诱导植物愈伤组织中迷迭香酸生物合成基因及其对乳腺癌细胞系的抗增殖活性。

Induction of Biosynthetic Genes Related to Rosmarinic Acid in Plant Callus Culture and Antiproliferative Activity Against Breast Cancer Cell Line.

出版信息

Pak J Biol Sci. 2020 Jan;23(8):1025-1036. doi: 10.3923/pjbs.2020.1025.1036.

DOI:10.3923/pjbs.2020.1025.1036

Abstract

BACKGROUND AND OBJECTIVE

Rosmarinic acid is considered as one of the most important secondary metabolites in medicinal plants especially of family Lamiaceae. Rosmarinic acid can prevent both the tumor initiation and promotion stages of carcinogenesis. The aim of current study was to evaluate the antiproliferative effects of Hyssopus officinalis and Thymus vulgaris callus crude extracts contained rosmarinic acid on breast cancer cells with correlation to phenylpropanoid biosynthetic pathway genes expression.

MATERIALS AND METHODS

Calli of both plants were maintained on Murashige and Skoog medium supplemented with kinetin and 2,4-D. Rosmarinic acid was determined spectrophotometrically in both seed-germinated plants (control) and callus tissues. Transcriptional profiling of rosmarinic acid pathway genes was performed with RT-PCR system. The human breast cancer cell line MCF-7 was treated with different levels of crude extracts at different time intervals in order to show their effects on the cell proliferation using a cell viability colorimetric assay (MTT).

RESULTS

The results showed a significant increase of rosmarinic acid content up to 6.5% in callus compared to control. The transcriptional profile of the selected rosmarinic acid genes in callus tissues indicated significant effects on the rosmarinic acid content in both genotypes. T. vulgaris (90 μg mL-1) and H. officinalis (150 μg mL-1) callus extracts had exhibited highest reduction in the cell MCF-7 viability after 48 h of exposure.

CONCLUSION

It was concluded that rosmarinic acid production increased in callus tissue, showed the higher gene expression levels and remarkably inhibited growth of human breast cancer cell line.

摘要

背景与目的

迷迭香酸被认为是药用植物中最重要的次生代谢物之一，尤其是唇形科植物。迷迭香酸可以预防肿瘤的发生和促进阶段。本研究的目的是评估含有迷迭香酸的海索草和普通百里香愈伤组织粗提取物对乳腺癌细胞的抗增殖作用，并与苯丙素生物合成途径基因表达相关联。

材料与方法

将两种植物的愈伤组织在添加激动素和 2,4-D 的 Murashige 和 Skoog 培养基上维持。使用分光光度法测定两种植物的种子萌发植株（对照）和愈伤组织中的迷迭香酸含量。使用 RT-PCR 系统进行迷迭香酸途径基因的转录谱分析。为了证明不同浓度的粗提取物在不同时间间隔对 MCF-7 人乳腺癌细胞增殖的影响，采用细胞活力比色法（MTT）进行处理。

结果

结果表明，与对照相比，愈伤组织中的迷迭香酸含量显著增加，达到 6.5%。两种基因型的愈伤组织中选定的迷迭香酸基因的转录谱表明对迷迭香酸含量有显著影响。T. vulgaris（90μgmL-1）和 H. officinalis（150μgmL-1）愈伤组织提取物在暴露 48 小时后对 MCF-7 细胞活力的抑制作用最强。