Faculty of Life Sciences & Environment, Department of Botany, Goa University, Panjim, India.
Biomarkers. 2020 Dec;25(8):701-710. doi: 10.1080/1354750X.2020.1836025. Epub 2020 Oct 22.
The present study evaluates the anti-tumorigenic potential of leaf methanol extracts of (LMAM).
The cytotoxic activity was assessed in MCF-7 cells by MTT assay at various concentrations ranging from 25-250µg/mL. MCF-7 cells were treated with 50 and 100 µg/mL LMAM for 24 h. To detect LMAM-induced apoptosis; Hoescht 33342 staining along with Cell cycle analysis, Annexin-PI probe as well as oxidative stress damage by reactive oxygen species (ROS) measurements were determined using flow cytometric analysis. While caspase-3 expression levels were studied employing the qRT-PCR method.
LMAM exhibited significant inhibition of MCF-7 cells with an IC value of 85.55 µg/mL. Hoescht staining showed marked morphological features characteristic of apoptosis in LMAM treated cells. Cell cycle analysis confirmed the proven capability of LMAM showing a 30% rise in G phase upon treatment with 100 µg/mL LMAM, thus inducing cell cycle arrest at G phase and a rise in sub G-G population paralleled with a decrease in S phase. Flow cytometric analysis with Annexin V-FITC-PI staining indicated an increase in the early and late apoptotic population with a 3.38% and 19.47% rise respectively when treated with 100 µg/mL LMAM. Treatment with 100 µg/mL LMAM caused an increase in intracellular ROS with MFI value 3334.08. Upregulation of caspase-3 was observed with a 2.18 and 32.47 fold increase compared to control in MCF-7 cells cultured at 50 µg/mL and 100 µg/mL LMAM respectively suggesting caspase-dependent apoptosis.
LMAM proved as a potent ethno-chemopreventive agent and a potential lead in cancer treatment attributable to the synergistic interactive properties of phytoconstituents.
本研究评估了（LMAM）叶甲醇提取物的抗肿瘤潜力。
通过 MTT assay 在 25-250μg/ml 不同浓度下评估 MCF-7 细胞的细胞毒性活性。用 50 和 100μg/ml 的 LMAM 处理 MCF-7 细胞 24 小时。为了检测 LMAM 诱导的细胞凋亡，通过流式细胞术分析，用 Hoechst 33342 染色以及细胞周期分析、 Annexin-PI 探针和活性氧（ROS）测量来确定氧化应激损伤。同时通过 qRT-PCR 方法研究了 caspase-3 的表达水平。
LMAM 对 MCF-7 细胞表现出显著的抑制作用，IC 值为 85.55μg/ml。Hoechst 染色显示，在 LMAM 处理的细胞中出现了明显的凋亡特征形态。细胞周期分析证实了 LMAM 的证明能力，用 100μg/ml 的 LMAM 处理后，G 期细胞上升了 30%，从而诱导细胞周期停滞在 G 期，亚 G-G 群体增加，S 期减少。用 Annexin V-FITC-PI 染色的流式细胞术分析表明，用 100μg/ml 的 LMAM 处理后，早期和晚期凋亡群体分别增加了 3.38%和 19.47%。用 100μg/ml 的 LMAM 处理导致细胞内 ROS 增加，MFI 值为 3334.08。与对照组相比，在 MCF-7 细胞中培养时，caspase-3 的表达上调了 2.18 倍和 32.47 倍，分别为 50μg/ml 和 100μg/ml 的 LMAM，提示 caspase 依赖性凋亡。
LMAM 被证明是一种有效的民族化学预防剂，也是癌症治疗的潜在先导化合物，归因于植物成分的协同相互作用特性。
