Abstract
Micromeria fruticosa (L.) Druce subsp. serpyllifolia (Lamiaceae) has been used widely in folk medicine to alleviate various ailments such as abdominal pains, diarrhea, colds, eye infections, heart disorders and wounds. A few reports have confirmed different therapeutic potentialities of its extracts, including the anti-inflammatory, gastroprotective, analgesic, antiobesity and antidiabetic activities. This study aimed to investigate the mechanistic pathway of the antiproliferative activity of the ethanolic extract of M. fruticosa on two different cancer cell lines, namely human breast (mammary carcinoma F7 (MCF-7)) and human colorectal (human colon tumor cells (HCT-116)) cell lines. The 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide tetrazolium (MTT) assay, Annexin V-FITC/PI, caspases 8/9 and cell cycle analyses, qRT-PCR and Western blot were used to assess the effect of M. fruticosa on cytotoxicity, apoptosis, cell cycle, cell cycle-related genes and protein expression profiles in MCF-7 and HCT-116. The extract inhibits cell proliferation in a time- and dose-dependent manner. The half-maximal inhibitory concentration (IC50) for both cell lines was found to be 100 μg/mL. Apoptosis induction was confirmed by Annexin V-FITC/PI, that was related to caspases 8 and 9 activities induction. Furthermore, the cell cycle analysis revealed arrest at G2/M phase. The underlying mechanism involved in the G2/M arrest was found to be associated with the downregulation of CDK1, cyclin B1 and survivin that was confirmed by qRT-PCR and Western blotting.
Highlights
Cancer is reported to be the second leading cause of death, where it causes one death in every six deaths worldwide [1]
The results showed that the M. fruticosa extract decreased the cell viability in a dose- and time-dependent manner in both cell lines (Figure 1) with an IC50 of 208.0, 99.17, and 173.8 μg/mL after 24, 48 and 72 h incubation against the MCF-7 cell line, respectively, and an IC50 of 403.8, 98.8 and 106.3 μg/mL for 24, 48 and 72 h incubation for the HCT-116 cell line, respectively
The concentration of the M. fruticosa extract at 100 μg/mL for 48 h was selected for further analysis
Summary
Cancer is reported to be the second leading cause of death, where it causes one death in every six deaths worldwide [1]. Few reports on M. fruticosa have confirmed its multi-therapeutic potentialities, such as anti-inflammatory, gastroprotective [7,8], analgesic [9], antiobesity and antidiabetic activities [4]. The aqueous extract and the volatile oil of M. fruticosa were previously screened for their antitumor activities against human colon tumor cells (HCT-116) and mammary carcinoma F7 (MCF-7), the underlying mechanism was not explored [10]. The aqueous extract of the plant was reported to induce the growth inhibition of glioblastoma multiforme cells through the induction of oxidative stress in brain cancer cells [11]
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