Lemon Balm Extracts Prevent Breast Cancer Progression In Vitro and In Ovo on Chorioallantoic Membrane Assay

Abstract

Breast cancer is the most frequently diagnosed malignant pathology, representing the primary cause of cancer death in women. Natural products are an appealing strategy to limit the progression of the disease. Targeting angiogenesis in breast cancer may positively impact on poor prognosis of breast cancer. As source of natural compounds, we investigated the leaves of Melissa officinalis L. (MO), known as lemon balm, an aromatic plant that spontaneously grows in the South and Western areas of Romania, being traditionally recommended as anxiolytic, antispasmodic, or as digestive remedy. Our aim was to investigate the phytochemical profiling and the antiangiogenic and chemopreventive bioactivity of MO from Banat region, on breast cancer. Two ethanolic extracts of MO (MOE96 and MOE70) and one methanolic extract (MOM80) were subjected to polyphenol and triterpene profiling by HPLC-MS, and the antioxidant capacity was evaluated. The antiangiogenic potential was investigated using the chorioallantoic membrane assay (CAM). The MTT(3-(4,5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide) assay was used to investigate the cytotoxic effects on MCF-7 and MDA-MB-231breast cancer cells, as well as on MCF-10A normal breast epithelial cells, while apoptosis was performed by DAPI staining. Rosmarinic acid (RA) and ursolic acid (UA) were revealed as dominant phytocompounds. The highest concentration in phytochemicals were found in MOM80; MOE96 was more concentrated in UA, while MOE70 extracted more RA. MOE96 inhibited cancer progression and angiogenesis in the in ovo CAM model using MDA-MB-231 cells, inhibiting breast cancer progression and angiogenesis for the MDA-MB-231 breast cancer cell line; no secondary tumoral areas were registered, indicative for a preventive effect against breast tumor cell invasiveness. The highest cell inhibitory activity was also exhibited by MOE96, in particular against the estrogen receptor positive MCF7 breast cancer cell line, with no cytotoxic effect on healthy cells. The estrogen receptor positive MCF7 cell line proved to be more sensitive to the extract antiproliferative activity than the triple negative MDA-MB-231 breast cancer cell line. Nevertheless, the chemopreventive potential of MOE96 extract is phenotype-dependent and is rather related to the apoptosis and antiangiogenic effects suggesting a multitargeted mechanism of action due to its multiple compound composition next to a concentration ratio of RA : UA in favor of UA.

Comparative Toxicological In Vitro and In Ovo Screening of Different Orthodontic Implants Currently Used in Dentistry

“…On the third day of incubation, a volume of 5–7 mL of albumen was extracted to allow the chorioallantoic membrane to detach from the eggshell so that the blood vessels could be more easily observed. On the fourth day of incubation, a window was cut in the upper part of the egg, which was then covered with adhesive tape, and the eggs were reintroduced into the incubator until the day the experiment began [ 28 , 29 ].…”

Melissa officinalis L. Aqueous Extract Exerts Antioxidant and Antiangiogenic Effects and Improves Physiological Skin Parameters

“…The supernatant was collected and submitted to LC-MS analysis. All polyphenols were separated on a reverse phase Zorbax Eclipse Plus C18 column (3.0 × 100 mm × 3.5 microns) by gradient elution with a mobile phase consisting of a mixture of 0.1% acetic acid and methanol as follows: 5% methanol up to 5 min, 42% methanol for 38 min, the same proportion up to 41 min and 5% methanol for 1 min as described before [16]. The LC-MS parameters were the following: injection volume, 10 µL; flow rate, 1 mL/min; column temperature, 40 • C; UV detection at 330 and 370 nm; MS detection by electrospray ionization (ESI) in the single ion monitoring mode (SIM); capillary voltage, 3500 V; dry gas flow, 12 L/min at 350 • C; nebulizer pressure, 55 psig; fragmentor 70.…”

“…People frequently consume Melissa officinalis herbal tea to ensure a good digestion, to reduce gastrointestinal disorders, to avoid sleep disturbance and for its antispasmodic properties [9,10]. Furthermore, many research studies have shown that MO extract possesses promising antitumor potential in various human cancer cell lines [11][12][13][14][15][16]. Cancer, a highly aggressive and heterogenic disease, is considered one of the causes for the elevated mortality rates worldwide [17].…”

An In Vitro Evaluation of the Molecular Mechanisms of Action of Medical Plants from the Lamiaceae Family as Effective Sources of Active Compounds against Human Cancer Cell Lines

“…The critical moment is tumor-induced angiogenesis. Melissa officinalis leaf extract halted neovascularization in breast adenocarcinoma MDA-MB-231 cells [ 202 ]. Antiangiogenic effects were also exerted by Prunella vulgaris root extract in MCF-5 cells [ 150 ].…”