Xanthorrhizol Induces Apoptosis Via the Up-regulation of Bax and p53 in HeLa Cells

Xanthorrhizol is a sesquiterpenoid compound extracted from Curcuma xanthorrhiza, which is known locally as Temulawak. Traditionally, C. xanthorrhiza was found to have antibacterial, anticancer and anti-inflammatory activity. The rhizome has also been used to treat inflammation in postpartum uterine bleeding. An antiproliferative assay using methylene blue staining revealed that xanthorrhizol inhibited the proliferation of the cervical cancer cell line HeLa with an EC50 value of 6.16 Ìg/ml. Xanthorrhizol significantly increased apoptosis in HeLa cells, as evaluated by the Tdt-mediated dUTP nick end-labelling (TUNEL) assay and nuclear morphology by Hoechst 33258 staining. Western blot analysis, which was further confirmed by the immunostaining results, implied an up-regulation of tumor suppressor protein p53 and the pro-apoptotic protein Bax, following the treatment with xanthorrhizol. Xanthorrhizol, however, did not affect the expression of the anti-apoptotic protein, Bcl-2 and the viral oncoprotein, E6. Hence, xanthorrhizol is a promising antiproliferative and anticancer agent which induces p53 and Bax-dependent apoptosis in HeLa cervical cancer cells

Xanthorrhizol Induces Apoptosis Via the Up-regulation of Bax and p53 in HeLa Cells

Xanthorrhizol is a sesquiterpenoid compound extracted from Curcuma xanthorrhiza, which is known locally as Temulawak. Traditionally, C. xanthorrhiza was found to have antibacterial, anticancer and anti-inflammatory activity. The rhizome has also been used to treat inflammation in postpartum uterine bleeding. An antiproliferative assay using methylene blue staining revealed that xanthorrhizol inhibited the proliferation of the cervical cancer cell line HeLa with an EC50 value of 6.16 Ìg/ml. Xanthorrhizol significantly increased apoptosis in HeLa cells, as evaluated by the Tdt-mediated dUTP nick end-labelling (TUNEL) assay and nuclear morphology by Hoechst 33258 staining. Western blot analysis, which was further confirmed by the immunostaining results, implied an up-regulation of tumor suppressor protein p53 and the pro-apoptotic protein Bax, following the treatment with xanthorrhizol. Xanthorrhizol, however, did not affect the expression of the anti-apoptotic protein, Bcl-2 and the viral oncoprotein, E6. Hence, xanthorrhizol is a promising antiproliferative and anticancer agent which induces p53 and Bax-dependent apoptosis in HeLa cervical cancer cell

Natural luteolin from methanolic extract of Malaysian brucea javanica leaves induces apoptosis in hela cell lines

There are a growing number of deaths of cancer patients due to toxicity of conventional chemotherapy. Therefore, continuous discovery of natural anticancer compounds with cytoselective actions would be an ideal strategy to overcome the problem. Thus, natural products from plant are still the alternative source in the search for anticancer drugs that can have a direct effect on both cancer cell removal and also minimize the side effects to the patients. Based on the traditional usage and pharmaceutical potential of Brucea javanica reported before, a study on the leaves of this plant was carried out. The aims of the study were to isolate the bioactive compound from B. javanica Leaves (BJL) extracts via bioassay-guided fractionation using several selected cancer cell lines, to determine the mode of cancer cell death induced by BJL’s active compound and the molecular mechanism of apoptosis implicated in cancer cell lines by measuring the level of apoptotic protein expression such as bax, bcl-XL, caspase-3 and tumour suppressor p53. Among three crude extracts of BJL, methanol was the most potent against selected cancer cell lines which consist of cervical, breast, bone, ovarian and liver cancer cells. Cisplatin was used as positive control for the antiproliferative assay. Using a bioassay-guided fractionation, chromatography, NMR and mass spectrometry analysis we have isolated luteolin. It is a known compound from the flavonoid group which was found to be cytoselective. The IC50 value for HeLa is 8.02 μg/ml while for Vero is >99 μg/ml. Hoechst 33258 staining and flow cytometric analysis of Annexin V-FITC staining revealed that luteolin induces apoptotic cell death in cervicalcancer cell HeLa. Flow cytometric analysis also showed that luteolin induces apoptosis by increasing the p53, bax and caspase-3 protein expression

GC-MS Analysis of Phytochemical Compounds in Aqueous Leaf Extract of Abrus Precatorius

Abrus precatorius is a flowering plant that belongs to the legume family, Fabaceae. In Malaysia, the leaves of Abrus precatorius are used traditionally to treat ailments such as fever, ulcer and mouth cancer. These traditional practices, however, have never been documented and usage of the plant is based on popular beliefs held by the local people. This work documented the phytocompounds that are present in the aqueous extract of Abrus precatorius leaves collected from a local area in Kota Bharu, Kelantan, Malaysia. The leaves were dried and then subjected to extraction using the decoction technique. The compounds were identified by gas chromatography with mass spectrometry analysis and characterised by comparison through the NIST02 and Wiley275 library search software. The GC-MS analysis showed that the classes of compounds identified in aqueous extracts of Abrus precatorius leaves were phenolic compounds, terpenoids and steroid

Human natural killer (NK) cell activation by luteolin from Brucea javanica leaves

There is growing interest in using natural substances from plants to potentially enhance anticancer activity. A key target in this respect is natural killer (NK) cells in order to generate an anticancerimmune response. Luteolin used in this study was derived from a local plant Brucea javanica leaves extract; through a bioassay-guided fractionation. The expression of interleukin-2 (IL-2) and interferon-γ (IFN-γ) was increased after 20 h treatment of 0.1 µM luteolin in human mixed lymphocytes, which reflects the activation of NK cells. This was further confirmed in co-culture experiments. NK cellinduced K562 target cell death was increased in the presence of luteolin. These results show that luteolin activates NK cells to kill target cells indicating the potential use of luteolin as an anticancer immunostimulant

Pereskia bleo leaves extract induces cell death via cell cycle arrest and apoptosis in cervical cancer cells hela

Introduction: Pereskia bleo is a leafy and edible plant, locally known as “Pokok Jarum Tujuh Bilah” which has anticancer properties. This study purposed to determine the cytotoxic effects of P. bleo leaves extracts on several well-known cancer cells and elucidate its underlying mechanism in inducing cell death. Methods: Cytotoxic activity on selected cell lines was determined using MTT assay. Mechanism of cell death was investigated through cell cycle and Annexin V assay. Expression of apoptotic proteins was measured by flow cytometry method. Results: Ethyl acetate extract of P. bleo leaves (PBEA) appeared to have the strongest IC50 value (14.37 ± 8.40 lg/ml) and most active against HeLa cells was further studied for apoptosis. The cell cycle investigation by flow cytometry evidenced the increment of PBEA treated HeLa cells in G0/G1 phase and apoptotic event was detected in Annexin V assay. Analysis of apoptotic protein showed pro-apoptotic proteins (Bax, p53 and caspase 3) were triggered where as anti-apoptotic protein Bcl-2 was suppressed in treated HeLa cells. Conclusions: Our findings demonstrated that PBEA treatment induced cell death in HeLa cells by p53-mediated mechanism through arresting cell cycle at G0/G1 phase and mitochondrial-mediated pathway with involvement of pro-apoptotic proteins, anti-apoptotic protein, and caspase 3

Phytochemical Screening and Cytotoxic Effects of Crude Extracts of Pereskia Bleo Leaves

The phytocompounds in crude solvent extracts of Pereskia bleo leaves were identified and their cytotoxic effects on cancer cell lines were determined. Crude extracts were obtained via maceration and subjected to GCMS analysis. Then, each extract was incubated with HeLa, MDA-MB-231, SW480, and NIH/3T3 cell lines for 72 h. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was done to determine IC50 values of each extract. Terpenoids, sterols, alkaloids, fatty acids and phenolic compounds were identified from the crude extracts of P. bleo leaves. Other compounds identified were γ-sitosterol, β-tocopherol, and γ-tocopherol. The ethyl acetate extract had potent cytotoxic effect against HeLa and MDA-MB-231 cancer cells as noted by the lowest IC50 value

Unveiling safety profile of Pereskia bleo leaf extract: A comprehensive study on female toxicity and teratogenicity in Sprague Dawley rats

Pereskia bleo (P. bleo), renowned for its traditional medicinal use in various countries, is valued for its therapeutic potential. While medicinal plants hold immense promise, concerns regarding potential adverse effects on normal cells and teratogenicity remain. This study evaluated the influence of methanol extract of Pereskia bleo (MEPB) leaves on female toxicity and teratogenicity in Sprague Dawley rats. Female rats (n=40) were divided into four groups receiving MEPB doses (0, 250, 500, and 1000 mg/kg/day) during pre-mating, mating, and gestation periods. Gas Chromatography-Mass Spectrometry (GC-MS) analysis exhibited that MEPB leaves contain phenolic compounds, terpenoids, phytosterols, and fatty acids. Remarkably, all the animals exhibited regular oestrous cycles, maintaining body weight, and showed no signs of toxicity or abnormal behavior. Visceral organ weights and histological analysis revealed no significant alterations, attesting to the extract's safety. Pregnancy outcomes, including corpora lutea, implantation sites, percentages of pre- and post- implantation death, gravid uterine weight, number of live and dead fetuses, fetal body weight, and fetal sex ratio, remained unaffected. Furthermore, meticulous fetal examinations confirmed that MEPB did not impact foetal parameters or induce deformities. Importantly, daily MEPB consumption (250-1000 mg/kg) from pre-pregnancy to pregnancy did not compromise maternal or fetal well-being. These results underscore the lack of embryotoxic or teratogenic effects of MEPB, highlighting its potential for safe therapeutic applications

Methanolic extract of Abrus precatorius promotes breast cancer MDA-MB-231 cell death by inducing cell cycle arrest at G0/G1 and upregulating Bax

Objective: To determine the anti-proliferative activity of Abrus precatorius (A. precatorius) leaf extracts and their effect on cell death. Methods: A. precatorius leaves were extracted successively with hexane, ethyl acetate and methanol by Soxhlet extraction. Aqueous extract was prepared by decoction at 50. Extracts of A. precatorius leaves were used to treat selected cancer and normal cell lines for 72 h. Furthermore, 3-(4,5-dimethyl thiazol-2-yl) 2,5-diphenyl tetrazolium bromide assay was performed to determine cell viability. Analysis of cell cycle arrest, apoptosis assay and apoptosis protein expressions were determined by flow cytometry. Results: Methanolic extract of A. precatorius leaves showed the lowest IC50 on MDA-MB-231 cells at (26.40依5.40) μg/mL. Flow cytometry analysis revealed that cell arrest occurred at G0/ G1 phase and the apoptosis assay showed the occurrence of early apoptosis at 48 h in MDAMB- 231 cells treated with methanolic extract of A. precatorius leaves. Methanolic extract of A. precatorius leaves induced apoptosis by upregulation of Bax, p53 and caspase-3 and downregulation of Bcl-2. Conclusions: Methanolic extract of A. precatorius leaves promotes MDA-MB-231 cell death by inducing cell cycle arrest and apoptosis possibly via the mitochondrial-related pathway

Gold nanoparticles conjugated with anti-CD133 monoclonal antibody and 5-fluorouracil chemotherapeutic agent as nanocarriers for cancer cell targeting

The enhanced permeability and retention effect allows for passive targeting of solid tumours by nanoparticles carrying anticancer drugs. However, active targeting by incorporation of various ligands onto nanoparticles can provide for a more selective and enhanced chemotherapeutic effect and complement the deficiencies of the passive targeting approach. Here we report on the design of the carboxyl-terminated PEGylated gold nanoparticles (AuNPs), their functionalization with anti-CD133 monoclonal antibody (mAb) via a crosslinking reaction, and subsequent 5-fluorouracil (5-FU) drug loading. The synthesized products in the form of stable colloids were characterised using a range of physicochemical techniques, including X-ray diffraction (XRD), UV-Vis spectroscopy, transmission electron microscopy (TEM), and dynamic light scattering (DLS). Conjugation of anti-CD133 mAb onto PEGylated AuNPs was confirmed with the use of UV-Vis, BCA protein assay and fluorescence microscopy. HCT116 colorectal cancer cells abundantly expressed CD133: 92.4 ± 1.3%, as measured by flow cytometry. Whereas PEGylated AuNPs not conjugated with anti-CD133 mAb accumulated mainly at the cellular membrane, nanoparticles conjugated with anti-CD133 mAb were contained within the nuclear region of the cells. Anti-CD133 mAb conjugation facilitated the specific intracellular uptake due to specific antigen–antibody binding interaction. In vitro cytotoxicity studies on HCT116 cells showed that PEGylated AuNPs and PEGylated AuNPs-CD133 did not elicit any toxicity at any of the tested concentrations. Meanwhile, 5-FU-PEGylated AuNPs-CD133 significantly reduced the cell viability relative to the treatment with 5-FU-PEGylated AuNPs without anti-CD133 mAb conjugates (p < 0.0001). This study shows that the conjugation of nanocarriers with the anti-CD133 antibody improves the specific targeting of 5-FU against colorectal cancer cells. These results demonstrate that simultaneous functionalisation of PEGylated AuNPs with antibodies and chemotherapeutic drugs is a viable strategy to combat cancer through targeted drug delivery