Patel Apurva, Soni Anjali, Siddiqi Nikhat J, Sharma Preeti
1Department of Biotechnology, Veer Narmad South Gujarat University, Surat, Gujarat 395007 India.
2Department of Biochemistry, College of Science, King Saud University, Riyadh, 11495 Saudi Arabia.
3 Biotech. 2019 Feb;9(2):58. doi: 10.1007/s13205-019-1585-z. Epub 2019 Jan 29.
(TT), a herb belonging to Zygophyllaceae family is widely used due to its medicinal properties. This study was undertaken to elucidate the anticancer mechanism of TT on MCF-7 breast cancer cells. Cytotoxic effect of the herb was assessed by 3-(4,5-diethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Apoptotic potential was assessed through DNA fragmentation, TUNEL and caspase 3 activity assays. Expressions of genes regulating the apoptotic pathway were examined by RT-PCR and expression of proteins was analyzed by immunocytochemistry. The result of MTT assay revealed that methanolic and saponin extracts from leaves and seeds of TT were cytotoxic to MCF-7 cells. Cytotoxicity studies on peripheral blood mononuclear cells (PBMC) proved that TT extracts were non-toxic to non-malignant cells. Treatment of human breast cancer MCF-7 cells with seed and leaf methanol and saponin extracts of TT resulted in fragmentation of DNA and induction of apoptosis. This was evident by agarose gel electrophoresis of DNA and TUNNEL assay. The extracts of TT also caused a significant increase in caspase 3 activity in MCF-7 cells. TT extracts caused an induction of intrinsic apoptotic pathway which was evident by the upregulation in the expression of Bax and p53 genes and downregulation in the expression of Bcl-2. FADD, AIF and caspase 8 genes were also upregulated indicating the possible induction of extrinsic apoptotic pathway. Therefore, our results suggest that the (TT) extracts may exert their anticancer activity by both extrinsic and intrinsic apoptotic pathways.
蒺藜（TT）是一种蒺藜科草本植物，因其药用特性而被广泛使用。本研究旨在阐明TT对MCF - 7乳腺癌细胞的抗癌机制。通过3 -（4,5 - 二甲基噻唑 - 2 - 基）- 2,5 - 二苯基四氮唑溴盐（MTT）法评估该草药的细胞毒性作用。通过DNA片段化、TUNEL和半胱天冬酶3活性测定评估凋亡潜力。通过RT - PCR检测调节凋亡途径的基因表达，并通过免疫细胞化学分析蛋白质表达。MTT分析结果显示，TT叶和种子的甲醇提取物和皂苷提取物对MCF - 7细胞具有细胞毒性。对外周血单核细胞（PBMC）的细胞毒性研究证明，TT提取物对非恶性细胞无毒。用TT的种子和叶甲醇提取物及皂苷提取物处理人乳腺癌MCF - 7细胞导致DNA片段化并诱导凋亡。DNA琼脂糖凝胶电泳和TUNEL测定证明了这一点。TT提取物还导致MCF - 7细胞中半胱天冬酶3活性显著增加。TT提取物诱导了内源性凋亡途径，这通过Bax和p53基因表达上调以及Bcl - 2表达下调得以体现。FADD、AIF和半胱天冬酶8基因也上调，表明可能诱导了外源性凋亡途径。因此，我们的结果表明，TT提取物可能通过外源性和内源性凋亡途径发挥其抗癌活性。
