In vitro evaluation of Annona muricata L. (Soursop) leaf methanol extracts on inhibition of tumorigenicity and metastasis of breast cancer cells

Abstract

Purpose: The present study evaluates the in-vitro anti-tumorigenic potential of leaf methanol extracts of A. muricata (LMAM). Materials and methods: The cytotoxic activity was assessed in MCF-7 cells by MTT assay at various concentrations ranging from 25-250 Mu g/mL. MCF-7 cells were treated with 50 and 100Mu g/mL LMAM for 24h. 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 qRT-PCR method. Results: LMAM exhibited significant inhibition of MCF-7 cells with an IC sub(50) value of 85.55Mu g/mL. Hoescht staining showed marked morphological features characteristic of apoptosis in LMAM treated cells. Cell cycle analysis confirmed proven capability of LMAM showing 30 percent rise in G sub(1) phase upon treatment with 100Mu g/mL LMAM, thus inducing cell cycle arrest at G sub(1) phase and a rise in sub G sub(0)-G sub(1) 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 3.38 percent and 19.47 percent rise respectively when treated with 100Mu g/mL LMAM. Treatment with 100Mu g/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 50Mu g/mL and 100Mu g/mL LMAM respectively suggesting caspase-dependent apoptosis. Conclusion: LMAM proved as a potent ethno-chemopreventive agent and a potential lead in cancer treatment attributable to the synergistic interactive properties of phytoconstituents.