scholarly journals Anti-Cancer Assessment of a Ramie (Boehmeria nivea L. Gaud.) Leaf Extract Using Mcf-7 Cell Line and a Yeast-Based Bioassay

2021 ◽  
Vol 2 (1) ◽  
pp. 1-12
Author(s):  
Asri Peni Wulandari ◽  
Annisa Abdiwijaya Qaromah ◽  
Karen Kezia Lolowang ◽  
Desi Harneti Putri Huspa ◽  
Ade Zuhrotun

Introduction: One strategy for molecular cancer therapy is to know the key mechanism of cytotoxic compounds that can kill cancer cells. Ramie (Boehmeria nivea L. Gaud.) leaves contain active compounds that have important effects on cancer chemoprevention. Objective: To obtain the active fraction of a Ramie leaf extract in inhibiting the proliferation of MCF-7 breast cancer cell lines and to determine the mechanism of apoptosis induction using MCF-7 and Saccharomyces cerevisiae strains 1140, 1353, and 1138. Method: Fractions were prepared using n-hexane, dichloromethane (CH2Cl2), ethyl acetate, and n-butanol as solvents. All fractions were tested qualitatively through phytochemical. The MTT-based cytotoxicity assay used MCF-7 (in vitro) to obtain the IC50 value, whereas the model system that targets the enzymatic (topoisomerase) used a yeast-based bioassay to obtain the IC12 value. Apoptotic induction of the active fraction in MCF-7 was performed using flow cytometry and qPCR (2-ΔΔCt method). Results: The phytochemical analysis indicated that the extract fraction consisted of alkaloids and steroids. The smallest IC50 value was obtained from the CH2Cl2 fraction as 3.79 g/mL, potentially act as an anticancer. A higher percentage of apoptosis than that of necrotizing cells and live cells was observed through flow cytometry. The CH2Cl2 fraction with an IC12 value < 8000 in strains 1140, 1353, and 1138 consistently showed the mechanism of apoptosis induction as topoisomerase I and II inhibitors. Also, another mechanism could be through the intrinsic pathway, indicated by the highest expression level in p53. Conclusions: The CH2Cl2 fraction of Ramie leaves can inhibits the proliferation of MCF-7 cells in the active and strong categories. The CH2Cl2 fraction induces apoptosis by increasing p53 gene expression and inhibiting topoisomerase I and II. Thus, it showed potential as an anticancer drug candidate.

Author(s):  
Zeinab Abedian ◽  
Niloofar Jenabian ◽  
Ali Akbar Moghadamnia ◽  
Ebrahim Zabihi ◽  
Roghayeh Pourbagher ◽  
...  

Objective/ Background: Cancer is still the most common cause of morbidity in world and new powerful anticancer agents without severe side effects from natural sources is important. Methods: The evaluation of cytotoxicity and apoptosis induction was carried out in MCF-7,HeLa and Saos-2 as cancerous cell lines with different histological origin and human fibroblast served as control normal cell. The cells were treated with different concentrations of chitosan and the cytotoxicity was determined using MTT assay after 24, 48 and 72 h .The mode of death was evaluated by flow cytometry . Results: While both types of chitosan showed significant concentration-dependently cytotoxic effects against the three cancerous cell lines, fibroblast cells showed somehow more compatibility with chitosan. On the other hand, there were no significant differences between LMWC and HMWC cytotoxicity in all cell lines. The flow cytometry results showed the apoptosis pattern of death more in Saos-2 and HeLa while necrosis was more observable with MCF7. Also higher viability with both types of chitosan was seen in fibroblast as normal cells Conclusion: Chitosan shows anticancerous effect against 3 cancerous cell lines, while it is compatible with normal diploid fibroblast cells. Furthermore, it seems that the molecular weight of chitosan does not affect its anticancerous property.


Molecules ◽  
2020 ◽  
Vol 25 (18) ◽  
pp. 4332
Author(s):  
Nurul Izzati Zulkifli ◽  
Musthahimah Muhamad ◽  
Nur Nadhirah Mohamad Zain ◽  
Wen-Nee Tan ◽  
Noorfatimah Yahaya ◽  
...  

A bottom-up approach for synthesizing silver nanoparticles (AgNPs-GA) phytomediated by Garcinia atroviridis leaf extract is described. Under optimized conditions, the AgNPs-GA were synthesized at a concentration of 0.1 M silver salt and 10% (w/v) leaf extract, 1:4 mixing ratio of reactants, pH 3, temperature 32 °C and 72 h reaction time. The AgNPs-GA were characterized by various analytical techniques and their size was determined to be 5–30 nm. FTIR spectroscopy indicates the role of phenolic functional groups in the reduction of silver ions into AgNPs-GA and in supporting their subsequent stability. The UV-Visible spectrum showed an absorption peak at 450 nm which reflects the surface plasmon resonance (SPR) of AgNPs-GA and further supports the stability of these biosynthesized nanoparticles. SEM, TEM and XRD diffractogram analyses indicate that AgNPs-GA were spherical and face-centered-cubic in shape. This study also describes the efficacy of biosynthesized AgNPs-GA as anti-proliferative agent against human breast cancer cell lines, MCF-7 and MCF-7/TAMR-1. Our findings indicate that AgNPs-GA possess significant anti-proliferative effects against both the MCF-7 and MCF-7/TAMR-1 cell lines, with inhibitory concentration at 50% (IC50 values) of 2.0 and 34.0 µg/mL, respectively, after 72 h of treatment. An induction of apoptosis was evidenced by flow cytometry using Annexin V-FITC and propidium iodide staining. Therefore, AgNPs-GA exhibited its anti-proliferative activity via apoptosis on MCF-7 and MCF-7/TAMR-1 breast cancer cells in vitro. Taken together, the leaf extract from Garcinia atroviridis was found to be highly capable of producing AgNPs-GA with favourable physicochemical and biological properties.


2009 ◽  
Vol 27 (15_suppl) ◽  
pp. e14637-e14637
Author(s):  
L. Ko ◽  
J. Allalunis-Turner

e14637 Background: DCA is a generic, orally available, small molecule metabolic modulator that has an established history in the treatment of mitochondrial diseases and lactic acidosis. DCA inhibits pyruvate dehydrogenase kinase (PDK), an inhibitor of pyruvate dehydrogenase, a key enzyme in glucose metabolism. DCA preferentially shifts glucose metabolism in cancer cells from glycolysis to glucose oxidation, reversing the unique aerobic glycolysis found in solid tumors. DCA induced apoptosis in cancer cells as evidenced by the efflux of cytochrome c and apoptosis-inducing factor from the mitochondria. Recent studies suggested apoptosis induction by DCA in glioblastoma, endometrial, prostate, and non-small cell lung cancers. In this study we attempt to establish a link between DCA and apoptosis in breast cancer cell lines. Methods: Six breast cancer cell lines were investigated (BT474, MCF-7, MDA-MB231, MDA- MB468, SKBR3, T47D). Quantitative real-time polymerase chain reaction (RT-PCR) was performed using Taqman probes to identify increased DNA templates of PDK isoforms 1–4, using DCA concentrations from 0 to 20mM. Western blots were performed to identify increased expression of PDK isoforms and correlate findings with Taqman. MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)- 2-(4-sulfophenyl)-2H-tetrazolium) assays were performed to measure decreased mitochondrial activity and cytotoxicity. Flow cytometry using annexin V and propidium iodide was performed to measure apoptosis and cell death. Results: RT-PCR showed increased DNA expression of all PDK isoforms in MDA-MB231 cells after DCA treatment. The effect was most pronounced at 10mM concentration. 10mM of DCA also increased DNA expression of all PDK isoforms in MCF-7 cells, and PDK1 in T47D cells. MTS assays showed increased cell kill and decreased mitochondrial activity in all six cell lines, with IC50 ranging between 20mM and 30 mM. Flow cytometry showed increased apoptosis induced by DCA at IC50 for BT474 and MCF-7 cell lines. Conclusions: Apoptosis appears to play a role in the mechanism of DCA in breast cancer, with increased PDK isoform expressions, cytotoxicity and decreased mitochondrial activity. Data from flow cytometry suggested DCA-induced apoptosis in two cell lines. No significant financial relationships to disclose.


2020 ◽  
Vol 20 (6) ◽  
pp. 1448
Author(s):  
Hersa Milawati ◽  
Winda Sukmawati ◽  
Desi Harneti ◽  
Rani Maharani ◽  
Nurlelasari Nurlelasari ◽  
...  

Three aromadendrane-type sesquiterpenoids, spathulenol (1), 4β,10α-dihydroxyaromadendrane (2), and 4α,10α-dihydroxyaromadendrane (3) were isolated from the stem bark of Aglaia harmsiana (Meliaceae). Compound 3 was isolated for the first time from Aglaia genus. The chemical structures of isolated compounds were elucidated by various spectroscopic methods, including one and two-dimensional NMR, as well as mass spectroscopy analysis. These sesquiterpenoids 1-3 were evaluated for their cytotoxic activity against MCF-7 breast cancer cell lines. The IC50 value of compound 1-3 were 31.65 ± 0.1, 8.41 ± 0.04 and 2.80 ± 0.02 µM, respectively. Among the aromadendrane-type sesquiterpenoids, compounds 2 and 3, which do not have a double bond, showed higher activity than compound 1. Whereas, compound 3 showed the strongest activity indicate that α configuration of hydroxyl group increases the cytotoxic activity.


2017 ◽  
Vol 9 (4) ◽  
pp. 597
Author(s):  
Vaishali Rai ◽  
Vinitha Ramanath Pai ◽  
Surya Ram Duwal

<p align="left">The anticancer activity of crude extracts of the leaves of <em>Tridax procumbens, </em>against two breast cancer cell lines-MCF-7 (benign) and MDA-MB-231 (metastatic) were investigated and an attempt was made to identify the anticancer principle. The extracts with methanol (TPM), ethanol (TPE) and chloroform (TPC) as solvents were screened for cytotoxicity by MTT assay against MCF-7 cells. The effective extract was further evaluated on MDA-MB-231 cells. Among the three extracts, TPC was effective at an IC<sub>50</sub> value of 136 µg/ml and 129 µg/ml on MCF-7 and MDA-MB-231 cells respectively. Phytochemical analysis of the extract showed the presence of only steroids and terpenoids and their concentration was high (77.4% w/w). HPLC-MS of the chloroform soluble crude extract revealed a major peak (57.59% concentration) at a retention time of 4.78 min and MS data of this peak revealed presence of two fragments of molecular weight 475.80 and 701.80. The compounds were identified to be 3β, 9β-Dihydroxy-18α-oleanan-28-oic acid, a derivative of oleanolic acid and 3β, 28-Bis (cinnamoyl) betulin respectively, both from the triterpenoid family. </p>


2021 ◽  
Vol 2021 ◽  
pp. 1-16
Author(s):  
Lusia Barek Moses ◽  
Mohd Fadzelly Abu Bakar ◽  
Hasmadi Mamat ◽  
Zaleha Abdul Aziz

The present study was conducted to determine the cytotoxicity effect of Eurycoma longifolia (Jack.) leaf extracts and also its possible anticancer mechanism of action against breast cancer cell lines: non-hormone-dependent MDA-MB-231 and hormone-dependent MCF-7. The leaves of E. longifolia were processed into unfermented and fermented batches before drying using freeze and microwave-oven drying techniques. Obtained extracts were tested for cytotoxicity effect using MTT assay and phenolic determination using HPLC-DAD technique. The most toxic sample was analyzed for its apoptotic cell quantification, cell cycle distribution, and the expression of caspases and apoptotic protein using flow cytometry technique. Fragmentation of DNA was tested using an agarose gel electrophoresis system. The results determined that the unfermented freeze-dried leaf extract was the most toxic towards MDA-MB-231 and MCF-7 cells, in a dose-dependent manner. This extract contains the highest phenolics of gallic acid, chlorogenic acid, ECG, and EGCG. The DNA fragmentation was observed in both cell lines, where cell cycle was arrested at the G2/M phase in MCF-7 cells and S phase in MDA-MB-231 cells. The number of apoptotic cells for MDA-MB-231 was increased when the treatment was prolonged from 24 h to 48 h but slightly decreased at 72 h, whereas apoptosis in MCF-7 cells occurred in a time-dependent manner. There were significant activities of cytochrome c, caspase-3, Bax, and Bcl-2 apoptotic protein in MDA-MB-231 cells, whereas MCF-7 cells showed significant activities for caspase-8, cytochrome c, Bax, p53, and Bcl-2 apoptotic protein. These results indicate the ability of unfermented freeze-dried leaf extract of E. longifolia to induce apoptosis cell death on MDA-MB-231 and MCF-7, as well as real evidence on sample preparation effect towards its cytotoxicity level.


2017 ◽  
Vol 13 (3) ◽  
pp. 249
Author(s):  
Tahani I. Kashar ◽  
Marwa Abdel-Motaal ◽  
Khadija Emran ◽  
Noha A. Sukar

Benzofurane and naphthofuranethiosemicarbazone ligands 2a (L1 )and 2b(L2) and their Cu(II), Co(II), Ni(II), and Pd(II) complexes have been synthesized and characterized by elemental analyses, molar conductance, magnetic susceptibility measurements, IR, NMR, and electronic spectral studies. IR spectra show that the ligands is coordinated to the metal ions in a didentate manner with N,S doner sites. In the presence of tow thiosemicarbazones ligands 2a(L1) and 2b(L2), the corrosion resistance of cast iron in 10.M H2SO4 increases in the order, 2b(L2) >2a(L1). The effect of studied ligands inhibits the corrosion of cast iron extent depending on the structure and high donation of π-electron present inhibitor. Co(II) and Pd (II) complexes possess higher antimicrobial activity than free ligands and other complexes. The [L1Pd Cl2]H2O complexe shows higher IC50 value of 4.6 µg/ml toward human breast cancer cell lines (MCF-7). For other complexes, they are in the range of 2.7– 4.3 µg/ml toward the former cells which is related to their metal complexing ability in enzymes.


Author(s):  
ANNA ANTONY ◽  
NEHA SHRIVASTAV ◽  
SIDDHARTH DUBEY ◽  
ASHUTOSH ACHARYA

Objective: To synthesize silver nanoparticle (AgNP) using the ethanolic leaf extract of Magnolia champaca (MC-AgNP) and to evaluate its cytotoxicity and apoptotic activity in MCF-7 cell line, and to analyse active Caspase-3 expression in inducing apoptosis. Methods: AgNP was green synthesized using ethanolic leaf extract of MC. Shape and size were analysed by SEM and particle size analyser. Cytotoxicity was tested to find IC50 value. Apoptosis was explored by mitochondrial membrane potential (ΔΨm) assay and active Caspase-3 expression study using flow cytometry. Results: AgNP synthesized was observed to be in spherical shape through SEM micrograph with 164.6 nm in size. IC50 value of 15.90 µg/ml was obtained after 24 h of drug treatment. Changes in ΔΨm and positive active Caspase-3 protein expression were observed. Conclusion: The present results suggest that MC-AgNP has potential anticancer property and also induced cell death via apoptosis through caspase dependent pathway in MCF-7 cells.


Sign in / Sign up

Export Citation Format

Share Document