Apoptogenic Properties Research Articles

Cobalt oxide nanoparticles functionalized with glucose and conjugated to Ellagic acid (Co3O4@Glu-Ellagic acid NPs) induce apoptotic genes in HepG2, liver cancer cell line

Surface modification of metal nanoparticles can be a solution to obtain more efficient and biocompatible drugs. In this study, the anticancer mechanism of cobalt oxide nanoparticles (NPs) functionalized with glucose and conjugated with Ellagic acid (Co3O4@Glu-Ellagic acid NPs) in a liver cancer cell line was investigated. Physicochemical characteristics of NPs were investigated by Fourier-transform infrared spectroscopy (FT-IR), Transmission electron microscopy (TEM), (Scanning Electron Microscopy) SEM, X-Ray diffraction analysis (XRD), Energy Dispersive Spectroscopy mapping (EDS-mapping), Dynamic light scattering (DLS), and zeta potential. Cytotoxicity of Co3O4@Glu-Ellagic acid NPs in normal (THLE2) and cancer (HepG2) hepatocyte cell lines was characterized by MTT and flow cytometry assays. Relative expression of the CASP8, CASP9, and p53 genes was assessed by Real-Time PCR. The synthesized nanoparticles were spherical, without considerable agglomeration, and their size was in the range of 20 to 60 nm. Surface charge and DLS size were − 30.1 mV and 280 nm, respectively. Treatment of the studied cells with the nanoparticles displayed their antiproliferative potential; however, liver cancer cells were considerably more sensitive to the nanoparticles than normal hepatocyte cells. The 50 % inhibitory concentration of Co3O4@Glu-Ellagic acid NPs for normal and cancer hepatocyte cells were 201 and 97 μg/mL, respectively. Treating HepG2 cells with Co3O4@Glu-Ellagic acid NPs led to a significant increase in the percentage of apoptotic cells (from 0.66 % to 50.8 %) and expression of the CASP8, CASP9, and p53 genes by 2.88, 2.11, and 2.0 folds that suggests apoptogenic property of the nanoparticles through activation of the intrinsic and extrinsic apoptosis pathways. Measuring Reactive Oxygen Species (ROS) level showed that in treated cancer cells with Co3O4@Glu-Ellagic acid nanoparticles the ROS level increased. Moreover, Hoechst staining indicated the anti-tumor effect of Co3O4@Glu-Ellagic acid NP by appearance of apoptotic bodies in HepG2 cells. This work revealed that Co3O4@Glu-Ellagic acid NPs are potent apoptogenic agent against liver cancer cells.

Therapeutic Potential of Chlorophyll Functionalized Carbon Quantum Dots against Cervical Cancer

AbstractCervical cancer is an emerging gynecological disorder that affects the morbidity and mortality of the females worldwide. In 2018, according to the World Health Organization, 570,000 women were diagnosed with cervical cancer and 311,000 women died from the condition. Various anticancer agents are in the run to combat cervical cancer but overall results are still demoralizing. Various reports suggest that Quantum dots are being used as promising agents against several types of cancer. In the current study, we synthesized high‐quantum yield chlorophyll functionalized carbon quantum dots (Chl‐CQDs) from banana leaves with a simple and environment friendly hydrothermal process. The free radical activity of Chl‐CQDs was measured using diphenyl picrylhydrazyl (DPPH) and azino‐bis (ethylbenzothiazoline sulphonic acid) diammonium salt (ABTS⋅+), as well as their cytotoxic, bioimaging, and apoptogenic properties were also evaluated on normal cell line HEK‐293 and human cancer cell lines SiHa. Our results of dimethylthiazol diphenyltetrazolium bromide tetrazolium reduction (MTT) assay showed that Chl‐CQDs have profound effect on SiHa cells with an IC50 of approximately 100 μg/ml after 24 hours and no effect HEK‐293 cells. Bioimaging and apoptogenic assays confirmed that Chl‐CQDs exhibited strong therapeutic potential against SiHa cells and no cytotoxic potential against HEK‐293 cells. The cumulative findings suggested that Chl‐CQDs exhibited non‐cytotoxic nature against HEK‐293 cells whereas cytotoxic nature against SiHa cells.

Chemical Characterization, In-silico Evaluation, and Molecular Docking Analysis of Antiproliferative Compounds Isolated from the Bark of Anthocephalus cadamba Miq.

The present study aimed to isolate and characterize chemical compounds from Anthocephalus cadamba Miq. bark and evaluate their anticancer activity by in silico, molecular docking, and in vitro studies. Anthocephalus cadamba is a traditionally used Indian medicinal plant. The anticancer and phytochemical properties of this plant remain unexplored except for a few studies. The objective of the study was to evaluate the antiproliferative activity of extract and fractions against breast cancer and prostate cancer cell lines and isolate and characterize active compounds from bio-active guided fractions. Moreover, the anticancer activity of isolated compounds against breast and prostate cancer cell lines was also evaluated, in addition to in silico and molecular docking interactions of isolated compounds with VEGFR2 and PDGFRα target proteins. The compounds were isolated and purified with the help of repeated column chromatography, and spectral techniques, such as 1D, 2D NMR, and GC-MS/MS, were used to identify and elucidate the structure of the compounds. Moreover, prediction of activity spectra for substances, physiochemical properties, bioactivity radar prediction, bioactivity score, natural-product likeness, ADME, and toxicity parameters of isolated compounds (AC-1 to AC-4) was performed through various in-silico databases and servers. To evaluate the docking interaction profile and binding energies of compounds, three docking tools were utilized, such as AutoDock, AutoDock Vina, and iGEMDOCK, against two targets VEGFR2 and PDGFRα. MD simulation was performed through ligand and receptor molecular dynamic server (LARMD). It was found that the A. cadamba bark chloroform fraction demonstrated a significant inhibitory effect against MDA-MB-231, MCF-7, and PC-3 cells in a dose-time-dependent manner. The bioassay-guided isolation afforded four molecules AC-1 to AC-4 from chloroform fraction. Moreover, the GC-MS/MS profiling identified fourteen new molecules which were not reported earlier from A. cadamba. The in-silico study showed that the isolated compounds (AC-1 to AC-4) followed Lipinski's rule and had good oral bioavailability. While compound AC-4 had positive bioactivity scores except for kinase inhibitor activity. The ADMET profiling revealed that AC-4 was non-toxic and easily absorbed in the human intestine, and transportable in the blood-brain barrier compared to AC-1, AC-2, AC-3, and standard drug doxorubicin. Molecular docking and MD simulation assessment also signified AC-4 anticancer activity with dual inhibitory action against the target proteins VEGFR2 and PDGFRα amongst the studied compounds. The in vitro cell viability assay of isolated compounds demonstrated that AC-1 showed IC50 (μg/mL) value of 34.96 ±3.91, 47.76±3.80 69.1±4.96, AC-2; 68.26±4.22, 54.03±5.14, >100, AC-3; 35.34±4.14, 51.5±51.5, 70.8±5.25 and AC-4; 44.2±3.57, 24.2±2.67, 51.2±2.54 for MDA-MB-231, MCF-7, and PC-3 cancer cell lines, respectively and compared with standard drug doxorubicin. Moreover, fluorescence microscopy confirmed the apoptogenic property of compounds. We also found that AC-4 exhibited significant intracellular ROS production in breast cancer cells, thereby inducing apoptosis and eventually cell death. In conclusion, A. cadamba afforded four pure molecules AC-1 to AC-4 with the identification of fourteen new compounds. The entire in-silico studies concluded that the AC-4 compound had better oral bioavailability, bioactivity score, and ADMET profile among studied molecules. Molecular docking analysis and MD simulation also supported AC-4 dual inhibitory action against both VEGFR2 and PDGFRα receptors. Moreover, the isolated molecules AC-1, AC-2, AC-3, and AC-4 were found to be active against MDA-MB-231, MCF-7, and PC-3 cancer cells. The molecule AC-4 was found to induce ROS-mediated apoptosis in breast cancer cells. It was found that the anticancer inhibitory potentiality of AC-4 is directed to its molecular stereochemistry which specifically binds to the target proteins of breast cancer cells with no toxicological effect. Therefore, AC-4 is suggested to be an effective aspirant for novel drug design and discovery.

Evaluating the antimicrobial and apoptogenic properties of bacteriocin (nisin) produced by Lactococcus lactis

In this study, we extracted nisin from Lactococcus lactis, purified it, and examined it for biological properties such as antibacterial and anticancer activities. The purified nisin was characterised using powder X-ray diffraction (XRD), fourier transform infrared (FT-IR), and UV-Vis spectroscopy. When purified nisin was tested for antibacterial and antifungal activity against various bacterial and fungal cultures, it was found to have superior antibacterial and antifungal abilities compared to earlier literature. Superior antibacterial activity was confirmed via intracellular ROS generation. Further, MTT studies on a variety of cancer and normal cell lines, including HepG2, HeLa, A549, HDFa, and THLE-2, show that purified nisin has high anticancer activity while causing negligible damage to normal cell lines. The findings pave the path for new anti-cancer drugs based on the nisin-based therapy approach in the future.

Anti-breast cancer potential of MnO2 nanoparticles using Terminalia chebula fruit extract against MCF-7 cell line through in vitro cell cycle and apoptotic studies

Since decades, cancer has become reason of major concern as it is the leading cause of mortality worldwide. Many therapeutic strategies have come up in the scientific world, but its pitiful to know that synthetic chemotherapeutic agents either cause adverse effects or cancer cells develop resistance to these agents. Plant derived chemotherapeutic agent present wide range of therapeutics, and most are yet to be discovered. In the present study, we have analysed the anti-proliferative, and apoptogenic properties of manganese oxide nanoparticles MnO2 NPs derived from Terminalia chebula (TC) fruit on breast adenocarcinoma, MCF7 cell lines. The formation of MnO2NPs was confirmed through scanning electron microscopy (SEM), transmission electron microscopy (TEM) and high-resolution transmission electron microscopy (HR-TEM). The morphology of the material was studied using SEM analysis, which suggested a agglomeration having spherical shape with an average diameter of 47.6 nm. Further, the TEM and HR-TEM images confirmed the rod shape of the as-prepared MnO2 NPs with an interplanar distance of 0.271 nm. The anticancer efficacy of MnO2NPs was evaluated against MCF-7 breast cancer cell line, which showed up to 86% inhibition of the cells at 320 μg/mL concentration.The MnO2 NPs was found to be cytotoxic, and was able to induce apoptosis in MCF7 cell lines.

Modulation of Bax and Bcl-2 genes by secondary metabolites produced by Penicillium rubens JGIPR9 causes the apoptosis of cancer cell lines

ABSTRACT Search for an efficient anti-cancer compound of natural origin with well-defined mechanisms of action is an important scientific pursuit today, due to cancer being the second leading cause for the death of affected people. The members of the genus Penicillium are one of the important sources of bioactive compounds. In the present study, Penicillium rubens, isolated from a garden soil in Madurai district of Tamil Nadu, was found to produce a highly promising anti-cancer metabolite. The percentage viabilities of HepG2, HeLa and MCF-7 cancer cells treated with the bioactive fraction (P5) isolated from P. rubens, ranged between 40-50% after 96 h. Apoptosis induction was found to be the major reason for the observed reduction in cancer cell proliferation and cell count which was confirmed by caspase activity, DNA fragmentation, clonogenic assay, cell cycle analysis and LDH assays. The upregulation of proapoptotic Bax, coupled with the downregulation of anti-apoptotic Bcl-2 expressions were confirmed by RT-qPCR and flow cytometry methods. The current study also indicated an upregulation of p53 which further strengthened the apoptogenic property of P5 fraction. Non-toxicity of P5 was demonstrated on normal peripheral lymphocytes. The analysis of P5 fraction through GC-MS indicated the presence of indole-2, 3-(4,4-dimethyl-3-thiosemicarbazone) as one of the major compounds.

Cytotoxicity and apoptogenic properties of the standardized extract of Portulaca oleracea on glioblastoma multiforme cancer cell line (U-87): a mechanistic study.

The traditional uses of Portulaca oleracea L. (PO) with anti-inflammatory and anti-cancer activity as well as antioxidants properties were expressed previously. Glioma is considered the most common primary brain tumor and its malignant form is the most lethal adult brain tumor, that glioblastoma covers about 50 % of glioma tumors. The present study was aimed to evaluate the cytotoxicity and apoptogenic effects of the hydro-ethanolic extract of PO on human glioblastoma cancer cell line (U-87) and the role of NF-κB. Cytotoxicity of the extract in the presence or absence of Vitamin C was evaluated using MTT assay, and the following hypotonic PI and SubG1 peak were performed. Moreover, the reactive oxygen species (ROS), the level of NF-κB protein and nitric oxide (NO) production were investigated. The extract had cytotoxicity and apoptogenic effects on U-87 cells in both the concentration and time-dependent manners. The mechanism of cytotoxicity and apoptosis induction of the extract at the first hours of incubation and low concentrations were dependent on ROS. However, the toxicity was replaced with NO pathway with time-lapse and higher concentrations. Results also indicated that the extract acts as an NF-κB inhibitor with concentration and time-dependent manners. The present study may suggest the anti-NF-κB activity of PO along with two upstream ROS and NO mechanisms. Furthermore, the extract as ethnobotanical may be used as adjunctive anti-cancer therapy against glioblastoma multiforme.

Measles virus phosphoprotein inhibits apoptosis and enhances clonogenic and migratory properties in HeLa cells

Measles virus is the causative agent of measles, a major cause of child mortality in developing countries. Two major proteins, coded by the viral genome, are nucleocapsid protein (N) and phosphoprotein (P). The N protein protects the viral genomic RNA and forms ribonucleoprotein complex (RNP) together with P protein. MeV-P protein recruits the large protein (L), i.e. viral RNA-depended RNA polymerase (RdRp), to ensure viral replication in host cell. Apoptogenic properties of N protein of Edmonston vaccine strain have been established in our lab previously. We investigated the role of MeV-P protein of Edmonston vaccine strain as modulator of apoptosis in cervical cancer cell line (HeLa) and found that MeV-P protein is anti-apoptotic and enhances cell proliferation. Measles virus is considered to be innately oncotropic virus. However, the anti-apoptotic property of MeV-P protein raises important concerns while adopting this virus as an anti-cancer therapeutic tool.

Long-term tumor-free survival in a metastatic pancreatic carcinoma patient with FOLFIRINOX/Mitomycin, high-dose, fever inducing Viscum album extracts and subsequent R0 resection

Rationale:Metastatic pancreatic cancer has a dismal prognosis. Many patients seek integrative care as an add-on to their conventional cancer treatment. Viscum album extracts (VAE)—widely used as an adjunct to cancer treatment—have cytotoxic, apoptogenic, and immune stimulatory properties. A statistically significant survival benefit has been demonstrated for VAE in advanced pancreatic cancer.Patient concerns and diagnosis:A 28-year old patient presented with painless jaundice and was subsequently diagnosed as pancreatic adenocarcinoma with liver metastases.Interventions:He was treated with FOLFIRINOX/Mitomycin, hyperthermia and fever-inducing VAE.Outcomes:Subsequently, the liver metastases regressed. Surgical intervention involved successful R0-resection of the primary tumor, as well as an atypical liver resection. A relapse was again treated with FOLFIRINOX/Mitomycin and hyperthermia. As of publication of this report, 49 months after initial diagnosis, the patient exhibits good condition, and is unrestricted in quality of life (till publication).Lessons:This case demonstrates the favorable outcome of a patient with metastatic pancreatic cancer following treatment with chemotherapy, integrative medicine, and surgical excision. As other positive outcomes in pancreatic cancer patients are related to inflammatory events, we presume the immunologic effects of VAE to have contributed to the favorable outcome here. Based on this case, and the other positive results of VAE use in pancreatic cancer, further investigations seem highly worthwhile.

Green Synthesis of Carbon Dots Derived from Walnut Oil and an Investigation of Their Cytotoxic and Apoptogenic Activities toward Cancer Cells.

Purpose: This paper introduces a green and simple hydrothermal synthesis to prepare carbon quantum dots (CQDs) from walnut oil with a high quantum yield. In addition, cytotoxic and apoptogenic properties of the CQDs were analyzed on human cancer cell lines.Methods: The optical properties and morphological characteristic were investigated by the TEM, XRD, FT-IR, UV-vis and photoluminescence (PL).The cytotoxic potential of walnut CQDs was evaluated on PC3, MCF-7 and HT-29 human carcinoma cell lines using the MTT methods. The mechanism of action was studied by investigating the mode of cell death using the activation of caspase-3 and 9 as well as mitochondrial membrane potential (MMP). Cellular uptake of the CQDs was detected by fluorescence microscope. CQDs had an average size of 12 nm and a significant emission at 420 nm at an excitation wavelength of 350 nm was recorded.Results: The prepared CQDs possessed a good fluorescent quantum yield of 14.5% with quinine sulfate (quantum yield 54%) as a reference and excellent photo as well as pH stabilities. The walnut CQDs were proved to be an extremely potent cytotoxic agent, especially against MCF-7 and PC-3 cell lines. Induction of apoptosis by CQDs was accompanied by an increase in the activation of caspase-3. Caspase-9 activity did not increase after exposure to the CQDs. Additionally; the MMP did not show any significant loss.Conclusion: The results of our study can corroborate the cytotoxic and apoptotic effect of walnut CQDs in the PC3 and MCF-7 cancer cell lines.

Peculiarities of modification by astaxanthin of radiation-induced damages in the genome of human blood lymphocytes exposed in vitro on different stages of the mitotic cycle

The features of astaxanthin impact (20 μg/mL) in the culture of human blood lymphocytes exposed to γ-radiation (1.0 Gy) on the G0, S, and G2 phases of the cell cycle were studied using Comet assay. Decrease in the level of DNA damages (Tail Moment index) under astaxanthin influence on lymphocytes irradiated in all stages of cell division was established, while, as a result of previous cytogenetic investigations, lack of the modifying action of astaxanthin after irradiation of cells in the G2 stage and radioprotective effect in the G0 stage of the mitotic cycle had been revealed. In G0 phase, the activation of the processes of apoptosis by astaxanthin in irradiated cells with high levels of genomic damages was found. The obtained data demonstrate that astaxanthin has a powerful radioprotective potential, mainly due to its apoptogenic properties.

Anticancer mechanisms of Strobilanthes crispa Blume hexane extract on liver and breast cancer cell lines.

Cancer is a major public health concern not only in developed countries, but also in developing countries. It is one of the leading causes of mortality worldwide. However, current treatments may cause severe side effects and harm. Therefore, recent research has been focused on identifying alternative therapeutic agents extracted from plant-based sources in order to develop novel treatment options for cancer. Strobilanthes crispa Blume is a plant native to countries including Madagascar and Indonesia. It has been used as an anti-diabetic, diuretic and laxative in traditional folk medicine. Furthermore, S. crispa has potential in treating cancer, as evidenced in previous studies. In the present study, the cytotoxic and apoptotic activities of S. crispa crude extracts were investigated in liver and breast cancer cell lines. Hexane, ethyl acetate, chloroform, methanol and water extracts prepared from the leaves, and stems of S. crispa were evaluated for their cytotoxicity on HepG-2 and MDA-MB-231 cells using an MTT assay. The anti-proliferative properties of stem hexane (SH) extract on both cell lines were analysed using cell doubling time determination and cell cycle analysis, while the apoptogenic properties was determined through the detection of caspase-8. Among the extracts tested, SH extract exhibited the lowest half maximal inhibitory concentrations in both the cell lines. The SH extract induced morphological changes in HepG-2 and MDA-MB-231 cells, and significantly delayed cell population doubling time. Furthermore, it altered cell cycle profile and significantly increased caspase-8 activity in HepG-2 cells, but not in MDA-MB-231 cells. In conclusion, the SH extract of S. crispa possesses potent anticancer properties and may be a suitable chemotherapeutic target.

Standardized Polyalthia longifolia leaf extract (PLME) inhibits cell proliferation and promotes apoptosis: The anti-cancer study with various microscopy methods.

Over the years a number of microscopy methods have been developed to assess the changes in cells. Some non-invasive techniques such as holographic digital microscopy (HDM), which although does not destroy the cells, but helps to monitor the events that leads to initiation of apoptotic cell death. In this study, the apoptogenic property and the cytotoxic effect of P. longifolia leaf methanolic extract (PLME) against the human cervical carcinoma cells (HeLa) was studied using light microscope (LM), holographic digital microscopy (HDM), scanning electron microscope (SEM) and transmission electron microscope (TEM). The average IC50 value of PLME against HeLa cells obtained by MTT and CyQuant assay was 22.00μg/mL at 24h. However, noncancerous Vero cells tested with PLME exhibited no cytotoxicity with the IC50 value of 51.07μg/mL at 24h by using MTT assay. Cytological observations showed nuclear condensation, cell shrinkage, multinucleation, abnormalities of mitochondrial cristae, membrane blebbing, disappearance of microvilli and filopodia, narrowing of lamellipodia, holes, formation of numerous smaller vacuoles, cytoplasmic extrusions and formation of apoptotic bodies as confirmed collectively by HDM, LM, SEM and TEM. In conclusion, PLME was able to produce distinctive morphological features of HeLa cell death that corresponds to apoptosis.

225 - Expansion of Bone Marrow NK Lymphocytes is Associated with Higher Probability of Response to Azacitidine Treatment and Survival Benefit

Over the years a number of microscopy methods have been developed to assess the changes in cells. Some non-invasive techniques such as holographic digital microscopy (HDM), which although does not destroy the cells, but helps to monitor the events that leads to initiation of apoptotic cell death. In this study, the apoptogenic property and the cytotoxic effect of P. longifolia leaf methanolic extract (PLME) against the human cervical carcinoma cells (HeLa) was studied using light microscope (LM), holographic digital microscopy (HDM), scanning electron microscope (SEM) and transmission electron microscope (TEM). The average IC50 value of PLME against HeLa cells obtained by MTT and CyQuant assay was 22.00 μg/mL at 24 h. However, noncancerous Vero cells tested with PLME exhibited no cytotoxicity with the IC50 value of 51.07 μg/mL at 24 h by using MTT assay. Cytological observations showed nuclear condensation, cell shrinkage, multinucleation, abnormalities of mitochondrial cristae, membrane blebbing, disappearance of microvilli and filopodia, narrowing of lamellipodia, holes, formation of numerous smaller vacuoles, cytoplasmic extrusions and formation of apoptotic bodies as confirmed collectively by HDM, LM, SEM and TEM. In conclusion, PLME was able to produce distinctive morphological features of HeLa cell death that corresponds to apoptosis.

Bee venom induces apoptosis and suppresses matrix metaloprotease-2 expression in human glioblastoma cells

Glioblastoma is the most common malignant brain tumor representing with poor prognosis, therapy resistance and high metastasis rate. Increased expression and activity of matrix metalloproteinase-2, a member of matrix metalloproteinase family proteins, has been reported in many cancers including glioblastoma. Inhibition of matrix metalloproteinase-2 expression has resulted in reduced aggression of glioblastoma tumors in several reports. In the present study, we evaluated effect of bee venom on expression and activity of matrix metalloproteinase-2 as well as potential toxicity and apoptogenic properties of bee venom on glioblastoma cells. Human A172 glioblastoma cells were treated with increasing concentrations of bee venom. Then, cell viability, apoptosis, matrix metalloproteinase-2 expression, and matrix metalloproteinase-2 activity were measured using MMT assay, propidium iodide staining, real time-PCR, and zymography, respectively. The IC50 value of bee venom was 28.5μg/ml in which it leads to decrease of cell viability and induction of apoptosis. Incubation with bee venom also decreased the expression of matrix metalloproteinase-2 in this cell line (p<0.05). In zymography, there was a reverse correlation between bee venom concentration and total matrix metalloproteinase-2 activity. Induction of apoptosis as well as inhibition of matrix metalloproteinase-2 activity and expression can be suggested as molecular mechanisms involved in cytotoxic and antimetastatic effects of bee venom against glioblastoma cells.

Cytotoxic and apoptogenic effects of Bryonia aspera root extract against Hela and HN-5 cancer cell lines.

Bryonia aspera (Stev. ex Ledeb) is a plant that grows in northeast of Iran. In the present study, cytotoxic and apoptogenic properties of B. aspera root extract was determined against HN-5(head and neck squamous cell carcinoma) and Hela (cervix adenocarcinoma) cell lines. HN-5 and Hela cell lines were cultured in DMEM medium and incubated with different concentrations of B. aspera root extract. Cell viability was quantitated by MTT assay and the optical absorbance was measured at 570 nm (620 nm as the reference) by an ELISA reader, in each experiment. Apoptotic cells were assessed using PI staining of DNA fragmentation by flow cytometry (sub-G1 peak). The B. aspera inhibited 50% growth (IC50) of Hela and HN-5 cell lines at 100±28 μg/ml and 12.5±4 μg/ml, respectively after 48 hr of incubation. Cell viability assay showed that inhibitory effects of B. aspera were time and dose-dependent in both cell lines, which were consistent with morphological changes, observed under light microscope. Apoptosis was investigated by flow cytometry in which percentage of apoptotic cells increased in a dose and time-dependent manner. Based on our data, B. aspera has cytotoxic effects in which apoptosis played an important role. Further evaluations are needed to assess the possible anti-tumor properties of this plant.

Effects of Biebersteinia multifida hydro-ethanol extract on proliferation and apoptosis of human prostate cancer and human embryonic kidney cells.

Biebersteinia (Geraniaceae) has a history of use in traditional medicine in some countries including Iran. In the present study, cytotoxic and apoptogenic properties of hydro-ethanol extract of B. multifidi was investigated on human prostate cancer cell lines (PC3 and DU 145) and human embryonic kidney 293(HEK293) cells. Cells were cultured in RPMI-1640 medium supplemented with 10% FBS at 37ºC in a humidified atmosphere of 95% air and 5% CO2. The root of the plant was macerated with EtOH 70%. Cytotoxic activity of ethanol extract of B. multifida was assessed using alamarBlue® assay after 48 hr of treatment. Apoptotic cells were stained with propidium iodide (PI) and detected by flow cytometry (sub-G1 peak). B.multifidi had cytotoxic effect on malignant cells and normal HEK293 cells in a dose-dependent manner and significantly decreased the cell viability (IC50 values were between 199.2 and 302.9 µg/ml). B. multifida increased the sub-G1 peak in flow cytometry histogram of treated PC3 cells compared to control showing the induction of apoptosis and DNA fragmentation. Due to cytotoxic and apoptotic activity of B. multifida, the plant is suggested for further phytochemical analysis and mechanistic evaluation.

Medical application of scorpion venom to breast cancer: A mini-review

Breast cancer is the leading cause of mortality in women worldwide. Today, 1 in 8 women born in the United States will have an invasive cancer in their lifetime. Despite significant attempts, the prognosis of metastatic breast cancer still remains poor. This has compelled scientists to look elsewhere for better therapeutic outcomes. Recent advances in venomic studies have demonstrated some promise in cancer-related ailments. Scorpion venom, a complex cocktail of biogenic amines, proteins, peptides, mucoproteins, organic salts and neurotoxins has shown a potential therapeutic application due to its cytotoxic, apoptogenic, immunosuppressive and antiproliferative properties. This communication reviews the effects of scorpion venom components on breast cancer and their mechanisms.

Cytotoxicity of Wortmannin Triggers Programmed Cell Death in MCF-7 Cells; Biochemical and Morphological Analysis

Objective: The primary objective was to study whether the cytotoxicity of Wortmannin (Wtmn) triggers apoptotic programmed cell death on the widely-used model breast cancer MCF-7 cell to understand this small molecule’s biological signaling mechanisms. Methods: In this present study, MCF-7 cells anti-proliferative activities determined by using MTT assays to quantify the cytotoxicity of the Wtmn. The direct cellular and molecular effect of Wtmn was investigated selectively using flow cytometric cell cycle, mitochondrial membrane potential (MMP) assays, fluorescent and phase contrast microscopy. Results: Our biochemical and morphological experimental analysis showed that Wtmn inhibits the proliferation of MCF-7 breast cancer cells and facilitates their entry into the apoptosis mode of programmed cell death. Conclusion: Our experimental results directed towards elucidating the cytotoxicity of Wtmn and it’s biological mechanisms indicates anti-proliferative and apoptogenic properties. This strategy can serve as a screening model for anti-cancer treatment in-vitro.

Cleistopholine isolated from Enicosanthellum pulchrum exhibits apoptogenic properties in human ovarian cancer cells

BackgroundCleistopholine is a natural alkaloid present in plants with numerous biological activities. However, cleistopholine has yet to be isolated using modern techniques and the mechanism by which this alkaloid induces apoptosis in cancer cells remains to be elucidated. Hypothesis/purposeThis study aims to isolate cleistopholine from the roots of Enicosanthellum pulchrum by using preparative-HPLC technique and explore the mechanism by which this alkaloid induces apoptosis in human ovarian cancer (CAOV-3) cells in vitro from 24 to 72h. This compound may be developed as an anticancer agent that induces apoptosis in ovarian cancer cells. Study design/methodsCytotoxicity was assessed via the cell viability assay and changes in cell morphology were observed via the acridine orange/propidium iodide (AO/PI) assay. The involvement of apoptotic pathways was evaluated through caspase analysis and multiple cytotoxicity assays. Meanwhile, early and late apoptotic events via the Annexin V-FITC and DNA laddering assays, respectively. The mechanism of apoptosis was explored at the molecular level by evaluating the expression of specific genes and proteins. In addition, the proliferation of CAOV-3-cells treated with cleistopholine was analysed using the cell cycle arrest assay. ResultsThe IC50 of cleistopholine (61.4µM) was comparable with that of the positive control cisplatin (62.8µM) at 24h of treatment. Apoptosis was evidenced by cell membrane blebbing, chromatin compression and formation of apoptotic bodies. The initial phase of apoptosis was detected at 24h by the increase in Annexin V-FITC binding to cell membranes. A DNA ladder was formed at 48h, indicating DNA fragmentation in the final phase of apoptosis. The mitochondria participated in the process by stimulating the intrinsic pathway via caspase 9 with a reduction in mitochondrial membrane potential (MMP) and an increase in cytochrome c release. Cell death was further validated through the mRNA and protein overexpression of Bax, caspase 3 and caspase 9 in the treated cells compared with the untreated cells. In contrast, Bcl-2, Hsp70 and survivin decreased in expression upon cleistopholine treatment. Cell cycle was arrested at the G0/G1 phase and cell population percentage significantly increased to 43.5%, 45.4% and 54.3% in time-dependent manner in the cleistopholine-treated CAOV-3 cells compared with the untreated cells at 24, 48 and 72h respectively. ConclusionThe current study indicated that cleistopholine can be a potential candidate as a new drug to treat ovarian cancer disease.