Chemopreventive Activity of Ferulago angulate against Breast Tumor in Rats and the Apoptotic Effect of Polycerasoidin in MCF7 Cells: A Bioassay-Guided Approach

Abstract

Ferulago angulata leaf hexane extract (FALHE) was found to be a potent inducer of MCF7 cell apoptosis. The aims of the present study were to investigate the in vivo chemopreventive effect of FALHE in rats, to identify the contributing anticancer compound in FALHE and to determine its potential mechanism of action against MCF7 cells. Thirty rats harboring LA7-induced breast tumors were divided into five groups: tumor control, low-dose FALHE, high-dose FALHE, treatment control (tamoxifen) and normal control. Breast tissues were then subjected to histopathological and immunohistochemical analyses. A bioassay-guided investigation on FALHE was performed to identify the cytotoxic compound and its mechanism of action through flow cytometry, real-time qPCR and western blotting analyses. An in vivo study showed that FALHE suppressed the expression of the tumor markers PCNA and Ki67. The tumor size was reduced from 2031 ± 281 mm3 to 432 ± 201 mm3 after FALHE treatment. FALHE administration induced apoptosis in breast tumor cells, and this was confirmed by high expression levels of Bax, p53 and caspase 3. Cell cycle arrest was suggested by the expression of p21 and p27. The in vitro experimental results resulted in the isolation of polycerasoidin as a bioactive ingredient of FALHE with an IC50 value of 3.16 ± 0.31 μg/ml against MCF7 cells. Polycerasoidin induced mitochondrial-dependent apoptosis in breast cancer cells via caspase activation and changes in the mRNA and protein expression of Bax and Bcl-2. In addition, flow cytometric analysis demonstrated that the treated MCF7 cells were arrested at the G1 phase, and this was associated with the up-regulation of p21 and p27 at both the mRNA and protein levels. The results of the present study reinforce further investigations scrutinizing the promising potential of the F. angulata chemical constituents as breast cancer chemopreventive agents.

Chemoprevention of LA7-Induced Mammary Tumor Growth by SM6Met, a Well-Characterized Cyclopia Extract

“…Of the SOC therapies investigated, TAM (10 mg/kg BW), a SERM, significantly ( P < 0.01) decreased tumor volume progression and a substantial reduction in tumor volume was observed until day 14. Interestingly, TAM is the only SOC endocrine therapy that has previously been investigated on LA7-induced tumor growth and the inhibitory effect of TAM observed in the current study is in line with results of previous investigators ( Shamsabadi et al, 2013 ; Karimian et al, 2015 ). This is the first study to report on the effects of the SOC therapies, FUL and LET, on LA7 mammary tumorigenesis ( Figure 2B and Supplementary Figure S2 ).…”

“…The advantages of this model are the strong tumorigenic properties of the cells and a relatively short duration, within 7–10 days, of tumor development. This is a significantly shortened experimental protocol compared to the 8–10 weeks of tumor latency in chemically induced [methyl- N -nitrosourea (MNU) and DMBA] mammary carcinogenic models ( Abbasalipourkabir et al, 2010 ; Karimian et al, 2015 ; Alvarado et al, 2017 ). Not only is the LA7 model a faster method of tumor induction, but it is also ideally suited for testing therapies specifically targeting late stage carcinogenesis (progression) while circumventing the earlier phases.…”

Variation of Chemical Constituents and Antiradical Capacity of Nine Ferulago angulata Populations from Iran

“…For instance, different extracts and coumarins of the plant exerted moderate antiradical scavenging, antibacterial, and antioxidant activities . Moreover, in the literature antioxidant activity, in vivo anxiolytic and antidepressant for the EO, along with cytotoxicity, apoptosis‐inducing, and in vitro anticancer effects of the extracts were reported. Furthermore, the AcOEt and methanolic extract of F. angulata previously revealed the highest levels of phenolic compounds among its various extracts…”

<em>Kelussia odoratissima</em> Mozaff. activates intrinsic pathway of apoptosis in breast cancer cells associated with S phase cell cycle arrest via involvement of p21/p27 in vitro and in vivo

“…After the experiments, an intraperitoneal injection mixture of xylazine (10 mg/kg body weight) and ketamine HCl (150 mg/kg body weight) was used to anesthetize the animals. 19 , 20 …”

“…The tumor tissue sections were subjected to a DeadEnd™ Fluorometric TUNEL system (Promega Corporation, Madison, WI, USA), according to the manufacturer’s instructions, and then captured using a confocal microscope (Zeiss, LSM 70) using standard fluorescein filters. 19 …”