Antiproliferative Activity Of Methanol Extract Research Articles

Phytochemical profile by LC-MS/MS analysis and evaluation of antioxidant, antidiabetic, anti-Alzheimer, and anticancer activity of Onobrychis argyrea leaf extracts

IntroductionStudies on different species of Onobrychis, a member of the Fabaceae family, have revealed a number of biological activities with potential applications in alternative medicine. The phytochemical content of the plant Onobrychis argyrea subsp. argyrea and some of the important biological activities linked to its metabolism are quite limited. Therefore, this study was the first to determine the content of secondary metabolites in this species and to elucidate important biological activities related to metabolism. MethodsLC-MS/MS was used for the quantitative analysis of bioactive compounds in the aerial part of O. argyrea extracts. The antioxidant properties of the extracts were assessed based on their ability to scavenge DPPH radicals and reduce iron ions. The anti-diabetic effect of the extracts was tested by the inhibition of α-amylase and α-glycosidase enzymes, and the anti-Alzheimer's ability was tested by the inhibition of the enzymes AChE and BChE. The cytotoxic effects of the extracts were tested on four different cancer lines, namely A-549, HT-29, MCF-7 and MDA-MB 453 using the XTT assay. The mechanism of the anti-cancer effect was determined by means of flow cytometry analysis on HT-29 cells. ResultsTwenty-nine phytochemicals were identified via the LC-MS/MS analysis in the plant extracts. The most abundant phytochemicals in the extracts were quinic acid, isoquercitrin, epicatechin, and routine, respectively. Antioxidant analysis showed that among all extracts, methanol extract (ME) was most effective in scavenging DPPH (IC50: 23.93 ± 0.96 µg/mL) and in reducing iron ions (53.24 ± 2.14 mgTEAC/g). Enzyme activity studies demonstrated that ME is a potent inhibitor of α-glycosidase (IC50: 15.06±0.64 µg/mL), AChE (IC50: 24.75±0.76 µg/mL), and BChE (IC50: 3.36±0.14 µg/mL). XTT assay results disclosed the strongest anti-proliferative activity of ME against the human colorectal adenocarcinoma HT-29 cell line with an IC50 value of 33.35±0.92 µg/mL. Flow cytometry analysis revealed that ME caused mitochondrial membrane damage in HT-29 cells (**p<0.01) and subsequently induced apoptosis (**p<0.01) by activating the caspase pathway (**p<0.001). ConclusionO. argyrea, rich in bioactive compounds, showed remarkable antioxidant, antidiabetic, Alzheimer's, and anticancer properties in vitro, demonstrating that this plant contains secondary metabolites that may be potential drug candidates for the treatment of metabolic diseases.

LC-MS/MS analysis and diverse biological activities of Hypericum scabrum L.: In vitro and in silico research

This study was designed to determine the phytochemical composition of Hypericum scabrum extracts and investigate their biologically prominent activities. LC-MS/MS analysis revealed the presence of 29 phytochemicals in the plant extracts. The methanol extract (ME) exhibited the strongest DPPH radical scavenging (IC 50 : 18.57 µg/mL) and ferric ion reducing capacity (74.20 mg TEAC/g). Besides, ME displayed the most promising α-glycosidase (IC 50 : 1.88 µg/mL) and acetylcholinesterase AChE (IC 50 : 8.41 µg/mL) inhibitory effect. The XTT assay results displayed the strong anti-proliferative activity of ME against MCF-7 (IC 50 : 56.45 µg/mL) and MDA-MB-453 (IC 50 : 67.79 µg/mL) cell lines. Flow cytometry studies confirmed the mitochondrial-dependent apoptotic effect of ME in MCF-7 cells. Finally, molecular docking studies revealed the best binding affinity of the compounds 1, 9, and 41 with AChE and α-Gly, while 41 exhibited the best affinity against caspase-3, caspase-9 and DNA. To conclude, this study provides essential data for further in vivo studies. • Phytochemical content and biological activities of Hypericum scabrum extracts were enlightened. • Methanol extract (ME) exhibited promising biological activities regarding metabolism and cancer. • •Flow cytometry results revealed the apoptotic effect of ME via mitochondria-dependent pathway. • The major phenolics showed best binding of with AChE, α-Gly, caspase-3, caspase-9 and DNA.

Cytotoxic and antiproliferative potential of methanolic extracts of Asparagus racemosus in MDAMB231 cells

The present study was carried out to evaluate the cytotoxic and antiproliferative activity of methanolic extract of Asparagus racemosus in MDAB-231 cells. The qualitative phytochemical analysis of the extract revealed the presence of steroids, alkaloids, flavonoids, glycosides, saponins and diterpenes. MDAMB-231 cells were maintained in RPMI media containing 10 per cent serum and 1 per cent antibiotic and antimycotic solution. The cells were harvested and seeded to 96 well plate at 5×103 cells/mL, incubated for 24 hours at 37˚C with 5 per cent CO2. The cells were treated with 160, 80, 40, 20 and 10µg/mL of the extract for 24 hours and viability was accessed using MTT Assay. Also cells were plated in 6 well plates at concentrations 3×105 cells/mL and exposed to 80, 40, 20 and 10µg/mL of the extract of Asparagus racemosus for acridine orange ethidium bromide (AO/EB) staining. There was a dose dependent decrease in viability of cells exposed to methanolic extracts of Asparagus racemosus with a viability of 15.55+0.193 per cent for cells treated with 160µg/mL. The IC50 was found to be 91.36+ 0.87 µg/mL. The AO/EB staining revealed that most of the cells were dead in extract treated with 80µg/mL where more than 90 per cent of cells were live when treated with 10µg/mL of the extract.

Evaluation of Cytotoxic Potentials of Microalgae on Hela, Hepg-2, HT-29 and EACC Cells to Develop Anticancer Agents

Finding more effective methods to treat cancer remains a challenge, and development of new therapeutic agents for cancer treatment is essential for continued progress against the disease. This study was conducted to evaluate the in vitro antiproliferative activity of methanolic extracts of Chlorella vulgaris and Nostoc muscorum against Human cervical Cancer cells (HeLa), liver hepatocellular cells (HepG-2), Human colorectal adenocarcinoma cells (HT-29) and Ehrlich Ascites Carcinoma Cells (EACC) in order to determine the bioacitivities of microalgae. Morphological changes of cancer cells treated with the selected extract were observed by phase contrast microscopy. The cells were incubated with increasing concentrations of microalgal extracts for 24, 48, and 72 h, and cell viability was determined by the Sulforhodamine B (SRB) assay. After treatment of cancerous cells with microalgal extract for 48 h, 55% of the cells were viable, 38% were in early apoptosis, 5% were in late apoptosis, and 1% exhibited necrosis. Further studies are needed to discover the mechanisms underlying the antiproliferative activity of microalgal extracts against cancer cell lines and to identify specific bioactive compounds that are responsible for the observed activities

Antiproliferative and antiviral activity of methanolic extracts from Sardinian Maltese Mushroom (Cynomorium coccineum L.)

Cynomorium coccineum is a non-photosynthetic plant that grows in Mediterranean countries and that is amply used in the traditional medicine. The aim of this study was to extend previous studies on the chemical and biological properties of C. coccineum, evaluating the potential antiviral and antiproliferative activity of the methanolic extract. The MTT assay was used for the in vitro cytotoxic studies against human cancer-derived cell lines, while both MTT and plaque reduction (PRT) methods were used to evaluate the potential inhibitory effect of the extract against a panel of mammal viruses. The results obtained showed no selective activity against any DNA and RNA virus but revealed an interesting antiproliferative activity against human leukaemia-derived cell lines.

Inhibitory effects of methanol extracts of selected plants on proliferation of two human melanoma cell lines

Purpose: The aim of the current study was to investigate the in vitro antiproliferative activity of methanolic extracts of six plants regardless of their claimed ethnopharmacological application.Methods: Methanol extracts of different parts of Glycyrrhizaglabra L. (Licorice), Matricaria chamomilla L. (Chamomile), Salvia triloba L. (Sage), Rheum palmatum L. (Rhubarb), Trigonella foenum-graecum L. (Fenugreek) and Sambucusebulus L. (Dwarf Elder) were prepared. The antiproliferative effects of the extracts were tested on two skin cancer melanoma cell lines namely A375.S2 (low tyrosinase expression) and WM 136.1A (high tyrosinase expression) using MTT assay. The IC50 values for the active extracts were determined against the two melanoma cell lines.Results: The methanolic extracts of G.glabra, M. chamomilla, S. triloba, R. palmatum inhibited the melanotic WM1361A proliferation in a dose-dependent manner revealing IC50 values of 35.2, 25.2, 20.6, 17.8, μg/ml, respectively but not A375.S2 cell line. However, the extracts of T. foenum-graecum and S. ebulus did not exhibit any significant cytotoxic activity on both melanoma cell lines.Conclusion: The results of these experiments show that methanol extracts of licorice, chamomile, sage and rhubarb have significant antiproliferative activity onWM1361A cell line; a representative human melanotic melanocyte tumor cell line. This renders these plants as potential sources of new lead compounds for the development of new drugs for melanoma cancer.Keywords: Melanoma, Plant extract, tyrosinase, Licorice, Chamomile, Sage, Rhubarb, WM1361A

HPLC Analysis of Phenolic Compounds from Gypsophila aucheri Boiss. and Investigation of Antioxidant and Cytotoxic Activity of Gypsophila aucheri Boiss. exracts

This study was designed to investigate the antioxidant and antiproliferative activities of Gypsophila aucheri Boiss. extracts as well as phenolic content by RP-HPLC technique. Antioxidant activity potentials of the extracts were evaluated by four different methods namely, 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging capacity tests, cupric ion reducing antioxidant capacity (CUPRAC) method, and metal chelating assay. Antiproliferative activities of the extracts were tested against breast carcinoma (MCF-7), colorectal carcinoma (HT-29), and hepatocellular carcinoma (HepG2) cells. RP-HPLC analysis revealed the presence of eight phenolic compounds in eighteen phenolics scanned. Antioxidant activity results showed that methanol extract of Gypsophila aucheri displayed more pronounced antioxidant activity than water extract according to DPPH and ABTS tests, whereas water extract was superior than methanol extract for CUPRAC and metal chelating activity tests. Cytotoxicity tests demostrated the promising antiproliferative activity of methanol extract against all the tested cancer cell lines with its lower IC50 values. However, both extracts were observed to display moderate biological activity compared to positive controls. The obtained data suggest that Gypsophila aucheri could be evaluated as a promising source for food and nutraceutical industries due to its striking antioxidant and moderate antiproliferative potentials together with high phytochemical profile

Inhibitory effects of methanol extracts of selected plants on the proliferation of two human melanoma cell lines

Purpose: The aim of the current study was to investigate the in vitro antiproliferative activity of methanolic extracts of six plants regardless of their claimed ethnopharmacological application. Methods: Methanol extracts of different parts of Glycyrrhizaglabra L. (Licorice), Matricaria chamomilla L. (Chamomile), Salvia triloba L. (Sage), Rheum palmatum L. (Rhubarb), Trigonella foenum-graecum L. (Fenugreek) and Sambucusebulus L. (Dwarf Elder) were prepared. The antiproliferative effects of the extracts were tested on two skin cancer melanoma cell lines namely A375.S2 (low tyrosinase expression) and WM 136.1A (high tyrosinase expression) using MTT assay. The IC 50 values for the active extracts were determined against the two melanoma cell lines. Results: The methanolic extracts of G.glabra , M. chamomilla , S. triloba , R. palmatum inhibited the melanotic WM1361A proliferation in a dose-dependent manner revealing IC 50 values of 35.2, 25.2, 20.6, 17.8, μg/ml, respectively but not A375.S2 cell line. However, the extracts of T. foenum-graecum and S. ebulus did not exhibit any significant cytotoxic activity on both melanoma cell lines. Conclusion: The results of these experiments show that methanol extracts of licorice, chamomile, sage and rhubarb have significant antiproliferative activity onWM1361A cell line; a representative human melanotic melanocyte tumor cell line. This renders these plants as potential sources of new lead compounds for the development of new drugs for melanoma cancer. Keywords: Melanoma, Plant extract, tyrosinase, Licorice, Chamomile, Sage, Rhubarb, WM1361A

Effect of Methanolic Extract of Dandelion Roots on Cancer Cell Lines and AMP-Activated Protein Kinase Pathway.

Ethnomedicinal knowledge of plant-derived bioactives could help us in discovering new therapeutic compounds of great potential. Certainly, dandelion has been used in traditional ethno-medicinal systems (i.e., Chinese, Arabian, Indian, and Native American) to treat different types of cancer. Though, dandelion is highly vigorous, but the potential mode of action is still unclear. In the current study, the antiproliferative activity of methanolic extracts of dandelion root (MEDr) on cell viability of HepG2, MCF7, HCT116, and normal Hs27 was investigated. It was observed that MEDr (500 μg/mL) drastically decreased the growth of HepG2 cell line, while the effect on MCF7 and HCT116 cell lines was less pronounced and no effect has been observed in Hs27 cell lines. The MEDr also enhanced the phosphorylation level of AMPK of HepG2 cells, which considered crucial in cancer treatment and other metabolic diseases. The AMPK activation by MEDr noticed in the current study has never been reported previously. The results regarding the number of apoptotic cells (HepG2 cells) were in line with the cell viability test. The current observations clearly demonstrated the potency of MEDr against liver cancer with validation that dandelion could control AMPK and thus cancer in the treated cell lines.

COMPREHENSIVE INVITRO EVALUATION OF PHARMACOLOGICAL ACTIVITIES OF SELECTED PLANT EXTRACTS AND GC-MS PROFILING OF FLACOURTIA JANGOMAS FLOWER EXTRACT

Objective: Continued exploration and bio-evaluation of plants hold good as there are an increasing demand and recognition of natural products in disease management. The aim of this study was to investigate the antioxidant, cytotoxic, and antiproliferative activity of methanolic extracts of different parts of six plants. Six plants - Couroupita guianensis, Flacourtia jangomas, Lucuma nervosa, Euphorbia milii, Acalypha hispida, and Hydnocarpus pentandra - were chosen for this study.Methods: The plant parts were extracted with methanol and screened for 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azinobis-(3- ethylbenzothiazoline-6-sulfonic acid (ABTS) radical scavenging abilities. The cytotoxic activity of the extracts was investigated on SCC9 and Calu6 through MTT assay, and cell cycle was analyzed by flow cytometry to determine the antiproliferative activity of the extracts. The flower extract of F. jangomas was further subjected to gas chromatography-mass spectrometry (GC-MS) analysis for purification of the compounds of interest. A twoway ANOVA was done to estimate the effect of the extract between samples remembered at p&lt;0.05 level.Results: Among all the plant extracts, the extract of F. jangomas (flower) showed significant antioxidant potential with IC50 values of 11.16±0.54 μg/mland 12.34±0.37 μg/ml for DPPH and ABTS assays, respectively. GC-MS analysis of the extract revealed the presence of 21 phytocompounds. MTT assay revealed that this extract had promising cytotoxic activity against the two cancer cell lines, Calu6 and SCC9 with IC50 values of 43.57±0.04 μg/ml and 53.42±0.15 μg/ml, respectively. The extract treatment caused significant arrest in G2M phase of cell cycle.Conclusion: F. jangomas flower extract displayed significant antioxidant and antiproliferative activity and can be considered as a potential source ofanticancer compounds.Keywords: Antioxidants, MTT assay, Gas chromatography-mass spectrometry, Calu6, SCC9, Flacourtia jangomas.

Anti-Proliferative and Antimicrobial Activity of Methanolic Extract and SPE Fractions of Artemisia spicigera

Objectives and Methods: This study was conducted to investigate in vitro, methanolic extract and methanol/water fractions obtained from the arial parts of Artemisia spicigera C. Koch (Asteraceae family) and for evaluation of their antiproliferative effects against HT-29, L-929 and A 549 cell lines by MTT assay at different concentrations (1, 10, 100, 1000 µg/mL). Furthermore, this study aimed to detect antimicrobial activities of the mentioned samples: two Gram-positive (Staphylococcus epidermidis and Staphylococcus aureus), two Gram-negative bacteria (Pseudomonas aeruginosa and Escherichia coli) and a fungi (Candida albicanse), using agar well diffusion method. Results: The IC50 values for antiproliferative activity of the methanolic extract, 20%, 40% and 60% SPE (solid phase extraction) fractions, found to be 345.91 ± 28.77, 442.44 ± 83.22, 220.19 ± 43.13 and 579.90 ± 153.19 μg/mL, respectively; in this case, the maximum inhibition percentage belonged to 40% SPE fraction (220.19 ± 43.13 μg/mL). The total methanolic extract of A. spicigera indicated inhibitory activity against Gram- positive strains, S. epidermidis and S. aureus, with MIC values of 150 mg/mL for both bacteria. Among SPE fractions, 20% was the only active one against Gram possitive species with MIC values of 35 mg/mL for both strains. Conclusions: This study revealed that methanolic extract of A. spicigera and its SPE fractions might be regarded as bioactive natural products, which deserves to be further identification and isolation of cytotoxic and antibacterial phytochemicals from them.

Anti-Proliferative and Antimicrobial Activity of Methanolic Extract and SPE Fractions of Artemisia spicigera

Objectives and Methods: This study was conducted to investigate in vitro, methanolic extract and methanol/water fractions obtained from the arial parts of Artemisia spicigera C. Koch (Asteraceae family) and for evaluation of their antiproliferative effects against HT-29, L-929 and A 549 cell lines by MTT assay at different concentrations (1, 10, 100, 1000 µg/mL). Furthermore, this study aimed to detect antimicrobial activities of the mentioned samples: two Gram-positive (Staphylococcus epidermidis and Staphylococcus aureus), two Gram-negative bacteria (Pseudomonas aeruginosa and Escherichia coli) and a fungi (Candida albicanse), using agar well diffusion method. Results: The IC50 values for antiproliferative activity of the methanolic extract, 20%, 40% and 60% SPE (solid phase extraction) fractions, found to be 345.91 ± 28.77, 442.44 ± 83.22, 220.19 ± 43.13 and 579.90 ± 153.19 μg/mL, respectively; in this case, the maximum inhibition percentage belonged to 40% SPE fraction (220.19 ± 43.13 μg/mL). The total methanolic extract of A. spicigera indicated inhibitory activity against Gram- positive strains, S. epidermidis and S. aureus, with MIC values of 150 mg/mL for both bacteria. Among SPE fractions, 20% was the only active one against Gram possitive species with MIC values of 35 mg/mL for both strains. Conclusions: This study revealed that methanolic extract of A. spicigera and its SPE fractions might be regarded as bioactive natural products, which deserves to be further identification and isolation of cytotoxic and antibacterial phytochemicals from them.

Antiproliferative and apoptotic activities of extracts of Asclepias subulata

Context: Asclepias subulata Decne. (Apocynaceae) is a shrub used in the Mexican traditional medicine for the treatment of cancer.Objective: The objective of this study was to evaluate the antiproliferative activity of methanol extract of aerial parts of A. subulata and its fractions against different cancer cell lines. Additionally, we analyzed the mechanism of action of the active fractions.Materials and methods: Methanol extract fractions were prepared by serial extraction with n-hexane, ethyl acetate, and ethanol. The antiproliferative activity of methanol extract and its fractions was evaluated, against several murine (M12.C3.F6, RAW 264.7, and L929) and human (HeLa, A549, PC-3, LS 180, and ARPE-19) cell lines by the MTT assay, using concentrations of 0.4–400 µg/mL for 48 h. Ethanol and residual fractions were separated using silica gel column. Apoptosis induction of cancer cells was evaluated by Annexin and JC-1 staining using flow cytometry.Results: Methanol extract and its fractions showed antiproliferative activity against all human cancer cell lines tested. Methanol extract had the highest antiproliferative activity on A549 and HeLa cells (IC50 values < 0.4 and 8.7 µg/mL, respectively). Ethanol and residual fractions exerted significant antiproliferative effect on A549 (IC50 < 0.4 µg/mL) and PC3 cells (IC50 1.4 and 5.1 µg/mL). Apoptotic assays showed that CEF7, CEF9, CRF6, and CRF5 fractions induced mitochondrial depolarization in A549 cells, 70, 73, 77, and 80%, respectively. Those fractions triggered the apoptosis mitochondrial pathway.Conclusion: Our data show that A. subulata extracts have potent antiproliferative properties on human cancer cell lines. This plant should be considered an important source of potent anticancer compounds.

Chemical composition and antiproliferative activity of Acalypha californica

Acalypha californica Benth., is a plant in the northwestern region from Mexico, commonly known as “cancer herb” and used in traditional medicine for treating cancer. In the present study we have investigated the antiproliferative activity of methanolic extract of A. californica and its fractions in cancer cell lines and phytochemical analysis and mechanism of apoptosis of the fractions with antiproliferative activity. The antiproliferative activity of methanol extract and its fractions of solvents were evaluated by MTT assay against the M12.Ak.C3.F6, RAW 264.7, HeLa and L929 cell lines. Active fractions were fractionated by molecular exclusion chromatography, HPLC and MPLC. The identification of compounds was performed by NMR and FIA-ESI-IT-MS/MS analysis. Apoptotic mechanism was analyzed by flow cytometry, determining the reduction in the mitochondrial membrane potential (JC-1) and the activity of caspases 3, 8 and 9. Cell viability assays showed that the hexane fraction of the methanol extract of the plant has significant effects against cancer lines RAW 264.7 (IC50=52.08±1.06μg/mL) and HeLa (IC50=46.77±1.09μg/mL), the residual fraction showed a selective effect on cell lines M12.Ak.C3.F6 (IC50=59.90±1.05μg/mL), RAW 264.7 (IC50=58.93±1.26μg/mL) and HeLa (IC50=50.11±1.135μg/mL) compared to the control cell line L929 (IC50=100.00±1.09μg/mL). The chemical characterization of the active fractions allowed the identification of β-sitosterol and stigmasterol in hexane fraction and some phenolic acids, proanthocyanidins and flavonoids in the residual fraction. The methanol extract and hexane fraction reduces mitochondrial membrane potential significantly and activates caspases 3, 8 and 9. Because of the antiproliferative activity observed, our results provide a rational basis for the use of extracts of A. californica in treating various types of cancer in traditional medicine from Mexico. The extracts induce apoptosis via activation of caspases.

Bioactivity-guided isolation of antiproliferative compounds from Centaurea carduiformis DC

In this study, antiproliferative activity of methanol extract of Centaura carduiformis DC. subsp. carduiformis var. carduiformis DC (CC) was examined against Vero, HeLa and C6 cell lines in 1000 and 500μg/mL concentrations using the BrdU ELISA assay. The CC root (CCR) has the most effective antiproliferative activity. The root extract was fractionated using various solvent and determined antiproliferative activities. Two new and two known flavonoids were isolated from CC roots. The isolated known compound of 7-O-β-d-glucopyranosyl-4′-methylapigenin and new compound of 7-O-β-d-6″-O-glucopyranosyl-6″-O-β-d-furanosylpinocembrin were isolated from the ethyl acetate fractions. However, wogonin and new compound of N-(pentyloxy(m-tolyl)methyl)acetamideisowogonin were obtained from chloroform fractions. The chemical structures of pure compounds were elucidated with different chemical and spectroscopic methods (IR, 1H NMR, 13C NMR, HETCOR, COSY, GC-MS, etc.) and their antiproliferative activities were determined against Vero, HeLa and C6 cell lines. The IC50 results showed that the compound 4 has the highest activity against Vero (250μg/mL) and HeLa (735μg/mL) cell lines than isolated other compounds from determined values.

Antiproliferative activity of methanolic extracts from two green algae, Enteromorpha intestinalis and Rizoclonium riparium on HeLa cells

BackgroundNatural compounds can be alternative sources for finding new lead anti-cancer molecules. Marine algae have been a traditional source for bioactive compounds. Enteromorpha intestinalis and Rhizoclonium riparium are two well distributed saline/brackish water algae from Sundarbans. There’s no previous report of these two for their anti-proliferative activities.MethodsCytotoxicity of the algal methanolic extracts (AMEs) on HeLa cells were assayed by 3-(4, 5-dimethylthiazol-2-yl)-2, 5- diphenyltetrazolium bromide (MTT) reduction assay. Morphological examinations were done by Haematoxylin, Hoechst 33258 and Acridine orange staining. DNA fragmentation was checked. Gene expressions of Cysteine aspartate protease (Caspase) 3, Tumor protein (TP) 53, Bcl-2 associated protein X (Bax) were studied by Reverse transcription- polymerase chain reaction (RT-PCR) keeping Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as internal control. Protein expressions were studied for Caspase 3, phospho-p53, Bax, Microtubule associated proteins-1/ light chain B (MAP1/LC3B) by western blot.ResultsThe AMEs were found to be cytotoxic with Inhibitory concentration 50 (IC50) values 309.048 ± 3.083 μg/ml and 506.081 ± 3.714 μg/ml for E. intestinalis and R. riparium extracts respectively. Treated cells became round with blebbings with condensed nuclei. Acidic lysosomal vacuoles formation occurred in treated cells. Expression of apoptotic genes in both mRNA and protein level was lowered. Expression of LC3B-II suggested occurrence of autophagy in treated cells.ConclusionsThese two algae can be potent candidates for isolating new lead anticancer molecules. So they need further characterization at both molecular and structural levels.

Apoptotic Activity of Methanol Extract of Tongcao in HEp-2 Human Cervical Cancer Cells

비록 천연유래물질 중의 하나인 통초가 항산화작용, 항염증작용 및 해열작용과 같은 효능이 있다고 알려져있지만, 종양에서의 암예방효능에 대한 연구는 보고된 바가 없다. 본 연구에서는 사람 자궁경부암세포주인 HEP-2세포에서 통초메탄올추출물의 증식억제 효능 및 관련 분자표적을 확인하고자 하였다. 통초메탄올추출물은 세포증식을 유의성있게 억제하고, 세포사멸을 유도하는 것으로 나타났다. 또한, Bid의 발현에 영향을 주어 truncation을 유도하고, 이로 인해 cytochrome c가 마이토콘드리아에서 세포질로 이동하도록 유도하였지만, Bid 이외의 다른 Bcl-2 family에는 영향을 주지 못했다. 따라서, 이러한 결과를 종합해 볼 때, 통초메탄올추출물은 자궁경부암에서 암예방효능을 가진 잠재성있는 천연추출물이 될 수 있을 것으로 사료된다. Although tongcao has been reported to have anti-oxidative, anti-inflammatory and antipyretic effects, there is no report of the chemopreventive effect of tongcao in cancer cells. In the present study, we investigated the anti-proliferative activity of methanol extract of tongcao (MET) and its molecular target in HEp-2 human cervical cancer cells using MTS assay, western blot analysis, and DAPI staining. MET significantly decreases cell viability and induces apoptotic cell death. It affects Bid protein to be truncated resulting in the release of cytochrome c from mitochondria to cytosol whereas it did not affect other Bcl-2 family members. Thus, we clearly suggest that tongcao can be a potential naturally occurring plants having chemopreventive activity in cervical cancer.

In vitro antioxidant and antiproliferative activities of plants of the ethnopharmacopeia from northwest of Mexico

BackgroundThe aim of this study, is to investigate the in vitro antioxidant activity, the total phenols content, the flavonoids content and the antiproliferative activity of methanolic extracts of the plants: Krameria erecta, Struthanthus palmeri, Phoradendron californicum, Senna covesii and Stegnosperma halimifolium, used by different ethnic groups from northwestern Mexico in the treatment and cure of various diseases.MethodsThe in vitro antioxidant activity was measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and Ferric Reducing/Antioxidant Power assay (FRAP), the total phenols content was measured by Folin–Ciocalteau assay, the flavonoids content by the AlCl3 colorimetric method and the antiproliferative activity (line cells HeLa, RAW 264.7, M12Ak.C3.F6 and L929) using MTT method.ResultsThe K. erecta extract showed the higher radical scavenging activity (67.88%), antioxidant activity by FRAP (1.41 mg Trolox Eq), the highest total phenols content (598.51 mg Galic Acid Eq/g extract), the highest flavonoids content (3.80 mg Quercetin Eq/g extract) and the greatest antiproliferative activity in a dose dependent manner against most Cell line evaluated. A positive correlation was found between the antioxidant activity and the flavonoids content.ConclusionsThis study is the first report on the antioxidant and antiproliferative activities of the five species evaluated. The results demostrate that there is a positive correlation between antioxidant activity and the flavonoids content, indicating that these type of polyphenols could be the major contributors to the observed antioxidant activity in the evaluated plant extracts. Of the extracts evaluated, that of Krameria erecta showed the greatest antioxidant and antiproliferative activities, a discovery that makes this species a promising candidate for future research.

Antiproliferative activity of pentadeca-(8E, 13Z) dien-11-yn-2-one and (E)-1,8-pentadecadiene from Echinacea pallida (Nutt.) Nutt. roots

Several species of Echinacea, a perennial plant which belongs to the Asteraceae family, possess medicinal properties and are currently used in phytotherapy. In the present study, antiproliferative activity of methanol extract and isolated structures of pentadeca-(8E, 13Z)-dien-11-yn-2-one 1 and (E)-1,8-pentadecadiene 2 from Echinacea pallida roots on C6 cells (Rat Brain tumor cells) and HeLa cells (human uterus carcinoma) was investigated in vitro. Antiproliferative effect of the extract, isolated compounds, and cisplatin were tested at 5, 10, 20, 30, 40, 50, 75, and 100 μg ml−1 using BrdU Cell Proliferation ELISA. The methanol extract and Compound 1 significantly inhibited proliferation of HeLa and C6 cancer cell lines.

Teucrium Plant Species as Natural Sources of Novel Anticancer Compounds: Antiproliferative, Proapoptotic and Antioxidant Properties

This study deals with total phenolic content, antiproliferative and proapoptotic activity of methanolic extracts from different Teucrium species and the effect on the prooxidant/antioxidant status in HCT-116 cells. The total phenolic content of the extracts was measured spectrophotometricaly and the obtained results ranged from 56.62 mg/g to 172.50 mg GA/g. The antiproliferative activity of methanolic extracts from different Teucrium species was determined using MTT cell viability assay, where IC50 value was used as a parameter for cytotoxicity. The type of cell death was explored by fluorescence microscopy using the acridin orange/ethidium bromide method. MTT assay showed that all extracts significantly reduced cell viability in a dose-dependent manner, with very low IC50 values. The highest content of phenolic compounds and the best cytotoxic activity on HCT-116 cells after 24 h of exposure was in T. chamaedrys extract, with IC50 values of 5.48 × 10−9 μg/mL. After 72 h, methanolic extract of T. arduini appeared to have the best cytotoxic activity on HCT-116, with IC50 values of 0.37 μg/mL. Treatments caused typical apoptotic morphological changes in HCT-116 cells and showed a high percentage of apoptotic cells. The results of the presented research indicate that some Teucrium extracts are a very rich source of phenols, which may directly contribute to high antiproliferative and proapoptotic activity.