Effects of Dichloromethane and N-Butanol Fractions of Nigella sativa on ACHN and GP-293 Cell Line Morphology, Viability, and Apoptosis.

Samira Shahraki, Sara Hosseinian, Elham Shahraki, Mehdi Kheirandish, Abolfazl Khajavirad
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Abstract

Background: Renal cell carcinoma (RCC) is among the top death-causing cancers. Medicinal herbs can also have beneficial effects on RCC treatment. In this project, we aimed to study the antitumor effect of dichloromethane and N-butanol fractions of hydroalcoholic extract of Nigella sativa (N. sativa) on the morphology, viability, and apoptosis of ACHN (human renal adenocarcinoma) and GP-293 (normal renal epithelial) cell lines.

Materials and methods: In this experimental study, N-butanol and dichloromethane fractions of N. sativa were obtained, and ACHN and GP293 cell lines were treated with various concentrations of dichloromethane (0-100 μg/mL) and N-butanol (0-12.5 μg/mL) fractions for 24, 48, and 72 hours. Then, morphological changes, viability, and apoptosis were investigated.

Results: Our results indicated that dichloromethane and N-butanol fractions cause morphological changes and significant decreases in the percentage of live cells in the ACHN cell line, in a dose- and time-dependent manner. In the GP-293 cell line, however, a lower toxicity was observed in comparison with that found for ACHN. The results of flow cytometry showed an apoptotic effect of dichloromethane and N-butanol fractions on the ACHN cell line but a higher rate of apoptosis induction for the total extract compared to the two fractions in the renal cancer cell line compared to the normal cell line.

Conclusion: Our findings demonstrated that these two fractions of N. sativa induce inhibitory effects on the ACHN cell line morphology and viability. These effects were lower than those induced by the total extract. In addition, the two fractions caused more marked effects in the renal cancer cell line compared with the GP-293 cell line.

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二氯甲烷和正丁醇组分对黑草ACHN和GP-293细胞株形态、活力和凋亡的影响
背景:肾细胞癌(RCC)是导致死亡的主要癌症之一。草药对肾小球癌的治疗也有有益的作用。本课题旨在研究黑草(Nigella sativa, N. sativa)水醇提取物二氯甲烷和正丁醇组分对人肾腺癌(ACHN)和正常肾上皮细胞(pg -293)形态、活力和凋亡的影响。材料与方法:本实验提取芥蓝的正丁醇和二氯甲烷组分,分别用不同浓度的二氯甲烷(0 ~ 100 μg/mL)和正丁醇(0 ~ 12.5 μg/mL)处理ACHN和GP293细胞株24、48和72 h。然后观察细胞形态学变化、细胞活力和细胞凋亡。结果:我们的研究结果表明,二氯甲烷和正丁醇组分引起ACHN细胞系的形态改变和活细胞百分比的显著降低,并呈剂量和时间依赖性。然而,在GP-293细胞系中,与ACHN相比,观察到的毒性较低。流式细胞术结果显示,二氯甲烷和正丁醇组分对肾ACHN细胞株有凋亡作用,但总提取物对肾ACHN细胞株的凋亡诱导率高于正常细胞。结论:本研究结果表明,这两个部位对ACHN细胞系形态和活力具有抑制作用。这些作用低于总提取物诱导的作用。此外,与GP-293细胞株相比,两组分对肾癌细胞株的作用更为显著。
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