Prostate Carcinoma Cell Lines Research Articles

Cytotoxicity of AMANTADIG – a semisynthetic digitoxigenin derivative – alone and in combination with docetaxel in human hormone-refractory prostate cancer cells and its effect on Na+/K+-ATPase inhibition

Cardiac glycosides (CGs) are natural compounds used to treat congestive heart failure. They have garnered attention as a potential cancer treatment option, especially because they bind to Na+/K+-ATPase as a target and activate intracellular signaling pathways leading to a variety of cellular responses. In this study we evaluated AMANTADIG, a semisynthetic cardenolide derivative, for its cytotoxic activity in two human androgen-insensitive prostate carcinoma cell lines, and the potential synergistic effects with docetaxel. AMANTADIG induced cytotoxic effects in both cell lines, and a combination with docetaxel showed a moderate and strong synergism in DU145 and PC-3 cells, respectively, at concentrations considerably lower than their IC50 values. Cell cycle analyses showed that AMANTADIG and its synergistic combination induced G2/M arrest of DU145 and PC-3 cells by modulating Cyclin B1, CDK1, p21 and, mainly, survivin expression, a promising target in cancer therapy. Furthermore, AMANTADIG presented reduced toxicity toward non-cancerous cell type (PBMC), and computational docking studies disclosed high-affinity binding to the Na+/K+-ATPase α subunit, a result that was experimentally confirmed by Na+/K+-ATPase inhibition assays. Hence, AMANTADIG inhibited Na+/K+-ATPase activity in PC-3 cells, as well as in purified pig kidney at nanomolar range. Altogether, these data highlight the potent effects of AMANTADIG in combination with docetaxel and offer important insights for the development of more effective and selective therapies against prostate cancer.

Abstract 3923: Insensitivity to c-Met inhibition at physiologic HGF concentrations in a prostate carcinoma model

Abstract C-Met is a receptor tyrosine kinase critical for embryogenesis and liver repair. In tumors, including breast, prostate, glioblastoma, and thyroid cancer, Met protein levels are often elevated and associated with disease progression and metastasis. The c-Met pathway is activated by the endogenous ligand Hepatocyte Growth Factor (HGF). HGF is produced by mesenchymal cells and binds to the c-Met protein, leading to phosphorylation and upregulation of a variety of downstream signaling pathways that result in increased tumor cell migration, proliferation, survival, and induction of angiogenesis. C-Met is associated with disease progression and metastasis. Consequently, there has been significant interest in the development of c-Met inhibitors as anti-cancer therapeutics. The efficacy of such agents usually initially are determined in vitro utilizing HGF concentrations of 25-50 ng/mL. However, HGF serum levels in humans typically range from 0.4-0.8 ng/mL. In our studies, we examined the prostate carcinoma cell line DU145, and BMS-777607. Migration and phosphorylation of c-Met increased in a dose-dependent manner with HGF treatment. Additionally, c-Met inhibition with BMS-777607 reduced migration and invasion when in the presence of 25-50 ng/mL HGF. However, at physiologically relevant HGF concentrations the drug had no effect on migration or invasion of this cell line. In vivo models of c-Met inhibition have been limited mainly to mice genetically engineered to express HGF, on the basis that mouse HGF does not stimulate human c-Met. However, in our studies, mouse HGF did stimulate c-Met in the DU145 tumor cell line. Subsequent in vivo evaluation revealed treatment with BMS-777607 had no reduction on DU145 tumor cell-induced angiogenesis in cells pretreated with the drug, nor in mice receiving oral administration of the drug. An experimental metastasis model also resulted in no significant reduction in metastasis formation, as assessed by a tail vein injection model. To characterize HGF concentration in vivo, an ELISA was used to determine an average concentration of 7.7 ng/g of DU145 tumor tissue. Further evaluations of phospho-Met are underway utilizing immunofluorescence of FFPE samples. Preliminary results indicate low levels of phospho-Met in DU145 tumor sections that can be induced by treating tumor-bearing mice with exogenous HGF. Our results suggest that in vitro evaluation of c-Met inhibitors utilizing non-physiological concentrations of HGF may produce an abnormally high phospho-Met that can subsequently be reduced by treatment with c-Met inhibition. In vitro studies performed at high HGF concentrations may not accurately predict the in vivo drug efficacy. Further characterization of HGF concentrations within the tumor is necessary to properly conduct in vitro evaluation of c-Met inhibitors. Citation Format: Veronica S. Hughes, Mumtaz Rojiani, Dietmar Siemann. Insensitivity to c-Met inhibition at physiologic HGF concentrations in a prostate carcinoma model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3923.

Design, synthesis and in vitro evaluation of heterobivalent peptidic radioligands targeting both GRP- and VPAC1-Receptors concomitantly overexpressed on various malignancies – Is the concept feasible?

Radiolabeled heterobivalent peptidic ligands (HBPLs), being able to address different receptors, are highly interesting tumor imaging agents as they can offer multiple advantages over monovalent peptide receptor ligands. However, few examples of radiolabeled HBPLs have been described so far. One promising approach is the combination of gastrin-releasing peptide receptor (GRPR)- and vasoactive intestinal peptide receptor subtype 1 (VPAC1R)-targeting peptides into one single radioligand since gastrinomas, prostate and breast cancer have been shown to concomitantly or complementarily overexpress both receptors. Here we report the design and synthesis of different HBPLs, comprising a GRPR-binding (BBN7-14) and a VPAC1R-targeting (PACAP-27) peptide. The heterodimers were varied with regard to the distance between the peptide binders and the steric rigidity of the systems. We radiolabeled the HBPLs 19–23 as well as their monomeric reference standards 26 and 27 with 68Ga, achieving radiochemical yields and purities of 95–99% and non-optimized molar activities of 25–61 GBq/μmol. We tested the stability of the radioligands and further evaluated them in vitro regarding their uptake in different prostate carcinoma cell lines (PC-3, DU-145 and VCaP cells).We found that the heterobivalent substances [68Ga]19 − [68Ga]23 showed comparable uptakes into the tumor cells to those of the respective monomers [68Ga]26 and [68Ga]27, indicating that both peptides are still able to address their target receptors. Furthermore, the obtained results indicate that in case of overall low receptor densities, heterobivalent peptides surpass peptide monomers in tumor cell uptake. Most importantly, it could be shown by blocking studies that both peptide parts of the HBPL [68Ga]19 contributed to tumor cell uptake in VCaP cells, expressing both receptor types.Thus, we describe here the first examples of HBPLs being able to address the GRPR as well as the VPAC1R and have the potential to − by several mechanisms − improve tumor targeting for several malignancies compared to monospecific peptides.

Oxazoline derivatives of [17(20)E]-21-norpregnene – inhibitors of CYP17A1 activity and proliferation of prostate carcinoma cells

Nine new oxazolinyl derivatives of [17(20)E]-21-norpregnene, differing in the structure of steroid moiety, were investigated for their potency to inhibit the CYP17A1 catalytic activity, as well as growth and proliferation of LNCaP and PC-3 prostate carcinoma cell lines. The activity of one of the investigated compounds, 2´-{[(E)3β-hydroxyandrost-5-en-17-ylidene]methyl}-4´,5´-dihydro-1´,3´-oxazole (1), was found to be comparable with that of Abiraterone, a known inhibitor of CYP17A1 used for treatment of prostate cancer. A model of the interaction of oxazoline 1 with the active site of CYP17A1 was constructed by the molecular dynamics method. A correlation was found between the structure and the biological activity in the series of [17(20)E]-21-norpregnene oxazolinyl derivatives.

Major terpenoids from Telekia speciosa flowers and their cytotoxic activity in vitro

In addition to known constituents of Telekia speciosa, an acetone extract from ray florets of the plant yielded: 5,5ʹ-dibutoxy-2,2ʹ-bifuran (1), 5,5ʹ-diisobutoxy-2,2ʹ-bifuran (2), α-tocopherol (3), β-tocopherol (4), loliolide palmitate (5), a mixture of calenduladiol esters - 16β-hydroxylupeol-3-O-palmitate (7) and 16β-hydroxylupeol-3-O-myristate (8), 1-epiinuviscolide (12), inuviscolide (13), 3-epiisotelekin (16), 4α-hydroxy-9β,10β-epoxy-1β(H)-11(13)-guaien-8α,12-olide (17), 4α-hydroxy-1β(H)-9(10),11(13)-guaiadien-8α,12-olide (18), loliolide (19) and 4β,10β-dihydroxy-1α(H),5α(H)-11(13)-guaien-8α,12-olide (20). Calenduladiol esters and asperilin (14) were the major constituents of the extract. Their cytotoxic effect on human normal prostate epithelial cells (PNT-2), human prostate carcinoma cell lines, human skin fibroblasts (HSF) and human melanoma cell lines was examined in vitro. Triterpene esters showed no cytotoxicity against nearly all cell lines tested, except for Du145 prostate carcinoma cells (IC50 – 62.0 μΜ). Asperilin displayed activity against the cell lines under study, especially against three tested lines of melanomas (A375, IC50 – 17.6 μΜ, WM793, IC50 – 28.2 μΜ and Hs 294T, IC50 – 29.5 μΜ).

DTPA-(PABn)-Leu5]-des-acyl ghrelin(1-5) as a new carrier of radionuclides and potential precursor of radiopharmaceuticals.

Ghrelin is a peptide consisting of 28 aminoacids and an octadecyl side chain (acyl group) binding to the growth hormone secretagogue receptor type 1a (GHS-R1a). Its des-acylated form, des-acyl ghrelin (DAG) binds to the corticotropin releasing factor receptor type 2a (CRF2a) located on endocrine cancer cells such as the prostate carcinoma cell line DU 145. The aim of this study is to develop a new DAG-based carrier of radionuclides with potential application in therapy. Trunctated C-terminal five aminoacids chain of the DAG peptide (H2N-Gly-Ser-Ser-Phe-Leu-COOH) was linked to DTPA to obtain [DTPA-(PABn)-Leu5]-DAG(1-5). For therapeutic application the lutetium-177 (177Lu) radionuclide was coordinated to the peptide. To determine biological and chemical properties of newly synthesized radiopharmaceutical, two iodine-131 (131I)-labelled compounds were used: [131I]-Tyr4-DAG(1-5) and full length [131I]-DAG(1-28) together with their nonradioactive forms: DAG(1-28) and DAG(1-5). Identical HPLC elution profiles of [177Lu-DTPA-(PABn)-Leu5]-DAG(1-5) before and after incubation with human serum proved its stability. The lipophilicity profile of [177Lu-DTPA-(PABn)-Leu5]-DAG(1-5) was log DO/W=-2.68±0.05, pH 7.4. Receptor affinity of the nonradioactive conjugate [Lu-DTPA-(PABn)-Leu5]-DAG(1-5) was IC50 (21.06 nmol/l), as shown against the [131I]-DAG(1-28) used as a competitor. The 3-(4,5-dimethyldiazol-2-yl)-2,5-diphenyltetrazolium bromide assay indicated the significant cytotoxicity of the newly synthesized compounds, similar to that of [131I]-Tyr4-DAG(1-5). The results obtained suggest the potency of the [DTPA-(PABn)-Leu5]-DAG(1-5) as a new carrier of radionuclides in radiopharmacy.

Potential anticancer activity of biogenic silver nanoparticles using leaf extract of Rhynchosia suaveolens: an insight into the mechanism

The present study reports a simple and eco-friendly synthesis of silver nanoparticles (AgNPs) using leaf extract of Rhynchosia suaveolens. UV-Vis analysis of R. suaveolens synthesized AgNPs (RS-AgNPs) showed surface plasmon resonance (SPR) peak at 426 nm. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) analysis revealed that RS-AgNPs were 10–30 nm in size with spherical shape. X-ray diffraction (XRD) analysis of RS-AgNPs confirmed the crystalline nature with face-centered cubic (FCC) lattice. Fourier transform infrared (FTIR) interprets that polyphenols and proteins take part in bioreduction and capping of RS-AgNPs. RS-AgNPs exhibited dose-dependent inhibition of proliferation of different cancer cells including DU145 and PC-3(human prostate carcinoma cell lines), SKOV3 (human ovarian carcinoma) and A549 (human lung adenocarcinoma)with IC50 values of 4.35, 7.72, 4.2 and 24.7 μg/mL, respectively. The plausible reasons behind anticancer activity of RS-AgNPs were explained using different assays on the most susceptible SKOV3 cells. RS-AgNPs induced oxidative stress in SKOV3 cells by generating reactive oxygen species (ROS), enhancing lipid peroxidation (LPO) levels and decreasing glutathione (GSH) levels. RS-AgNPs induced the apoptosis of SKOV3 cells by up regulating the caspase-3, caspase -8, caspase -9, p53 and BAX and down regulating the antiapoptotic protein Bcl-2. Further, RS-AgNPs showed elevation of caspase 3/7 activity and also exhibited antimigratory effect by inhibiting the migration of SKOV3 cells into the wounded area. The findings suggested that biogenic RS-AgNPs provide an alternative approach to overcome several limitations of chemotherapy.

Synthesis of Nano Sulfur Particles and their Antitumor Activity

Objectives: The present study is designed to evaluate the antitumor activity of sulphur containing compounds in Nano formulations against Ehrlisch Ascites Carcinoma (EAC) in female albino mice. Methods: Preparation of sulfur nanoparticles (S-NPs) was done throughout. Firstly, the evaluation of their composition and purity by fourier transform infrared spectrometer (FT-IR) analysis. Secondly, the shape and size of S-NPs were investigated by scanning electron microscopy (SEM) techniques in all S-NPs samples which were prepared with Tetramethylammoniumbromide (TMAB) surfactant, Then, Energy-dispersive X-ray spectroscopy (EDS) for the evaluation of their composition and purity, after that the X-Ray diffraction patterns were measured for prepared S-NPs with TMAB surfactant, and the determination of S-NPs antitumor activity against (breast carcinoma cell line (MCF7), liver carcinoma cell line (HEPG2), colon carcinoma cell line (HCT116), prostatic carcinoma cell line (PC3). Finally, determination biochemical assay of S-NPs by using female mice bearing EAC and of the assessment of liver functions and evaluation of kidney functions and determination of some heart functions in serum and hematology markers. Results: This study revealed that S-NPs showed high strong activities in concentration (5 mg/kg) as it improvement of liver functions production compared to the positive control group, also showed the effect of S-NPs of the kidney functions, heart functions and hematology parameters in all studied groups and a significant increase in activities compared with positive control group. Conclusion: S-NPs play an important role in improving liver functions, kidney functions and the heart functions of resulted in significantly increase activity against tumour and plays an important role in improving oxidative stress resulted in high antiproliferation activity against (MCF-7, HepG2, HCT116 and PC3) cell lines, and significantly increase with S-NPs reduced most of the hematological parameters towards the best compared to the positive control group.

Cytotoxic Effects of Rhododendron ponticum L. Extract on Prostate Carcinoma and Adenocarcinoma Cell Line (DU145, PC3)

Cytotoxic Effects of Rhododendron ponticum L. Extract on Prostate Carcinoma and Adenocarcinoma Cell Line (DU145, PC3)

Flavonolignans and Biological Activity of Nannorrhops ritchiana Leaves

Objectives: This study aimed at isolating the polyphenolic compounds of 70% methanol extract ofNannorrhops ritchiana Griff. leaves and assaying the antioxidant and cytotoxic activities of the extract and main fractions. Methods: The methanol extract of the leaves of N. ritchiana was chromatographically fractionated using a bioactivity-guided approach. The isolated compounds were spectroscopically elucidated by UV, MS, 1H/13C NMR and 2D NMR spectroscopic techniques. The radical scavenging activity of the methanol extract and its fractions was evaluated using DPPH (2,2-Diphenyl-1-picrylhydrazyl) radical scavenging assay and their cytotoxic activity was assayed using SRB (3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyltetrazolium bromide) test against human hepatocellular liver carcinoma (HepG2), human alveolar adenocarcinoma (A549) and human prostate carcinoma (PC3) cell lines. Results: Eight compounds were isolated and identified for the first time from the leaves of N. ritchiana, comprising five flavonolignans;Indocalatin A (4) which is a rare compound that has been reported only once in nature, 9″-O-glucopyranosyl Salcolin A (5), 9″-O-glucopyranosyl Salcolin B (6) together with their aglycones, Salcolin A (7) and Salcolin B (8) along with three flavone glycosides; Tricin-7-O-rutinoside (1), Orientin (2) and Isoorientin (3). The methanol extract of N. ritchiana showed antioxidant activity with SC50 39.4 ± 1.06 μg/ml, while fraction IIshowed significant effect with SC50 of 6.1 ± 0.24 μg/ml in comparison with ascorbic acid (SC50 1.8 ± 0.35 μg/ml). The methanol extract and the fraction III showed a promising cytotoxic activity against selected cell lines especially the A549 with IC50 9.5 ± 1.98 μg/ml and HepG2 with IC50 8.15 ± 1.3 μg/ml, respectively. Conclusion: N. ritchiana leaf methanol extract is a new source of biologically active compounds, including flavonolignans.

Cyclin-dependent kinase inhibitor 3 (CDKN3) plays a critical role in prostate cancer via regulating cell cycle and DNA replication signaling

Cyclin-dependent kinase inhibitor 3 (CDKN3) is proved to be associated with the progressing of many cancers. Whereas, its biological effects on prostate cancer (PC) are less understood. To investigate the functional mechanism of CDKN3 in PC, we examined the expression of CDKN3 in PC tissues and analyzed the disease free survival time of patients. We then transfected LNCaP and PC3 cells with siRNA-CDKN3 to silence CDKN3, and transfected 22RV1 and VCaP cells with full length CDKN3 cDNA for CDKN3 over-expression. Cell growth of these transfected cells were analyzed using CCK-8 assay. And transfected LNCaP and PC3 cells were further submitted to cell cycle, apoptosis, invasion and endogenous protein expression assays. We found that CDKN3 was highly expressed in PC and negatively correlated with disease relapse. And CDKN3 positively control the cell proliferation in prostate carcinoma cell lines. Knockdown of CDKN3 significantly promoted G1 phase arrest, elevated apoptosis rates, and suppressed cell invasion in both LNCaP and PC3 cells. Moreover, in vivo data showed that knockdown of CDKN3 expression dramatically inhibited the PC3 tumor growth in nude mouse model. Gene set enrichment analysis (GSEA) showed that cell cycle and DNA replication signaling were related with elevated CDKN3 expression. And results of western blot showed that the depletion of CDKN3 down-regulated the expression levels of cell cycle- and DNA replication-related proteins. In conclusion, our results highlight the importance of CDKN3 in PC and provide new insights into diagnostics and therapeutics of the PC.

Coatomer subunit beta 2 (COPB2), identified by label-free quantitative proteomics, regulates cell proliferation and apoptosis in human prostate carcinoma cells

Label-free quantitative proteomics has broad applications in the identification of differentially expressed proteins. Here, we applied this method to identify differentially expressed proteins (such as coatomer subunit beta 2 [COPB2]) and evaluated the functions and molecular mechanisms of these proteins in prostate cancer (PCA) cell proliferation. Proteins extracted from surgically resected PCA tissues and adjacent tissues of 3 patients were analyzed by label-free quantitative proteomics. The target protein was confirmed by bioinformatics and GEO dataset analyses. To investigate the role of the target protein in PCA, we used lentivirus-mediated small-interfering RNA (siRNA) to knockdown protein expression in the prostate carcinoma cell line, CWR22RV1 cells and assessed gene and protein expression by reverse transcription quantitative polymerase chain reaction and western blotting. CCK8 and colony formation assays were conducted to evaluate cell proliferation. Cell cycle distributions and apoptosis were assayed by flow cytometry. We selected the differentiation-related protein COPB2 as our target protein based on the results of label-free quantitative proteomics. High expression of COPB2 was found in PCA tissue and was related to poor overall survival based on a public dataset. Cell proliferation was significantly inhibited in COPB2-knockdown CWR22RV1 cells, as demonstrated by CCK8 and colony formation assays. Additionally, the apoptosis rate and percentage of cells in the G1 phase were increased in COPB2-knockdown cells compared with those in control cells. CDK2, CDK4, and cyclin D1 were downregulated, whereas p21 Waf1/Cip1 and p27 Kip1 were upregulated, affecting the cell cycle signaling pathway. COPB2 significantly promoted CWR22RV1 cell proliferation through the cell cycle signaling pathway. Thus, silencing of COPB2 may have therapeutic applications in PCA.

Insight on cytotoxic effects of silver nanoparticles: Alternative androgenic transactivation by adsorption with DHT

Silver nanoparticles (AgNPs) are accumulated in the male reproductive organs for a long time and cause several adverse effects in there. Up to now, there is little of information for the cytotoxic effects in male reproductive cells. In this study, the stable AgNPs with a minimal silver ion (Ag+) dissolution below concentration inducing cytotoxicity in the cell medium were exposed to the human prostate carcinoma cell line 22Rv1. Moreover particle uptake and androgen receptor (AR) transactivation were evaluated. In cell medium, AgNPs exhibited stability in an aqueous environment and minimal Ag+ release. Transmission electron microscopy (TEM) and energy dispersive spectrometer (EDS) analysis demonstrated uptake of AgNPs into cells via endocytosis, and a quantitative Ag assay showed that uptake of AgNPs was size-dependent with the majority of Ag retained in the particle form. To evaluate if the presence of AgNPs can change androgenic potentials of dihydrotestosterone (DHT, strong human androgen), we conducted an AR transactivation assay using the transgenic prostate cell line 22Rv1-MMTV-Hyg and found that AgNPs lowered androgenic transactivation of DHT, which is due to decreased bioavailability of DHT.

Effect of Levisticum officinale Hydroalcoholic Extract on DU-145 and PC-3 Prostate Cancer Cell Lines

: Levisticum officinale contains a variety of components used for its medicinal properties in traditional medicine. The previous studies showed that it had an anti-cancer effect on cell lines. The current study aims to evaluate the cytotoxic potential of Levisticum officinale hydroalcoholic extract (LOHE) on DU-145 and PC-3 human advanced prostate cancer cell lines. The anti-proliferative activity of LOHE was evaluated using the MTT assay. The results of MTT showed that the IC50 of DU-145 and PC-3 were 500 µg/mL and 400 µg/mL of LOHE, respectively, following 24 hours treatment. This study found that the hydroalcoholic extract of Levisticum officinale had beneficial cell death inducing effects on both the PC3 and DU145 human prostate carcinoma cell lines in time- and concentration-dependent fashion. Hence, LOHE may have remarkable potential for uses as new natural anticancer agent.

Label-Free Photoelectrochemical "Off-On" Platform Coupled with G-Wire-Enhanced Strategy for Highly Sensitive MicroRNA Sensing in Cancer Cells.

MicroRNA (miRNAs) quantification, especially at low abundance, is vital for disease diagnosis, prognosis, and therapy. Herein we develop a distinctive label-free "off-on" configuration for photoelectrochemical (PEC) sensing platform fabrication, coupled with DNA four-way junction (4J) architecture as well as G-wire superstructure for signal amplification. In addition, ultrathin copper phosphate nanosheets (CuPi NSs) coating Au nanoparticles (Au-CuPi NSs) serve as a highly efficient photocathode substrate. To improve the sensitivity, and avoid the false positive signals, the quencher, gold nanoparticles (GNPs), is utilized to switch off the PEC signal because of the commendable surface plasmon resonance (SPR) absorption. Subsequently, ingenious DNA 4J architecture is applied to export proportional c-myc regions for target quantification. Assisted with the G-wire superstructure formation, the enhancer 5,10,15,20-tetra(4-sulfophenyl)-21H,23H-porphyrin (TSPP) is coupled on the substrate to switch on the PEC signal, thus realizing the miRNA assay with persuasive accuracy, high sensitivity, and low detection limit. In addition, we execute the miRNA detection in prostate carcinoma cell line 22Rv1, and acquire desirable quantitative capability. Remarkably, the prepared PEC sensing platform not only realizes the highly efficient miRNAs quantification, but also uncovers a marvelous horizon for sensing platform fabrication.

The 22Rv1 prostate cancer cell line carries mixed genetic ancestry: Implications for prostate cancer health disparities research using pre-clinical models.

Understanding how biological factors contribute to prostate cancer (PCa) health disparities requires mechanistic functional analysis of specific genes or pathways in pre-clinical cellular and animal models of this malignancy. The 22Rv1 human prostatic carcinoma cell line was originally derived from the parental CWR22R cell line. Although 22Rv1 has been well characterized and used in numerous mechanistic studies, no racial identifier has ever been disclosed for this cell line. In accordance with the need for racial diversity in cancer biospecimens and recent guidelines by the NIH on authentication of key biological resources, we sought to determine the ancestry of 22RV1 and authenticate previously reported racial identifications for four other PCa cell lines. We used 29 established Ancestry Informative Marker (AIM) single nucleotide polymorphisms (SNPs) to conduct DNA ancestry analysis and assign ancestral proportions to a panel of five PCa cell lines that included 22Rv1, PC3, DU145, MDA-PCa-2b, and RC-77T/E. We found that 22Rv1 carries mixed genetic ancestry. The main ancestry proportions for this cell line were 0.41 West African (AFR) and 0.42 European (EUR). In addition, we verified the previously reported racial identifications for PC3 (0.73 EUR), DU145 (0.63 EUR), MDA-PCa-2b (0.73 AFR), and RC-77T/E (0.74 AFR) cell lines. Considering the mortality disparities associated with PCa, which disproportionately affect African American men, there remains a burden on the scientific community to diversify the availability of biospecimens, including cell lines, for mechanistic studies on potential biological mediators of these disparities. This study is beneficial by identifying another PCa cell line that carries substantial AFR ancestry. This finding may also open the door to new perspectives on previously published studies using this cell line.

Co-assembled hybrids of proteins and carbon dots for intracellular protein delivery

Silver nanoparticles (AgNPs) are accumulated in the male reproductive organs for a long time and cause several adverse effects in there. Up to now, there is little of information for the cytotoxic effects in male reproductive cells. In this study, the stable AgNPs with a minimal silver ion (Ag+) dissolution below concentration inducing cytotoxicity in the cell medium were exposed to the human prostate carcinoma cell line 22Rv1. Moreover particle uptake and androgen receptor (AR) transactivation were evaluated. In cell medium, AgNPs exhibited stability in an aqueous environment and minimal Ag+ release. Transmission electron microscopy (TEM) and energy dispersive spectrometer (EDS) analysis demonstrated uptake of AgNPs into cells via endocytosis, and a quantitative Ag assay showed that uptake of AgNPs was size-dependent with the majority of Ag retained in the particle form. To evaluate if the presence of AgNPs can change androgenic potentials of dihydrotestosterone (DHT, strong human androgen), we conducted an AR transactivation assay using the transgenic prostate cell line 22Rv1-MMTV-Hyg and found that AgNPs lowered androgenic transactivation of DHT, which is due to decreased bioavailability of DHT.

Targeting nano drug delivery to cancer cells using tunable, multi-layer, silver-decorated gold nanorods.

Multifunctional nanoparticles have high potential as targeting delivery vehicles for cancer chemotherapy. In this study, silver-decorated gold nanorods (AuNR\Ag) have been successfully used to deliver specific, targeted chemotherapy against breast cancer (MCF7) and prostate carcinoma (PC3) cell lines. Doxorubicin, a commonly used chemotherapy, and anti-Epithelial cell adhesion molecule (anti-EpCAM) antibodies were covalently bonded to thiolated polyethylene glycol-coated AuNR\Ag, and the resultant system was used to deliver the drugs to cancer cells in vitro. Furthermore, these nanoparticles have a unique spectral signature by surface enhanced Raman spectroscopy (SERS), which enables reliable detection and monitoring of the distribution of these chemotherapy constructs inside cells. The development of interest in a plasmonic nano drugs system with unique spectroscopic signatures could result in a clinical approach to the precise targeting and visualization of cells and solid tumors while delivering molecules for the enhanced treatment of cancerous tumors.

Phytochemical Study of the Genus Salpichroa (Solanaceae), Chemotaxonomic Considerations, and Biological Evaluation in Prostate and Breast Cancer Cells.

Twelve Salpichroa taxa have been phytochemically analyzed. From the aerial parts of S.scandens, four known salpichrolides A, C, I, S, and an unreported withanolide named salpichrolide V (1), were isolated. In S.dependens, S.gayi, S.glandulosa subsp. glandulosa, S.glandulosa subps. weddellii, S.leucantha, S.micrantha, S.microloba, S.proboscidea, S.ramosissima, S.tristis var. tristis, and S.weberbauerii, no withanolides were found. The chemical content of ca. 85% of the Salpichroa taxa is in agreement with molecular studies, which suggest that Salpichroa and Jaborosa, a genus considered morphologically close to Salpichroa, are distant in the systematic of the Solanoideae subfamily. Moreover, the invitro cytotoxic activity of a set of natural salpichrolides and derivatives was examined against two prostate carcinoma cell lines (PC3 and LNCaP) and two human breast cancer cell lines (MCF-7 and T47D). Several compounds showed moderate activity (IC50 =64.91-29.97μm).

Expression and purification of TAT-NDRG2 recombinant protein and evaluation of its anti-proliferative effect on LNCaP cell line

N-myc downstream regulated gene2 (NDRG2) belongs to tumor suppressor protein family of NDRG. Anti-proliferative and anti-metastasis of NDRG2 overexpression has been demonstrated in a number of tumors. The aim of this study was to fuse the gene of Trans Activator of Transcription (TAT) protein transduction domain with NDRG2 gene and express and purify TAT-NDRG2 fusion protein in order to investigate the effects of TAT-NDRG2 protein on proliferation and apoptosis of LNCaP prostate carcinoma cell line. pET28a-TAT-NDRG2 and pET28a-NDRG2 plasmids were constructed and transformed into E. coli-BL21(DE3). TAT-NDRG2 and NDRG2 proteins were expressed in the bacteria, purified using affinity chromatography and verified using western blotting. The effects of TAT-NDRG2 and NDRG2 protein treatment on LNCaP cells proliferation and apoptosis were evaluated using MTT assay and AnnexinV, 7-AAD flow cytometry assay, respectively. Western blot analysis confirmed the expression and purification of TAT-NDRG2 and NDRG2 proteins. Treatment of LNCaP cells with TAT-NDRG2 protein increased cell death and induced apoptosis significantly (P < 0.05) compared to control and NDRG2 protein-treated cells. These results suggest that TAT-NDRG2 protein can be considered as a therapeutic modality for the treatment of tumors.