Department of Histology and Embryology, School of Medicine, Süleyman Demirel Universtiy, Isparta, Turkey.
Department of Medical Biology, School of Medicine, Süleyman Demirel Universtiy, Isparta, Turkey.
J Food Biochem. 2021 May;45(5):e13719. doi: 10.1111/jfbc.13719. Epub 2021 Mar 29.
Apoptotic effects of secoisolariciresinol diglucoside (SDG) in 2D and 3D cultures of SW480 cells were investigated. 40-200 μM SDG was used and IC50 values were determined for three different time intervals as 24, 48, or 72 hr for further experiments. BrdU, TUNEL, AIF, and caspase-3 stainings were used. SDG inhibited cell proliferation almost half and half for all time intervals in 2D and 3D cultures and also, induced apoptosis. Apoptotic cell percentages in the control group for 24, 48, and 72 hr were 27.00%, 29.00%, and 28.00%, respectively, while in the SDG treatment group were 59.00%, 61.00%, and 62.00%, respectively. In the spheroid cell culture, apoptotic cell percentages in the control group for 24, 48, and 72 hr were 6.90%, 7.20%, and 7.10%, respectively, while in the SDG treatment group were 19.50%, 19.50%, and 20.70%, respectively. Caspase-3 and AIF antibodies were used to indicate caspase-dependent and -independent apoptotic pathways. Significant increases were seen in both AIF and caspase-3 stainings when compared to the control group but caspase-3 staining results were significantly greater when compared to the AIF staining at all time intervals (p < .05). To prove this, CASP3 gene expression was evaluated by RT-qPCR. Unlike staining results, there was no statistically significant change at 24 hr in 2D and 3D cultures. But, significant upregulation at 48 (2.32-fold in 2D and 2.46-fold in 3D) and 72 hr (5.04-fold in 2D and 6.45-fold in 3D) were seen. PRACTICAL APPLICATIONS: Colon cancer is one of the most prevalent cancer in the developed countries and its etiology is complex. Although the underlying mechanisms are mostly unknown, the link between diet and colon cancer is known and dietary habits can promote cancer or protect against it. In recent years, flaxseed is accepted as a significant functional food ingredient and feeding with it could help in to prevent cancer. Secoisolariciresinol diglucoside is a flaxseed lignan and is metabolized to mammalian lignans by the gut. In the present study, SDG was evaluated for its apoptotic effects in colon carcinoma cell line via monolayer and spheroid cultures using immunohistochemical and gene expression techniques. Findings of this study suggest that SDG may protect against cancers and in particularly against colon cancer and further investigations has to be carried out for detailed underlying mechanisms.
研究了芝麻素二葡萄糖苷 (SDG) 在 SW480 细胞的 2D 和 3D 培养物中的凋亡作用。使用 40-200 μM 的 SDG，并确定了三种不同时间间隔（24、48 或 72 小时）的 IC50 值，以进行进一步的实验。进行了 BrdU、TUNEL、AIF 和 caspase-3 染色。SDG 几乎对半抑制了 2D 和 3D 培养物中所有时间间隔的细胞增殖，并且还诱导了细胞凋亡。对照组中 24、48 和 72 小时的凋亡细胞百分比分别为 27.00%、29.00%和 28.00%，而 SDG 处理组中分别为 59.00%、61.00%和 62.00%。在球体细胞培养物中，对照组中 24、48 和 72 小时的凋亡细胞百分比分别为 6.90%、7.20%和 7.10%，而 SDG 处理组中分别为 19.50%、19.50%和 20.70%。使用 caspase-3 和 AIF 抗体来指示 caspase 依赖性和非依赖性凋亡途径。与对照组相比，AIF 和 caspase-3 染色均显着增加，但在所有时间间隔内，caspase-3 染色结果均显着大于 AIF 染色（p <.05）。为了证明这一点，通过 RT-qPCR 评估了 CASP3 基因表达。与染色结果不同，2D 和 3D 培养物在 24 小时时没有统计学上的显着变化。但是，在 48 小时（2D 中为 2.32 倍，3D 中为 2.46 倍）和 72 小时（2D 中为 5.04 倍，3D 中为 6.45 倍）观察到显着的上调。实际应用：结肠癌是发达国家最常见的癌症之一，其病因复杂。尽管其潜在机制大多未知，但饮食与结肠癌之间的联系是已知的，饮食习惯可以促进癌症或预防癌症。近年来，亚麻籽被认为是一种重要的功能性食品成分，食用它有助于预防癌症。芝麻素二葡萄糖苷是亚麻籽木脂素，通过肠道代谢为哺乳动物木脂素。在本研究中，通过单层和球体培养物，使用免疫组织化学和基因表达技术，评估 SDG 在结肠癌细胞系中的凋亡作用。这项研究的结果表明，SDG 可能有助于预防癌症，特别是预防结肠癌，需要进一步研究以了解其潜在的机制。
