Abstract
Objective:In this study, we aimed to harness some solvent extracts of one wild mushroom Hexagonia glabra and test their anti-cancer activity against cervical human cell lines, namelyHeLa, SiHa, and CaSki. Methods:It includes cell morphological study by microscope, nuclear morphology by DAPI staining under fluorescence microscopy, apoptosis assay by fluorescence technique, anti-proliferation by MTT assay and expression of apoptotic and anti-apoptotic genes by Western blotting and cell cycle analysis was done. Results:The selected cervical cancer cells were treated separately with 150 µg/mL of three extracts, namely of ethanolic (EE), ethyl acetate (EAE), and water extract (WE) and exhibited features like round, shrink and dead. All extracts caused apoptosis in cell lines and EE had the highest effect in this regard. The percentages of apoptotic cells in HeLa, SiHa and CaSki, at the same concentration of EE were 79.23, 75.42, and 76.36% respectively. Cytotoxicity assay showed that all three extracts (50 – 250 μg/mL) were potent for inhibition of cell growth of three cell lines and again EE had the highest effect. The percentages of cell growth inhibition in HeLa, SiHa, and CaSki cells treated with EE at 24 h at 50 µg/mL were 45.79±4.11, 41.66±4.03, and 36.72±2.67, while they were 74.23±7.45, 62.31±5.97, and 54.23±5.04 at 150 µg/mL concentration. At 250 µg/mL concentration, the percentages of cell growth inhibition were 94.25 ±8.11, 90.02 ±8.67, and 85.43±6.21, respectively. The expression of apoptotic gene (Caspase 3, 9) and tumor guard gene (p53), as their proteins in Western blotting increased . However, anti-apoptotic BcL2 gene of all cell lines was decreased following treatment with extracts. In addition, the cell cycle analysis (CaSki cell) showed that treatment (EE) arrested at G2/M check point cell cycle. Conclusion:All extracts of this mushroom were active in arresting growth of three cell lines and EE had the highest effect, indicating that this mushroom can be a valuable source of anticancer agents.
Highlights
Cancer is one of the most complex killer diseases among human beings
Cell morphological study of all three cell lines under phase contrast microscopy Figure 1 showes that HeLa cells treated with 150 μg/mL of EE resulted in more round, shrunken, and membrane blabbing
We found that treatment with EE, ethyl acetate (EAE), and water extract (WE) at the concentration of 150μg/mL decreased BcL2 gene expression (Figure 4), while increased the expression of apoptosis genes, namely caspase 3 and caspase 9 in all three cell lines in vitro
Summary
Hurdles and hopes for cancer treatment are going on, but scientists search the new natural compound that can be applied as an anticancer remedy (Hunt, 2002). Searching for natural sources to find novel bioactive anticancer compounds may provide generation of drugs (Liu et al, 2010; Lindequist et al, 2010; Xu et al, 2011; Aly et al, 2011). Scientists have paid much attention to mushrooms in cancer management because they have hundreds of novel natural constituents with biological properties with lower toxic side effects and even lower cost (Lee et al, 2014; Shavit et al, 2009). SSG , a homoglucan from Sclerotinia sclerotiorum, triggers the development of TH1 cells via the IL-12 pathway It is isolated from Sparassis crispa and usually enhances the hematopoietic response (Moradali et al, 2007).
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