 28mM 时细胞活力最低,为 10.26%±0.858,与对照组相比。分析 LDH 活性时得到了相似的结果。在 16 和 28mM 时观察到最高的 LDH 水平(546.08±4.66、577.82±3.14 毫单位/毫升,与对照组相比),证实了晚期凋亡的发生。集落形成实验表明,在高浓度褪黑素(16 和 28mM)下的细胞无法形成集落,这证实了生殖死亡的发生。

结论

我们的结果表明褪黑素对 ATC 细胞具有剂量依赖性的细胞毒性作用,显著降低细胞活力,并在浓度大于 1mM 时诱导细胞生殖死亡,这表明 MLT 可能作为 ATC 治疗的辅助药物。

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