Anti-cancer property of Lenzites betulina (L) Fr. on cervical cancer cell lines and its anti-tumor effect on HeLa-implanted mice

Abstract

In global scenario cervical cancer is increasing. New drugs from natural compounds are in search. Mushrooms are now recognized as miniature pharmaceutical factories producing hundreds of novel constituents. We have taken ethanolic extract Lenzities betulina (LBE) wild mushroom for evaluation of its as anti-cancer property against cervical cancer cell lines e.g. HeLa, CaSki and SiHa and anti tumor activity against HeLa implanted tumor on mice. The extraction was done by dip and stirring method in 90% ethanol for 72 h. For evaluation of anti-cervical cancer, several assays were performed such as MTT assay, cell morphology by phase contrast microscope and F-action polymerization by Laser scanning confocal microscope and nuclear morphology DAPI staining under inverted fluorescence microscope, MMP, ROS, cell cycle, autophagy and stem cell population by flow cytometry and DNA laddering were done. Western blotting was done for protein expression. To evaluate anti-metastatic activity, anti-cologenic assay and wound healing assay were adopted. For chemo-analysis of the LBE, GC-MS was done. The results from Cytotoxicity assay showed that at highest dose of LBE (1000 µg/ml) after 24 h, percentage of cell inhibitions were 85.13 %, 77.13 % and 47.70 % against HeLa , CaSki and SiHa respectively and the calculated IC 50 values were 492.52 ± 2.6 µg/ml, 612.22 ± 4.2 µg/ml, and 1210.30 ± 6.4 µg/ml respectively . Depending upon the cytotoxicity screening, HeLa cell line was considered for the further studies. Cell morphology study exhibited that LBE treated HeLa cells became round from normal spindle shape. DAPI staining showed that LBE treated nucleus became condensed and fragmented. DNA fragmentation at 230 and 300 base pair zone from agarose gel assay was observed. LBE induced ROS generation and reduced MMP. It up regulated the expression of apoptotic genes and p53 while down regulated Bcl2, pro-caspase 3 and pro caspase-9 gene. Cell cycle was arrested at G2/M checkpoint. Autophagic induction was exhibited by vacuole formation in treated cells. CSC population of treated cells was reduced and F-actin polymerization was observed in treated cells. In addition, LBE suppressed metastatic nature by inhibition of cell migration and colonization. The inhibition of growth of the tumors in HeLa cell-implanted mice showed that treatment with 50 mg LBE/kg of body weight of mice led to a marked reduction in the volume (93.22 ± 9.2 %) and weight (90.42 ±9.55 %) of the tumors. The GC-MS profile of LBE shows that out of 69 compounds, 9, 12-Octadecadienoic acid (Z, Z) and 8, (3.beta22E) are in a significantly higher proportion with the percentage peak area 22.13 and 19.72 respectively. Library search for bioactivity showed that these compounds are anti-cancerous and interestingly 4'-Hydroxy-6methoxyaurone binding with P-glycoprotein inhibits the cancer cells to become drug resistant. In conclusion, LBE is very prominent anti-cervical cancer having a lot of anti-cancerous compounds which are probably acting synergistically. This report of anti-ce...

Biodiversity and Bioprospecting of Fungal Endophytes from the Antarctic Plant Colobanthus quitensis

“…Interestingly, other authors demonstrated that, by using the MTT assay, the ethanol extract of Lenzites betulina strongly reduced the viability of several types of cancer cells, and this effect appeared to be induced through ROS generation, reduction of the matrix metalloproteinases, P53 up-regulation, and Bcl2, pro-caspase-3, pro-caspase-9 and proline-rich acidic protein down-regulation [82]. The cytotoxic effect of our extracts appeared to be more intense than that of extracts derived from non-Antarctic species, although tested on different cell lines [82]. Interestingly, a positive correlation between TPC and the ability to inhibit cell growth was observed for all extracts regardless of the types of used cells.…”

Functional activities of beta-glucans in the prevention or treatment of cervical cancer

“…Some other studies have demonstrated that also lentinan, a lentinula edodes-derived beta-glucan, can improve the efficacy of chemo and radiotherapy in many cancers including cervical cancer [158]. Ghosh et al [136] also worked on effects of an ethanolic extract from Lenzities betulina named LBE on cervical cancer cell lines and observed anti-cancer effects against HeLa, CaSki, and SiHa.…”