Cell Line 4T1 Research Articles

HSA-Binding Prodrugs-Based Nanoparticles Endowed with Chemo and Photo-Toxicity against Breast Cancer.

Simple SummaryIn this study, we developed novel bioresponsive HSA-binding nanoparticles co-delivering paclitaxel (PTX) prodrugs and the photosensitizer pheophorbide a (Pba) for the combined photo- and chemo-treatment of breast cancer. Extensive structural characterization allowed us to evaluate the size, stability and morphology of nanoparticles, which exhibited sustained and controlled drug release under the distinctive redox conditions of the tumor environment. HSA-binding PTX/Pba nanoparticles showed higher Pba uptake in human breast cancer cells and a synergistic antitumor effect upon light irradiation. Preliminary in vivo experiments using low drug doses showed the potential of our bioresponsive nanoparticles to reduce the primary tumor mass while diminishing the number of lung metastases, thus suggesting the effectiveness of this novel approach.Exploiting the tumor environment features (EPR effect, elevated glutathione, reactive oxygen species levels) might allow attaining a selective and responsive carrier capable of improving the therapeutic outcome. To this purpose, the in situ covalent binding of drugs and nanoparticles to circulating human serum albumin (HSA) might represent a pioneering approach to achieve an effective strategy. This study describes the synthesis, in vitro and in vivo evaluation of bioresponsive HSA-binding nanoparticles (MAL-PTX2S@Pba), co-delivering two different paclitaxel (PTX) prodrugs and the photosensitizer pheophorbide a (Pba), for the combined photo- and chemo-treatment of breast cancer. Stable and reproducible MAL-PTX2S@Pba nanoparticles with an average diameter of 82 nm and a PTX/Pba molar ratio of 2.5 were obtained by nanoprecipitation. The in vitro 2D combination experiments revealed that MAL-PTX2S@Pba treatment induces a strong inhibition of cell viability of MDA-MB-231, MCF7 and 4T1 cell lines, whereas 3D experiments displayed different trends: while MAL-PTX2S@Pba effectiveness was confirmed against MDA-MB-231 spheroids, the 4T1 model exhibited marked resistance. Lastly, despite using a low PTX-PDT regimen (e.g., 8.16 mg/Kg PTX and 2.34 mg/Kg Pba), our formulation showed to foster primary tumor reduction and curb lung metastases growth in 4T1 tumor-bearing mice, thus setting the basis for further preclinical validations.

Amphiphilic Protoporphyrin IX Derivatives as New Photosensitizing Agents for the Improvement of Photodynamic Therapy.

Photodynamic therapy (PDT) is a non-invasive therapeutic modality based on the interaction between a photosensitive molecule called photosensitizer (PS) and visible light irradiation in the presence of oxygen molecule. Protoporphyrin IX (PpIX), an efficient and widely used PS, is hampered in clinical PDT by its poor water-solubility and tendency to self-aggregate. These features are strongly related to the PS hydrophilic–lipophilic balance. In order to improve the chemical properties of PpIX, a series of amphiphilic PpIX derivatives endowed with PEG550 headgroups and hydrogenated or fluorinated tails was synthetized. Hydrophilic–lipophilic balance (HLB) and log p-values were computed for all of the prepared compounds. Their photochemical properties (spectroscopic characterization, photobleaching, and singlet oxygen quantum yield) were also evaluated followed by the in vitro studies of their cellular uptake, subcellular localization, and photocytotoxicity on three tumor cell lines (4T1, scc-U8, and WiDr cell lines). The results confirm the therapeutic potency of these new PpIX derivatives. Indeed, while all of the derivatives were perfectly water soluble, some of them exhibited an improved photodynamic effect compared to the parent PpIX.

In Vivo Investigation of Supportive Immunotherapeutic Combination of Bifidobacterium infantis 35624 and Doxorubicin in Murine Breast Cancer.

The aim of the study is to investigate the anti-tumor effect of Bifidobacterium infantis 35624 in a xenograft model in BALB/c miceinjected with 4T1 cells as a support for chemotherapeutic treatments of doxorubicin in vivo. The MTT assay was used to determine the cytotoxicity of doxorubicin against cancer cells, and apoptosis was analyzed by using flow cytometry. 4T1 cells (2 × 104 cells/mouse) were injected to BALB/c mice, and mice were fed with/without gavage B. infantis milk (108CFU/mL) for 14days and treated with doxorubicin on 5th and 10th days. The weights of the mice were recorded during the study, and the tumor sizes were measured by caliper at the 14th day. CD8 + T cell response was analyzed by using flow cytometer, and the results were compared to control and tumor control groups. The IC50 value for doxorubicin on 4T1 cell lines was determined as 0.053 ± 0.012µg/mL. The apoptotic effect of doxorubicin at IC50 concentration was determined as 82.3% of cells to late apoptosis, 3.6% of cells to pro-apoptosis, and 6.2% of cells to necrosis. The treatment of doxorubicin, B. infantis milk, and the combination of them inhibited the tumor volumes by 55.50%, 40.69%, and 75.95%, respectively. B. infantis administration significantly enhanced the PHA-induced splenocyte proliferation (P < 0.05). It was shown that IFN-γ was effective in tumor growth and regression of metastasis. Consequently, the combination of B. infantis milk and doxorubicin showed the best anti-tumor effect.

Involvement of Mitochondrial Mechanisms and Cyclooxygenase-2 Activation in the Effect of Desethylamiodarone on 4T1 Triple-Negative Breast Cancer Line.

Novel compounds significantly interfering with the mitochondrial energy production may have therapeutic value in triple-negative breast cancer (TNBC). This criterion is clearly fulfilled by desethylamiodarone (DEA), which is a major metabolite of amiodarone, a widely used antiarrhythmic drug, since the DEA previously demonstrated anti-neoplastic, anti-metastasizing, and direct mitochondrial effects in B16F10 melanoma cells. Additionally, the more than fifty years of clinical experience with amiodarone should answer most of the safety concerns about DEA. Accordingly, in the present study, we investigated DEA’s potential in TNBC by using a TN and a hormone receptor positive (HR+) BC cell line. DEA reduced the viability, colony formation, and invasive growth of the 4T1 cell line and led to a higher extent of the MCF-7 cell line. It lowered mitochondrial transmembrane potential and induced mitochondrial fragmentation. On the other hand, DEA failed to significantly affect various parameters of the cellular energy metabolism as determined by a Seahorse live cell respirometer. Cyclooxygenase 2 (COX-2), which was upregulated by DEA in the TNBC cell line only, accounted for most of 4T1’s DEA resistance, which was counteracted by the selective COX-2 inhibitor celecoxib. All these data indicate that DEA may have potentiality in the therapy of TNBC.

Synthesis of doxorubicin‐loaded PBMA‐b‐POEGMA micelles and assessment of its anticancer activity against breast cancer cells (4T1)

AbstractDoxorubicin (DOX) is a chemotherapy drug that possesses many side effects. This study aimed to synthesize PBMA‐b‐POEGMA [poly (butyl methacrylate)‐b‐poly (oligo ethylene glycol methacrylate)] diblock copolymer as a nanocarrier for loading of DOX, promoting anticancer efficacy of the drug, and decreasing its side effects. Hence, PBMA‐b‐POEGMA diblock copolymer was first synthesized by the RAFT method and then DOX encapsulated into the micellar nanocarrier. Self‐assembly behavior of copolymers and physicochemical/biological properties of nanocarriers were assessed. DLS and TEM images showed nanocarriers possessed spherical‐uniform structure with the mean size and polydispersity of 27 ± 1.34 nm and 0.13 ± 0.21 for blank‐micelles as well as, 45 ± 2.32 nm and 0.18 ± 0.18 for DOX‐loaded micelles, respectively. Synthesized micelles also provided high drug encapsulation efficiency (&gt;80%) with a drug loading content of 4.53%. Moreover, the maximum released amount of drug was reported at 69%, with the Korsmeyer‐Peppas model, as a more suitable model to describe the release behavior of DOX from nanocarriers. Biologically, block copolymers were biocompatible against COS‐7 and 4T1 cell lines, in addition, free‐DOX showed higher cytotoxicity than DOX‐loaded micelles. Furthermore, 0.5 μg/mL concentration of drug‐loaded micelle led to growth inhibition of more than 60% of cancerous cells, during 48 h, so that higher level of cellular uptake of drug and apoptosis in 4T1 cells was induced by drug‐loaded micelles.

Monoclonal Antibody Against Sortilin Induces Apoptosis in Human Breast Cancer Cells.

Background:Sortilin has an important role in various malignances and can be used as a promising target to eradicate cancer cells.Methods:In this study, the expression of sortilin in 4T1 and MDA-MB231 cell lines was evaluated by flow cytometry and immunocytochemistry. Apoptosis assay was also applied to evaluate apoptosis induction in 4T1 and MDA-MB231 cell lines.Results:Based on cell surface flow cytometry results, anti-sortilin (2D8-E3) mAb could recognize sortilin molecules in 79.2% and 90.3% of 4T1 and MDA-MB231 cell-lines, respectively. The immunocytochemistry staining results confirmed sortilin surface expression. Apoptosis assay indicated that anti-sortilin mAb could induce apoptosis in 4T1 and MDA-MB231 cell lines.Conclusion:Our study revealed the important role of surface sortilin in breast carcinoma cell survival and its possible application as a therapeutic agent in cancer targeted therapies.

Cytotoxicity and apoptosis assay of novel cyclomyrsinol diterpenes against breast cancer cell lines

Background: Cyclomyrsinane diterpenes especially those extracted from various Euphorbia species have shown interesting biological properties in recent years. Because of the high prevalence of breast cancer and the challenges ahead in its treatment, the use of these compounds as potential anti cancer agents seem reasonable. Objectives: The aim of the present study was to evaluate the cytotoxic effects of some myrsinane type diterpenoids extracted from Euphorbia sogdiana Popov and determine their induced cell death mechanism. Methods: MTT assay was used to determine the cytotoxicity of six various myrsinane compounds on MCF 7 and 4T1 breast cancer cell lines. Human umbilical vein endothelial cells were used as the normal cell line too. The apoptotic effects of the structure with the most cytotoxic effects were determined using flow cytometry assay in IC50 concentration for 24 h of incubation. Results: Compound (6) showed the most cytotoxic effects with IC50 of about 8 ± 4 and 24 ± 4 μg/mL for MCF 7 and 4T1 cell lines, respectively. Furthermore, the cells treated with 5 and 10 μg/mL of compound (6) for 24 h, showed 37 and 55% of apoptotic cells. Conclusions: These surveyed compounds have the potential to be considered as useful anti-breast cancer agents due to the great cytotoxicity and apoptotic effects against these cancer cells and the fact that there was no significant cytotoxicity on normal cells.

Curcumin Sensitizes 4T1 Murine Breast Cancer Cells to Cisplatin Through PAR4 Secretion.

Prostate apoptosis response 4 (PAR4), a tumour-suppressor protein, selectively induces apoptosis of cancer cells without affecting normal cells. Its soluble form is induced by secretagogues (e.g., chloroquine), and it induces apoptosis by interacting with the receptor of glucose-regulated protein 78, which is overexpressed in cancer cells. In this study, curcumin was analyzed as an inducer of PAR4 expression in 4T1 murine breast cancer cell. and its ability to induce PAR4 secretion in Balb/c mice. In addition, the cisplatin sensitizing effect of soluble PAR4 was analyzed. The 4T1 cell line was treated in vitro using different concentrations of curcumin; cell viability was analyzed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and PAR4 expression by western blotting. The expression of soluble PAR4 in the serum of mice treated with intraperitoneal curcumin was analyzed using the dot-blot method. Moreover, MTT assay was used to analyze the effects of serum from curcumin-treated mice on cell viability. Tumor size was analyzed in mice treated with curcumin alone and in combination with cisplatin. Curcumin showed a dose- and time-dependent effects on cell viability on 4T1 cells, as well as increasing PAR4 expression. Compared with the control group (phosphate-buffered saline), mice treated with curcumin showed an increase in plasma PAR4. In the Balb/C tumor model, mice treated with curcumin and cisplatin showed greater tumor shrinkage than the control group. These results indicate that curcumin induces expression of soluble PAR4 and sensitizes tumor cells to cisplatin.

Molecular Imaging of Tumor Progression and Angiogenesis by Dual Bioluminescence.

Angiogenesis is a prerequisite for tumor growth and invasion, and anti-angiogenesis has become a highlight in tumor treatment research. However, so far, there is no reliable solution for how to simultaneously visualize the relationship between tumor progression and angiogenesis. Bioluminescence imaging (BLI) has been broadly utilized and is a very promising non-invasive imaging technique with the advantages of low cost, high sensitivity, and robust specificity. In this chapter, we describe a dual bioluminescence imaging BLI protocol for tumor progression and angiogenesis through implanting murine breast cancer cell line 4T1 which stably expressing Renilla luciferase (RLuc) into the transgenic mice with angiogenesis-induced firefly luciferase (FLuc) expression. This modality enables us to synchronously monitor the tumor progression and angiogenesis in the same mouse, which has broad applicability in oncology studies.

Protective effect of Di'ao Xinxuekang capsule against doxorubicin-induced chronic cardiotoxicity

Ethnopharmacological relevanceDi'ao Xinxuekang capsule (DXXK) extracted from Dioscorea nipponica Makino is a well-known traditional Chinese herbal medicinal product widely used in the treatment of cardiovascular disease, such as myocardial ischemia and arrhythmia. The active ingredients of DXXK were also traditionally utilized for treating cardiovascular disease in the former Soviet Union after the 1960s. As a specific type of cardiovascular disease, doxorubicin (DOX)-induced cardiotoxicity is characterized by arrhythmia, myocardial ischemia, and heart failure. Aim of the studyThis study aimed to investigate the potential protective effect of DXXK against chronic cardiotoxicity induced by DOX. Materials and methodsA mouse model of chronic cardiotoxicity induced by DOX and an in vitro model of DOX-induced myocardial damage were created to assess the protective effect of DXXK. Cardiac functional parameters, serum levels of CK-MB and LDH and cardiac histopathological indicators were determined in the mouse model. Moreover, cell viability was measured by the MTT method, and the effect of DXXK on the anticancer activity of DOX was also investigated by utilizing 4T1, HepG2, and H460 cell lines. Furthermore, the levels of markers of oxidative stress indexes (SOD, GSH, MDA) and inflammation (TNF-α, IL-1α) were measured using biochemical and Elisa kits, respectively. The level of ROS in H9c2 cardiomyocyte was determined by flow cytometry. The protein expression levels of HIF-1α and NF-κB p65 were measured by western blotting. Finally, molecular docking was performed to visualize the patterns of interactions between the effective molecule and targeted protein. ResultsDXXK alleviated DOX-induced chronic cardiotoxicity as shown by the reversal of changes in levels of myocardial enzymes and left ventricular function and structure. DXXK exhibits antioxidant and anti-inflammatory activities. We also observed that DXXK might increase the protein expression level of HIF-1α and decrease the protein expression level of NF-κB p65. Further results of in vitro experiments showed that DXXK could protect cardiomyocyte against DOX-induced production of ROS, but DXXK had no effect on the anticancer activity of DOX. The results of molecular docking showed that dioscin and pseudoprotodioscin were the top two compounds of DXXK, which had high affinity with HIF-1α and NF-κB p65. ConclusionsOur results indicated that DXXK could protect against cardiotoxicity induced by DOX and alleviate oxidative stress and inflammation in vivo and in vitro via the regulation of HIF-1α and down NF-κB p65.

Iron Oxychloride/Bovine Serum Albumin Nanosheets as Chemodynamic Therapy Agents

AbstractIron oxychloride (FeOCl) is known for reactive oxygen species (ROS) generation through Fenton chemistry. The activity of FeOCl is preserved in the slightly acidic pH value of the tumor microenvironment (pH 6.5−6.9). Such property can be advantageous in biobased systems, where ROS generation can be modulated in slightly acidic conditions, which is characteristic of the solid tumor microenvironment. In the present study, BSA‐stabilized FeOCl nanosheets (NSs) are synthesized and characterized by transmission electron microscope, Fourier transform infrared spectroscopy, zeta potential analysis, dynamic light scattering, and UV–vis spectroscopy. The morphology of the nanoparticles is flake‐like, and their hydrodynamic diameter is around 200 nm. MTT, apoptosis assay, and trypan blue staining evaluate the toxicity of FeOCl NSs toward the 4T1 cell line. It is found that the toxicity of the NSs is higher in physiological conditions of solid tumors (pH 6.5, H2O2 100 × 10−6 m) than in the conditions of healthy organs (pH 7.4). Specifically, cancer cells are in their late apoptotic stage by more than eight times higher at pH 6.5 than pH 7.4. The toxicity results are in agreement with the in vitro catalytic assay of the NSs. Therefore, the FeOCl NSs can be the building blocks for constructing chemodynamic therapy agents.

Yerba Mate Modulates Tumor Cells Functions Involved in Metastasis in Breast Cancer Models.

Breast cancer (BC) is the most frequent cancer in women and tumor metastasis is a major cause of cancer-related deaths. Our aim was to evaluate anti-metastatic properties of yerba mate extract (YMe) in BC models. 4T1, F3II, MCF-7, and MDA-MB231 cell lines were used to perform in vitro assays. The F3II syngeneic mammary carcinoma model in BALB/c mice was used to evaluate tumor progression, BC metastasis and survival. Cells were inoculated subcutaneously into the flank for the heterotopic model and into the mammary fat pad for the orthotopic model. YMe was administered p.o. in a dose of 1.6 g/kg/day. In vitro YMe inhibited cell proliferation and reduced tumor cell adhesion, migration and invasion. These biological effects were cell-line dependent. In vivo YMe reduced tumor metastasis and increased mice survival in both models. Our preclinical results suggest that YMe could modulate tumor progression and metastasis in BC models.

Isolation and Characterization of Crab Haemolymph Exosomes and Its Effects on Breast Cancer Cells (4T1).

Objective The use of animal or plant exosomes in cancer treatment is promising because of their easy access and low cost. Freshwater crabs are used in traditional Iranian medicine to treat cancer. This study aims to determine the anti-cancer properties of exosomes removed from freshwater crabs on a breast cancer cell line (4T1) compared to bone marrow mesenchymal stem cells (BMSCs). Materials and Methods In this experimental study, crab haemolymph exosomes were isolated via the precipitation method and characterised by electron microscopy, dynamic light scattering (DLS), and Western blot analysis. The protein concentration and total antioxidant capacity of these exosomes were determined by bicinchoninic acid (BCA) and cupric reducing antioxidant capacity (CUPRAC). The 4T1 cells and BMSCs were treated with exosomes and we assessed the cell survival by the resazurin and MTT assays. The level of nitric oxide (NO) secretion from the 4T1 cells was determined after treatment with the exosomes. Results Electron microscopy, DLS and Western blot for CD63 confirmed that the isolated exosomes were <100 nm in size and expressed CD63. The total antioxidant capacity in these exosomes was 1.003 µM/ml and the protein concentration was 650 mg/ml. Resazurin and MTT assay results showed a decrease in survival of the 4T1 cells (P≤0.001) after treatment with the exosomes compared to cell growth in the exosome-treated BMSCs. ConclusionCrab haemolymph contains protein-rich exosomes with antioxidant activities that can have anti-cancer effects on 4T1 cells. These exosomes may be proposed for breast cancer therapeutics.

In vitro anti-proliferative effect of capecitabine (Xeloda) combined with mocetinostat (MGCD0103) in 4T1 breast cancer cell line by immunoblotting.

Objective(s):Mouse breast cancer cell line 4T1 can accurately mimic the response to immune receptors and targeting therapeutic agents. Combined therapy has emerged as an important strategy with reduced side effects and maximum therapeutic effect. Mocetinostat (MGCD0103) is one of the members of Class I Histone Deacetylase Inhibitors (HDACi) and its mechanism of action has not been defined, yet. Capecitabine (Xeloda) is an antimetabolite and currently is widely utilized to treat a wide range of solid tumors. The aim of this study was to investigate the effects of the capecitabine, mocetinostat and their combined application on the 4T1 cell line. Materials and Methods:The effects of combined administration of mocetinostat and capecitabine on 4T1 cells were investigated by cell viability and migration assays, apoptosis analysis, and Western blotting technique.Results:The concentrations of drugs that give a half-maximal response (IC50) were detected for capecitabine (1700 µM), mocetinostat (3,125 µM), and 50 µM Capecitabine+1,5 µM Mocetinostat for 48 hr. In capecitabine+mocetinostat combine group, we observed that cell migration decreased, DNA fragmentation increased compared to the control group. capecitabine + mocetinostat group induced apoptosis by decreasing Bcl-2, PI3K, Akt, c-myc protein levels, while increasing Bax, Caspase-3, PTEN, cleaved-PARP, Caspase-7, Caspase-9, p53, cleaved-Cas-9 protein levels in 4T1 cells. Conclusion:Capecitabine and mocetinostat played a toxic role through inducing apoptosis on 4T1 cancer cells in a time- and concentration-dependent manner. These results showed that combined therapy with low concentrations were detected to be more effective than that with high-concentration alone drug treatment.

Annonaceous acetogenins nanosuspensions stabilized by poloxamer 188: Preparation, properties and in vivo evaluation

Annonaceous acetogenins (ACGs) have extensive antitumor activities. However, the poor solubility limits their clinical application. In this research, ACGs nanosuspensions (ACGs-NSps) were prepared by antisolvent-precipitation method using poloxamer 188 (P188) as stabilizer. The prepared P188-ACGs-NSps had a mean particle size of 115.9 nm, the polydispersity index (PDI) was 0.208. P188-ACGs-NSPs were stable in different physiological media and displayed a sustained drug release behavior until 144 h.Lyophilized P188-ACGs-NSps using 0.5% glucose as lyoprotectant were stable for long-term storage. P188-ACGs-NSps significantly enhanced the cytotoxicity against 4T1 and HeLa cell lines compared with ACGs solution (IC50:1.426 ± 0.308 vs.3.106 ± 0.691, IC50: 0.718 ± 0.1934 vs. 1.521 ± 0.1992). In vivo antitumor study indicated that P188-ACGs-NSps exhibited great antitumor efficacy against 4T1 tumor with an inhibition rate of 63% at a low intravenous injection dose of 0.4 mg/kg. P188-ACGs-NSps seem to be a promising drug delivery system for ACGs to improve their solubility that can be applied in clinic in the future.

Steroidal glycosides from the Vietnamese cultivar Cordyline fruticosa “Fairchild red”

A phytochemical study of Cordyline fruticosa “Fairchild red” (Asparagaceae) from Vietnam, led to the isolation of fourteen steroidal glycosides, including twelve previously undescribed along with two known ones. Ten compounds were obtained by successive solid/liquid chromatographic methods from an aqueous-ethanolic extract of the roots, and four from the aerial parts. Their structures were elucidated mainly by spectroscopic analysis 2D NMR and mass spectroscopy (ESI-MS), as spirostanol glycosides, 5α-spirost-25(27)-ene-1β,3β,4α-triol 1-O-β-D-fucopyranoside, 5α-spirost-(25)27-ene-1β,3β,4α-triol 1-O-β-D-xylopyranoside, 5α-spirost-(25)27-ene-1β,3β,4α-triol 1-O-α-L-rhamnopyranosyl-(1 → 2)-β-D-fucopyranoside, 5α-spirost-(25)27-ene-1β,3β,4α-triol 1-O-α-L-rhamnopyranosyl-(1 → 2)-(4-O-sulfo)-β-D-fucopyranoside, 5α-spirost-25(27)-ene-1β,3β-diol 1-O-α-L-rhamnopyranosyl-(1 → 2)-β-D-fucopyranoside, and 5α-spirost-25(27)-ene-1β,3β-diol 1-O-α-L-rhamnopyranosyl-(1 → 2)-α-L-arabinopyranoside. Furostanol glycosides were also isolated as 26-O-β-D-glucopyranosyl-5α-furost-(25)27-ene-1β,3β,4α,22α,26-pentol 1-O-β-D-fucopyranoside, 26-O-β-D-glucopyranosyl-22α-methoxy-5α-furost-(25)27-ene-1β,3β,4α,26-tetrol 1-O-β-D-fucopyranoside, 26-O-β-D-glucopyranosyl-5α-furost-(25)27-ene-1β,3β,22α,26-tetrol 1-O-β-D-glucopyranoside, 26-O-β-D-glucopyranosyl-5α-furost-(25)27-ene-1β,3β,22α,26-tetrol 1-O-α-L-rhamnopyranosyl-(1 → 2)-β-D-glucopyranoside, 26-O-β-D-glucopyranosyl-5α-furost-(25)27-ene-1β,3β,22α,26-tetrol 1-O-α-L-rhamnopyranosyl-(1 → 2)-β-D-fucopyranoside, and 26-O-β-D-glucopyranosyl-22α-methoxy-5α-furost-(25)27-ene-1β,3β,26-triol 1-O-α-L-rhamnopyranosyl-(1 → 2)-β-D-fucopyranoside. All the isolated compounds were further evaluated for their cytotoxicity against 4T1 cell line, from a mouse mammary gland tissue, using MTS method.

Selectivity of Ethanol Extract of Parijoto (Medinilla speciosa) Fruit in HepG2, Widr, 4T1, and Vero cell lines

Background: Parijoto, one of the melastomaceae family, has been known to have cytotoxic activity in some cancer cell lines, such as HeLa, MCF-7, and T47D. Aims: We aim to know about the selectivity of ethanol extract of Parijoto fruit in cell line HepG2, WiDr, 4T1, and Vero. Cytotoxic was determined by MTT assay. Method: Extract was added in three serial concentration three serial concentrations (125 µg/mL–500 µg/mL), while the positive control doxorubicin gives in 2,5 µg/mL – 20 µg/mL for cancer cell and 40 µg/mL -100 µg/mL for Vero cell. Results: Results showed that ethanol extract of parijoto fruit gave low activity in HepG2 and Vero cell (IC50: 250 µg/mL) and moderate activity in WiDr and 4T1 (IC50: 81,58 µg/mL and 158,72 µg/mL). Conclusion: The highest selectivity index is given in WiDr cell (SI&gt; 3) means that the ethanol extract of parijoto fruit is a promising cytotoxic agent for colorectal cancer therapy.

Correlation Between Antioxidant and Cytotoxic Activity of Parijoto (Medinilla speciosa Blume) Fractions in 4T1 Cell Line

Parijoto (Medinilla speciosa Blume) is one of Indonesian plant used for traditional medicine. Previous studies have demonstrated antimicrobial and cytotoxic effects of Parijoto on T47D cells. Therefore, we intended to know the antioxidant and cytotoxic activity of these fractions in 4T1 cell line (a Mus musculus mammary carcinoma). This cancer causes the greatest number of cancer-related deaths This study also investigated the correlation between antioxidant activity and cytotoxicity of Parijoto fractions. Discovering the type of correlation between antioxidant and anticancer activity of botanical extracts could relieve in screening for cytotoxic agent from natural products. The antioxidant and cytotoxic activity investigated using the Diphenylpicrylhydrazyl (DPPH) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay methods. The result showed that ethyl acetate fraction is the higher antioxidant activity (IC50:1.77 μg/mL) and the higher cytotoxicity (IC50:133.57 μg/mL). There was a strong positive correlation (correlation coefficient=0.957) between antioxidant and cytotoxic activity in 4T1 cell line, but the correlation was not significant (p=0.188).Keywords: Parijoto (Medinilla speciosa Blume), antioxidant, cytotoxic, 4T1 cell line.

Pentagamaboronon-0-Sorbitol Induces Apoptosis through Elevation of Reactive Oxygen Species in Triple Negative Breast Cancer Cells

Breast cancer is the most common type of cancer causing mortality for women due to metastasis. More than 50% of breast cancer patients are suffered lung metastases. One strategy to target the cancerous cell is Boron Neutron Captured Therapy (BNCT) which showed high affinity toward cancer cells and reported to have anti-proliferative as well as anti-metastatic activities. Pentagamaboronon-0 (PGB-0) is a curcumin analogue substance which had reported to exert anticancer activities against Her-2 expressing as well as triple negative breast cancer cells. Despite its great potency as BNCT agent candidate, this compound also exerted several anticancer properties. Complex formation of this substance with sorbitol was achieved to improve the solubility and maximize compound’s delivery to the target cells. This study aimed to investigate the ability of Pentagamaboronon-0-Sorbitol (PGB-0-So) to modulate cell cycle and induce apoptosis especially through the mechanisms of reactive oxygen species (ROS) modulation. The 3-(4,5-dimethylthiazzol-2yl)-2,5-diphenyltetrazolium (MTT) cytotoxicity assay of PGB-0-So against 4T1 breast cancer cell line were found to exert potential effect in dose-dependent manner with lethal concentration (IC50) values of 39 μM. The cytotoxicity of PGB-0-So complex was found to be increased considerably compared with that of PGB-0. Cell cycle modulation identified using propidium iodide (PI) staining showed cell accumulation in S phase following treatment with PGB-0-So. Apoptosis induction assay analyzed using flowcytometer with Annexin V and PI staining on its IC50 dose was found to induce programmed cell death (apoptosis). The sub-IC50 treatment of this compound was also improved the cellular ROS level which also took role in apoptosis induction. These findings suggest that PGB-0-So is potential as an anticancer agent.Keywords: Curcumin analogue, PGB-0-So, Anticancer, 4T1 cell line, ROS modulation.

Curcumin-like structure (CCA-1.1) induces permanent mitotic arrest (Senescence) on Triple-negative breast cancer (TNBC) cells, 4T1

Triple-negative breast cancer (TNBC) remains as the deadliest cancer type due to the lack of treatment options. Hence, several attempts have been made to develop new anticancer for TNBC therapy. This study intended to challenge curcumin analog (CCA)-1.1, which is derived from pentagamavunone-1 structure, against the 4T1 cell line and TNBC cell model, covering the cytotoxic activity in correlation with cell cycle progression, apoptosis induction, reactive oxygen species (ROS) generation, and senescence evidence. The cell viability, cell cycle profile, apoptosis induction, intracellular ROS level, and senescence induction were determined in vitro using trypan blue exclusion, propidium iodide (PI) staining, Annexin-PI staining, dichlorofluorescein diacetate staining, and senescence-associated-β-gal method. CCA-1.1 showed cytotoxic activity on 4T1 cells, giving half maximal inhibitory concentration value of 3M, but was less toxic on non-cancerous 3T3-L1 cells. CCA-1.1 induced rapid cell death and inhibited cell cycle progression at the mitotic phase. Instead, of causing apoptosis, CCA-1.1 induced mitotic catastrophe. Furthermore, CCA-1.1 itself increased the intracellular ROS level and induced senescence, possibly through catastrophic cell death. Altogether, our preliminary study strengthens the potency of CCA-1.1 for its anticancer activities against TNBC cells and prospective to be pharmaceutically developed as a novel candidate for cancer therapy.