Human Cell Line PC-3 Research Articles

Identification of placenta-specific protein 1 (PLAC-1) expression on human PC-3 cell line-derived prostate cancer stem cells compared to the tumor parental cells

Placenta-specific protein 1 (PLAC-1) is a gene primarily expressed in the placenta and the testis. Interestingly, it is also found to be expressed in many solid tumors, and it is involved in malignant cell features. However, no evidence has been reported regarding the relationship between PLAC-1 and cancer stem cells (CSCs). In the current research, we explored the expression of the PLAC-1 molecule in prostate cancer stem cells (PCSCs) derived from the human PC-3 cell line. The enrichment of PCSCs was achieved using a three-dimensional cell culture technique known as the sphere-formation assay. To confirm the identity of PCSCs, we examined the expression of genes associated with stemness and pluripotency, such as SOX2, OCT4, Nanog, C-Myc, and KLF-4, as well as stem cell differentiation molecules like CD44 and CD133. These evaluations were conducted in both the PCSCs and the original tumor cells (parental cells) using real-time PCR and flow cytometry. Subsequently, we assessed the expression of the PLAC-1 molecule in both enriched cells and parental tumor cells at the gene and protein levels using the same techniques. The tumor cells from the PC-3 cell line formed spheroids with CSC characteristics in a non-adherent medium. The expression of SOX2, OCT4, Nanog, and C-Myc genes (p < 0.01), and the molecules CD44 and CD133 (p < 0.05) were significantly elevated in PCSCs compared to the parental cells. The expression of the PLAC-1 molecule in PCSCs showed a significant increase compared to the parental cells at both gene (p < 0.01) and protein (p < 0.001) levels. In conclusion, it was indicated for the first time that PLAC-1 is up-regulated in PCSCs derived from human PC-3 cell line. This study may propose PLAC-1 as a potential target in targeted therapies, which should be confirmed through further studies.

Novel photothermal therapy using multi-walled carbon nanotubes and platinum nanocomposite for human prostate cancer PC3 cell line

Multi-walled carbon nanotubes (MWCNTs) coated with platinum nanoparticles (Pt-NPs) was prepared as a novel composite nanomaterial by using waste synthetic oil and green chemistry. MWCNT and Pt-NPs were characterised through TEM, FE-SEM, AFM and FT-IR techniques. The diameters of the synthesised MWCNTs and Pt-NPs were 26.80–44.66 and 56.49–89.38 nm, respectively. The formation of MWCNT-COOPt and MWCNT-Pt was confirmed using Raman and EDX techniques. The effect of MWCNT-COO, MWCNT-COOPt and MWCNT-Pt on prostate cancer cell line PC3 was studied by MTT assay at various concentrations following near-infrared (NIR) irradiation (λ=1064 nm, P = 15.3 W) at different time intervals (30, 60, 90 and 120 s). The composite of Pt-NPs upon the MWCNT surface enhanced its ability to absorb NIR radiation, leading to an increase in the temperature of cancer cells due to plasmon phenomenon. The composites were utilised in a novel treatment against human PC3 cell line. The maximum temperatures for MWCNT-COO, MWCNT-COOPt and MWCNT-Pt recorded with 25 µg/mL were 43.4 °C, 45.8 °C and 46.2 °C, respectively, and irradiation time was recorded at 120 s. These compounds exhibited high cytotoxicity towards human PC3 cells (58.6%, 71.6% and 79.6%), respectively. The combination of MWCNTs and Pt-NPs in photothermal therapy has potential to be used in local therapy for prostate cancer in a time- and concentration-dependent manner.

Reduction of oxidative stress and ornithine decarboxylase expression in a human prostate cancer cell line PC-3 by a combined treatment with α-tocopherol and naringenin.

Differentiation of a human aggressive PC-3 cancer cell line was obtained, in a previous investigation, by the synergic effect of α-tocopherol (α-TOC) and naringenin (NG). This combined treatment induced apoptosis and subsequent reduction of the PC-3 cell proliferation and invasion, by a pro-differentiating action. Since one of the peculiar characteristics of NG and α-TOC is their strong antioxidant activity, this study aimed to investigate their potential effect on the activity of the main enzymes involved in the antioxidant mechanism in prostate cancer cells. NG and α-TOC administered singularly or combined in the PC-3 cell line, affected the activity of several enzymes biomarkers of the cellular antioxidant activity, as well as the concentration of total glutathione (GSH + GSSG) and thiobarbituric acid reactive substances (TBARS). The combined treatment increased the TBARS levels and superoxide dismutase (SOD) activity, while decreased the glutathione S-transferase (GST), glutathione reductase (GR), and glyoxalase I (GI) activities. The results obtained indicate that a combined treatment with these natural compounds mitigated the oxidative stress in the human PC-3 cell line. In addition, a significant reduction of both ornithine decarboxylase (ODC) expression and intracellular levels of polyamines, both well-known positive regulators of cell proliferation, accompanied the reduction of oxidative stress observed in the combined α-TOC and NG treatment. Considering the established role of polyamines in cell differentiation, the synergism with NG makes α-TOC a potential drug for further study on the differentiation therapy in prostate cancer patients.

PEGylated Carbon Nanotubes Decorated with Silver Nanoparticles: Fabrication, Cell Cytotoxicity and Application in Photo Thermal Therapy

A new technique for cancer therapy is Photo Thermal Therapy (PTT). In the PTT technique, photon energy is converted into heat via various operations to destroy malignant tumors. Carbon nanotubes (CNTs) have good optical absorption in the near-infrared (NIR) spectrum and could transform optical energy into heat to induce hyperthermia in the PTT method. In this study, CNTs were firstly oxidized (O-CNT) and then decorated with silver nanoparticles (Ag NPs). Polyethylene glycol (PEG) was utilized for wrapping the surface of CNTs (O-CNT/Ag-PEG). Coating of CNTs with Ag NPs and PEG was confirmed by XRD, FESEM, and TEM techniques. Results demonstrated that noble metal could increase optical absorption of CNTs and concurrently improve the efficacy of the PTT technique. Cell cytotoxicity study showed that O-CNT/Ag NPs were less cytotoxic than O-CNTs, and O-CNT/Ag-PEG had the lowest toxicity against HeLa, HepG2, and PC3 human cell lines. The efficacy of O-CNT/Ag-PEG NPs in destroying malignant melanoma tumors was evaluated through the PTT technique. A continuous wave NIR laser diode (λ = 808 nm, P = 2 W, and I = 2 W/cm2) irradiated the tumor sites for 8 min once in the period of the treatment. The tumors in cases receiving O-CNT/Ag-PEG were shrunk efficiently compared to laser treatment ones. Results of in-vivo studies demonstrated that O-CNT/Ag-PEG was a puissant candidate in extirpating malignant tumors in PTT method.

Effect of Lutein on Proliferation and Apoptosis of Human Prostate Cancer PC3 Cell Line and Related Mechanism

Effect of Lutein on Proliferation and Apoptosis of Human Prostate Cancer PC3 Cell Line and Related Mechanism

Antitumor activity of L-asparaginase produced from E.coli .

.Extra and intra-cellular L-asparaginase were produced from E.coli, purified by ammonium sulphate precipitation, ion exchange chromatography and gel filtration chromatography and tested for their anti-tumor activities against four human tumor cell lines include MCF-7 , PC3 , A549 and Hep-G2 using MTT and (HCS) array scan. The MTT assay showed that the enzymes exhibited antiproliferative activity in different cell lines growth, PC3 cell line were more sensitive to extra and intracellular L-asparaginase as it gave a lower IC50 value (27.41±2.95 and 70.12±2.53) for extra and intracellular respectively, extracellular L-asparaginase treatment increases nuclear condensation, plasma membrane permeability, MMP attenuation and increased cytochrome C in the cystol compared to control. We suggest the extra-cellular Lasparaginase of E.coli was a therapeutic agent against human PC3 cell line.

Cytotoxic cardiac glycosides from the fruit (pods) of Adenium obesum (Forssk.) Roem. & Schult

Phytochemical investigation of methanolic extract of Adenium obesum led to the isolation of 42 (1–42) compounds belongs to cardiac glycosides, triterpenoids and steroids. The chemical structures of isolated compounds were elucidated by spectral techniques UV, IR, NMR and FAB MS. The cardiac glycosides were tested against three human cell lines, 3T3 (normal cells), HeLa (Human cervical cancer cell lines) and PC-3 (Human prostate cancer cell lines). The cardiac glycoside, honghelin (4), obeside B (5) and obeside C (6) showed significant effects against cell lines Hela, 3T3 and PC-3 compared to standard drug doxorubicin. Compounds 4, 5 and 6 exhibited very low IC50 (μM) against the PC3 human cell line. 4 and 6 also showed least IC50 against the HeLa human cell lines as compared to the standard drug doxorubicin whereas these three compounds showed effect on 3T3 cell line with high IC50 values compared to drug cycloheximide.

Abstract B38: Photochemical internalisation for the local delivery of chemotherapy in prostate cancer

Abstract • Introduction Prostate cancer is fifth leading cause of death from cancer in men. Although chemotherapy is the gold standard treatment for a wide range of cancers including the prostate, there are several limitations (systemic toxicity, multidrug resistance or inadequate cellular delivery) and there is growing interest in developing focal therapies when appropriate. Photochemical Internalisation (PCI) is a focal light-triggered drug delivery system, which combines low dose Photodynamic Therapy (PDT) with chemotherapy agents to enhance delivery of the agents to their specific subcellular target. PCI is particularly suitable for agents prone to endolysosomal sequestration, thererefore the photosensitisers (PS) used for PCI localise in endolysosomal membranes which upon visible light exposure leads to the production of reactive oxygen species (ROS), causing photooxidative disruption of membranes and subsequent cytosolic release of the entrapped agents. These will then reach their intended targets more efficiently, improving efficacy and potentially reducing the therapeutic dose. The present project focuses on the use of PCI to deliver Saporin, a ribosome inactivating (RIP) type 1 protein. RIPs are able to cause permanent damage to eukaryotic ribosomes; however, despite its intrinsic high toxicity, endolysosomal sequestration impairs its ability to inhibit protein synthesis. • Aim To assess the potential of PCI for enhanced delivery of Saporin in human and rat prostate cancer cells. In addition, a CXCR4-targeted peptide-PS conjugate was tested. • Materials &amp; Methods Porphyrin and chlorin-based PS were co-administered with Saporin. Cells were incubated for 24hrs with different drug combinations then exposed to light, and cell viability was measured at different time points post illumination using the MTT assay. Subcellular location of Saporin and PS were observed using confocal microscopy. • Results PCI resulted in a synergistic effect between sub-lethal PDT and Saporin. Despite minimal cell kill when the toxin or PS were administered alone, PCI delivery enhanced Saporin cytotoxicity using only 20pM in the rat Mat-LyLu and 2nM in PC3 human cell lines. PCI elicits up to 1.5-fold higher efficacy in cell kill vs. PDT 48hrs after light treatment and 3.5-fold higher 96hrs after light treatment in rat and human prostate cancer cells accordingly (p&amp;lt;0.05). In addition, PS doses were reduced by 20 times when targeted against CXCR4 receptor (overexpressed in prostate cancer among others). This resulted in 75% reduction of cell viability compared to 79% using the 20-fold higher dose of untargeted PS. Moreover, a 3-fold higher efficacy in cell kill of PCI vs. PDT 96hrs after light treatment in human cells was observed. Intracellular redistribution following endolysosomal release of fluorescently labelled Saporin and PS following illumination was confirmed with confocal microscopy. • Conclusions These results demonstrate the potential of PCI for prostate cancer treatment in vitro and in vivo tumour experiments are in progress. Citation Format: ALEJANDRA MARTINEZ DE PINILLOS, RUGGERO DONDI, L. WANG, ELNAZ YAGHINI, JOSEPHINE H. WOODHAMS, CAROLINE M. MOORE, IAN M. EGGLESTON, ALEXANDER J. MACROBERT. Photochemical internalisation for the local delivery of chemotherapy in prostate cancer. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr B38.

Phytofabrication of biomolecule-coated metallic silver nanoparticles using leaf extracts of in vitro-raised bamboo species and its anticancer activity against human PC3 cell lines

The present study describes the biosynthesis of metallic silver nanoparticles (AgNPs) from silver ions using in vitro-grown leaf samples of Bambusa arundinacea (Ba) and Bambusa nutans (Bn) and their anticancer activity against human prostatic cancer cell lines (PC3). The metallic AgNPs were synthesized at room temperature by treating the leaf extracts of Bambusa arundinacea and Bambusa nutans with 2 mM silver nitrate using the boiled method. The characterization of synthesized BaAgNPs and BnAgNPs was performed using UV-Vis spectrum (absorption peak at 365 nm), FTIR, TEM (spherical shape), SEM (size of 30-90 nm), and EDX analysis. Cell viability assays were carried out to determine the cytotoxicity effect of AgNPs on the PC3 cancer cell line and the Vero normal cell line. The IC_50 values for BaAgNP- and BnAgNP-treated PC3 cancer cells were found to be 73.57 µg/mL and 84.88 µg/mL while the values for treated Vero cells were found to be 93.58 µg/mL and 96.41 µg/mL, respectively. The percentages of the apoptotic bodies by AO/EtBr staining were found to be 76% and 62% for BaAgNPs and BnAgNPs, respectively. The results strongly suggest that synthesized BaAgNPs show potential anticancer activity against human PC3 cell lines compared to BnAgNPs.

Microwave-mediated extracellular synthesis of metallic silver and zinc oxide nanoparticles using macro-algae (Gracilaria edulis) extracts and its anticancer activity against human PC3 cell lines.

A rapid and novel microwave-mediated protocol was established for extracellular synthesis of metallic silver (Ag) and zinc oxide (ZnO) nanoparticles using the extracts of macro-algae Gracilaria edulis (GE) and also examined its anticancer activity against human prostate cancer cell lines (PC3). The formation of silver nanoparticles (GEAgNPs) and zinc oxide nanoparticles (GEZnONPs) in the reaction mixture was determined by ultraviolet-visible spectroscopy. The synthesized Ag and ZnO nanoparticles were characterized by X-ray diffraction, Fourier transform infra-red spectroscopy, energy dispersive X-ray, and field emission scanning electron microscopy. The silver and zinc oxide nanoparticles were spherical and rod-shaped, respectively. Cell viability assays were carried out to determine the cytotoxic effects of AgNPs and ZnONPs against PC3 and normal African monkey kidney (VERO) cell line. The inhibitory concentration values were found to be 39.60, 28.55, 53.99 μg/mL and 68.49, 88.05, 71.98 μg/mL against PC3 cells and Vero cells for AgNPs, ZnONPs, and aqueous G. edulis extracts, respectively, at 48 h incubation period. As evidenced by acridine orange/ethidium bromide staining, the percentage of the apoptotic bodies was found to be 62 and 70 % for AgNPs and ZnONPs, respectively. The present results strongly suggest that the synthesized ZnONPs showed an effective anticancer activity against PC3 cell lines than AgNPs.

Evaluation of N-[(11)C]methyl-AMD3465 as a PET tracer for imaging of CXCR4 receptor expression in a C6 glioma tumor model.

The chemokine receptor CXCR4 and its ligand CXCL12 play an important role in tumor progression and metastasis. CXCR4 receptors are expressed by many cancer types and provide a potential target for treatment. Noninvasive detection of CXCR4 may aid diagnosis and improve therapy selection. It has been demonstrated in preclinical studies that positron emission tomography (PET) with a radiolabeled small molecule could enable noninvasive monitoring of CXCR4 expression. Here, we prepared N-[(11)C]methyl-AMD3465 as a new PET tracer for CXCR4. N-[(11)C]Methyl-AMD3465 was readily prepared by N-methylation with [(11)C]CH3OTf. The tracer was obtained in a 60 ± 2% yield (decay corrected), the purity of the tracer was >99%, and specific activity was 47 ± 14 GBq/μmol. Tracer stability was tested in vitro using liver microsomes and rat plasma; excellent stability was observed. The tracer was evaluated in rat C6 glioma and human PC-3 cell lines. In vitro cellular uptake of N-[(11)C]methyl-AMD3465 was receptor mediated. The effect of transition metal ions (Cu(2+), Ni(2+), and Zn(2+)) on cellular binding was examined in C6 cells, and the presence of these ions increased the cellular binding of the tracer 9-, 7-, and 3-fold, respectively. Ex vivo biodistribution and PET imaging of N-[(11)C]methyl-AMD3465 were performed in rats with C6 tumor xenografts. Both PET and biodistribution studies demonstrated specific accumulation of the tracer in the tumor (SUV 0.6 ± 0.2) and other CXCR4 expressing organs, such as lymph node (1.5 ± 0.2), liver (8.9 ± 1.0), bone marrow (1.0 ± 0.3), and spleen (1.0 ± 0.1). Tumor uptake was significantly reduced (66%, p < 0.01) after pretreatment with Plerixafor (AMD3100). Biodistribution data indicates a tumor-to-muscle ratio of 7.85 and tumor-to-plasma ratio of 1.14, at 60 min after tracer injection. Our data demonstrated that N-[(11)C]methyl-AMD3465 is capable of detecting physiologic CXCR4 expression in tumors and other CXCR4 expressing tissues. These results warrant further evaluation of N-[(11)C]methyl-AMD3465 as a potential PET tracer for CXCR4 receptor imaging.

WE-G-BRE-08: Radiosensitization by Olaparib Eluting Nanospheres

Purpose: Permanent prostate brachytherapy often uses inert bio-absorbable spacers to achieve the desired geometric distribution of sources within the prostate. Transforming these spacers into implantable nanoplatforms for chemo-radiation therapy (INCeRT) provides a means of providing sustained in-situ release of radiosensitizers in the prostate to enhance the therapeutic ratio of the procedure. Olaparib, a PARP inhibitor, suppresses DNA repair processes present during low dose rate continuous irradiation. This work investigates the radiosensitizing/DNA damage repair inhibition by NanoOlaparib eluting nanospheres. Methods: Human cell line PC3 (from ATCC), was maintained in F12-k medium supplemented with fetal bovine serum. Clonogenic assay kit (from Fischer Scientific) was used to fix and stain the cells to determine the long term effects of irradiation. Nanoparticle size and zeta potential of nanospheres were determined using a Zeta particle size analyzer. The incorporation of Olaparib in nanospheres was evaluated by HPLC. Irradiation was performed in a small animal irradiator operating at 220 KeV. The long term effects of radio-sensitization with olaparib and nanoolaparib was determined using the clonogenic assay at 2 Gy and 4 Gy doses. The cells were allowed to grow for around 10 doubling cycles, The colonies were fixed and stained using clonogenic assay kit. The excess stain was washed off using DI water and the images were taken using a digital camera. Results: Radiosensitization studies were carried out in prostate cancer cell line, PC3 radiation at 0, 2 and 4Gy doses. Strongest dose response was observed with nanoolaparib treated cells compared to untreated cells. Conclusion: A two stage drug release of drug eluting nanospheres from a biodegradable spacer has been suggested for sustained in-situ release of Olaparib to suppress DNA repair processes during prostate brachytherapy. The Olaparib eluting nanospheres had the same in-vitro radiosensitizing effect as free olaparib. DOD 1R21CA16977501, A. David Mazzone Awards Program 2012PD164

Comprehensive analysis of human cells motion under an irrotational AC electric field in an electro-microfluidic chip.

AC electrokinetics is a versatile tool for contact-less manipulation or characterization of cells and has been widely used for separation based on genotype translation to electrical phenotypes. Cells responses to an AC electric field result in a complex combination of electrokinetic phenomena, mainly dielectrophoresis and electrohydrodynamic forces. Human cells behaviors to AC electrokinetics remain unclear over a large frequency spectrum as illustrated by the self-rotation effect observed recently. We here report and analyze human cells behaviors in different conditions of medium conductivity, electric field frequency and magnitude. We also observe the self-rotation of human cells, in the absence of a rotational electric field. Based on an analytical competitive model of electrokinetic forces, we propose an explanation of the cell self-rotation. These experimental results, coupled with our model, lead to the exploitation of the cell behaviors to measure the intrinsic dielectric properties of JURKAT, HEK and PC3 human cell lines.

Prevalence of Prostate Cancer Metastases after Intravenous Inoculation Provides Clues into the Molecular Basis of Dormancy in the Bone Marrow Microenvironment

Bone is the preferred metastasis site of advanced prostate cancer (PCa). Using an in vivo murine model of human PCa cell metastasis to bone, we noted that the majority of animals that develop skeletal metastasis have either spinal lesions or lesions in the bones of the hindlimb. Much less frequently, lesions develop in the bones of the forelimb. We therefore speculated whether the environment of the forelimb bones is not permissive for the growth of PCa. Consequently, data on tumor prevalence were normalized to account for the number of PCa cells arriving after intravascular injection, marrow cellularity, and number of hematopoietic stem cell niches. None of these factors were able to account for the observed differences in tumor prevalence. An analysis of differential gene and protein levels identified that growth arrest specific-6 (GAS6) levels were significantly greater in the forelimb versus hindlimb bone marrow. When murine RM1 cells were implanted into subcutaneous spaces in immune competent animals, tumor growth in the GAS6-/- animals was greater than in GAS6+/+ wild-type animals. In an osseous environment, the human PC3 cell line grew significantly better in vertebral body transplants (vossicles) derived from GAS6-/- animals than in vossicles derived from GAS6+/+ animals. Together, these data suggest that the differences in tumor prevalence after intravascular inoculation are a useful model to study the molecular basis of tumor dormancy. Importantly, these data suggest that therapeutic manipulation of GAS6 levels may prove useful as a therapy for metastatic disease.

Synthesis and structure–activity relationship study of novel cytotoxic carbamate and N-acylheterocyclic bearing derivatives of betulin and betulinic acid

Chemical transformation studies were conducted on betulin and betulinic acid, common plant-derived lupane-type triterpenes. The concise synthesis, via a stepwise approach, of betulin and betulinic acid carbamate and N-acylheterocyclic containing derivatives is described. All new compounds, as well as betulinic acid were tested in vitro for their cytotoxic activity. Most of the compounds have shown a better cytotoxic profile than betulinic acid, including the synthesized betulin derivatives. Compounds 25 and 32 were the most promising derivatives, being up to 12-fold more potent than betulinic acid against human PC-3 cell lines (IC 50 values of 1.1 and 1.8 μM, respectively).