Prostate Cancer Cell Lines DU145 Research Articles

Anti-cancer activity of sirolimus loaded liposomes in prostate cancer cell lines

The aim of the study was to investigate the effect of sirolimus encapsulated liposomes for cancer therapy. The therapeutic efficacy of sirolimus loaded liposomes was assessed in two different prostate cancer cell lines LNCaP and DU145. Factors such as incubation time and lipid composition were found to play a role in the antiproliferative effect of the conventional and Stealth liposome formulations. The prepared conventional and Stealth liposomes demonstrated antiproliferative effect against LNCaP and DU145 cells while the cellular uptake also showed that the nanoparticles were internalized in the cytoplasm of the prostate cancer cells. This study provides some encouraging results for further development of sirolimus liposomes for improved therapeutic efficacy such as targeted drug delivery to cancer sites using a specific antibody.

Alsevirone-NF Reduces Serum Testosterone and Inhibits Prostate Cancer Xenograft Growth in Balb/c Nude Mice

Background: The goal of this study was to evaluate the anticancer and testosteroneinhibitory effects of 2‘-[(E) androst-5-en-17-ylidene]methyl-4‘,5‘-dihydro-1‘,3‘-oxazole-3β-oleate (Alsevirone-NF). Materials and Methods: PC-3, DU-145, LnCap and 22rv1 prostate cancer cell lines were used for MTT assay. 22rv1 subcutaneous cancer xenografts in Balb/c nude mice were used for in vivo efficacy experiments. Testosterone level was determined after repeated administration of Abiraterone 20, 100 or 200 mg/kg vs Alsevirone-NF 5, 25 or 50 mg/kg daily for 14 days. Results: Alsevirone-NF induced more significant cytotoxicity against PC3, 22rv1 and DU-145 cell lines compared to Abiraterone or Alsevirone-treated control: IC50 7.1 vs 20.6 vs 29.1 μg/ml, 7.7 vs 20.0 vs 12.7 μg/ml, 3.8 vs 43.4 vs 8.5 μg/ml, respectively. IC50 in LnCap cells was almost equal for all three studied agents, 29.2 vs 26.2 vs 30.2 μg/ml for Abiraterone, Alsevirone and Alsevirone-NF. In gonadectomized mice, significant reduction of testosterone level was observed in mice receiving Alsevirone-NF in a maximum single dose of 50 mg/kg (cumulative dose 700 mg/kg): 0.2 nmol/l vs 0.57 nmol/l in control group and 0.83 nmol/l in Abiraterone group, single dose 100 mg/kg. Statistically significant anticancer effect in vivo was obtained on day 11 after the start of treatment: Abiraterone T/C = 27% (p<0.05), Alsevirone-NF single dose 1200 mg/kg Т/С = 45% (p<0.05). Conclusion: Alsevirone-NF exhibited higher cytotoxic activity, comparable anticancer effect in 22rv1-bearing Balb/c nude mice and provided a more significant reduction of testosterone level in gonadectomized mice in direct comparison against Abiraterone.

Mechanism of the DNA Damage Repair Gene Work on Elevated α/β Value after Hypofractionated Radiotherapy for Prostate Cancer

Our previous study showed that the LQ model appeared to be inappropriate for high doses per fraction owing to α/β ratios tending to become higher when the dose per fraction increased. In order to verify this conclusion, we studied in vitro to explore the reasons for the elevated α/β ratio after single hypofractionated radiation and the effect of DNA damage repair gene work on the α/β value after different mode of fractionated radiation for prostate cancer according to the definition (α value represents irreparable lethal damage, β represents sublethal damage). We selected two prostate cancer cell lines DU145 and PC3: ① Draw the cell survival curve using colony formation assay to calculate the α/β ratios of the two cell lines, and then use BED formula to convert hypofractionated radiation (4Gy×2f and 6Gy×2f) into equivalent single hypofractionated radiation (calculated values) comparing with that on the survival curve (actual value) after different mode of hypofractionated radiation. ② Western Blot was used to detect the expression of DNA damage marker γ-H2AX and DNA repair marker RAD51 at different time points. ③ Laser confocal immunofluorescence was used to detect the foci of γ-H2AX and RAD51 after different mode of hypofractionated radiation. ① According to the cell survival curve, the α/β values of PC3 and DU145 cells were 1.75Gy and 3.78Gy, respectively. When fractionated radiation was converted into equivalent single hypofractionated radiation, the lowest calculated values of PC3 cells and DU145 cells were only 83% and 78%, respectively indicating that the ability of hypofractionated radiation was overestimated in clinic. If a larger α/β value is substituted into the BED formula, the discrepancy between the calculated and measured doses tended to become smaller. ② Compared with fractionated radiation, the results of western blot and laser confocal immunofluorescence showed that the expression of DNA damage marker γ-H2AX was higher after 30 minutes, 6 hours, and 24 hours, indicating that more DNA damage after single hypofractionated radiation. ③ The results showed that the expression of DNA repair marker RAD51 lasted for 24 hours and the DNA damage still existed after single hypofractionated radiation, indicating that there is no repairment and the damage was irreparable. And at the same time, the expression of γ-H2AX decreased after 24 hours of fractionated radiation compared with that of 6 hours, and there was no significant difference in the expression of γ-H2AX of single hypofractionated radiation between 24 hours and 6 hours, indicating that part of the repairable damage converted into irreparable damage after single hypofractionated radiation. The results of this study suggest that after single hypofractionated radiation, the irreparable damage in cells increases (that is, α value increases), and some repairable sublethal damage (β value) is converted into irreparable damage (α value). When α value increases and β value decreases, the ratio increases.

Evaluation of a New 99mTc-labeled GnRH Analogue as a Possible Imaging Agent for Prostate Cancer Detection.

Prostate cancer is a serious threat to men's health so it is necessary to develop technics for early detection of this malignancy. The purpose of this research was the evaluation of a new99mTc-labeled GnRH analogue as an imaging probe for tumor targeting of prostate cancer. 99mTc-labeled-DLys6-GnRH analogue was prepared based on HYNIC as a chelating agent and tricine/ EDDA as coligands for labeling with 99mTc. HYNIC was coupled to epsilon amino group of DLys6 through aminobutyric acid (GABA) as a linker. Radiochemical purity and stability in normal saline and serum, were determined by TLC and HPLC methods. Furthermore, calculation of protein-binding and partition coefficient constant were carried out for 99mTc labeled peptide. The cellular experiments including receptor binding specificity and affinity were studied using three prostate cancer cell lines LN-CaP, DU-145 and PC-3. Finally, the animal assessment and SPECT imaging of radiolabeled GnRH analogue were evaluated on normal mice and nude mice bearing LN-CaP tumor. The GnRH conjugate was labeled with high radiochemical purity (~97%). The radiolabeled peptide showed efficient stability in the presence of normal saline and human serum. The in vitro cellular assays on three prostate cancer cell lines indicated that the radiotracer was bound to LN-CaP cells with higher affinity compared to DU-145 and PC-3 cells. The Kd values of 99mTc- HYNIC (tricine/ EDDA)-Gaba-D-Lys6GnRH were 89.39±26.71, 93.57±30.49 and107.3±18.82 in LN-CaP, PC-3 and DU-145 cells respectively. The biodistribution studies in normal mice and LN-CaP tumor-bearing nude mice showed similar results including rapid blood clearance and low radioactivity accumulation in non-target organs. High kidney uptake proved that the main excretion route of radiopeptide was through the urinary system. The tumor uptake was 1.72±0.45 %ID/g at 1h p.i. decreasing to 0.70±0.06%ID/g at 4h p.i. for 99mTc-HYNIC-Gaba-D-Lys6GnRH. The maximum tumor/ muscle ratio was 2.30 at 1h p.i. Pre-saturation of receptor using an excess of unlabeled peptide revealed that the tumor uptake was receptor mediated. The results of the SPECT image of LN-CaP tumor were in agreement with the biodistribution data. Based on this study, we suggest LN-CaP as a favorable cell line for in vivo studies on GnRH analogues. Moreover, this report shows that 99mTc-HYNIC (tricine/EDDA)-Gaba-D-Lys6GnRH may be a suitable candidate for further evaluation of prostate cancer.

The alpha-1,2 fucosylated tubule system of DU145 prostate cancer cells is derived from a partially fragmented Golgi complex and its formation is actin-dependent

In previous work, we showed that highly proliferative cells and cancer cells, but not cells with normal growth rate, have tubules rich in alpha-1,2 fucosylated epitopes that extend radially from the nucleus to the cell periphery and form an unusual uptake system. The importance of alpha-1,2 fucosylation in forming tubules was demonstrated by proving that down-regulating the two corresponding fucosyltransferases (FUT1 and FUT2) causes tubule fragmentation. Here, we present evidence that in the prostate cancer cell line DU145, the tubules arise in actively growing cells from vesicles in the medial and trans elements of a partially fragmented Golgi complex, while in not actively growing cells the tubules become completely independent from the Golgi complex. Formation and elongation of the tubules proved to depend on the actin cytoskeleton, since the alpha-1,2 fucosylated protein(s) segregate with the cytoskeleton proteins, and not in the membrane fraction, as do the Golgi markers and other fucosylated proteins, while depolymerization of the actin filaments causes tubule fragmentation and shifting of the alpha-1,2 fucosylated proteins into the membrane fraction.

Small Molecular Inhibitors Block TRPM4 Currents in Prostate Cancer Cells, with Limited Impact on Cancer Hallmark Functions

Transient receptor potential melastatin 4 (TRPM4) is a broadly expressed Ca2+ activated monovalent cation channel that contributes to the pathophysiology of several diseases.For this study, we generated stable CRISPR/Cas9 TRPM4 knockout (K.O.) cells from the human prostate cancer cell line DU145 and analyzed the cells for changes in cancer hallmark functions. Both TRPM4-K.O. clones demonstrated lower proliferation and viability compared to the parental cells. Migration was also impaired in the TRPM4-K.O. cells. Additionally, analysis of 210 prostate cancer patient tissues demonstrates a positive association between TRPM4 protein expression and local/metastatic progression. Moreover, a decreased adhesion rate was detected in the two K.O. clones compared to DU145 cells.Next, we tested three novel TRPM4 inhibitors with whole-cell patch clamp technique for their potential to block TRPM4 currents. CBA, NBA and LBA partially inhibited TRPM4 currents in DU145 cells. However, none of these inhibitors demonstrated any TRPM4-specific effect in the cellular assays.To evaluate if the observed effect of TRPM4 K.O. on migration, viability, and cell cycle is linked to TRPM4 ion conductivity, we transfected TRPM4-K.O. cells with either TRPM4 wild-type or a dominant-negative mutant, non-permeable to Na+. Our data showed a partial rescue of the viability of cells expressing functional TRPM4, while the pore mutant was not able to rescue this phenotype. For cell cycle distribution, TRPM4 ion conductivity was not essential since TRPM4 wild-type and the pore mutant rescued the phenotype.In conclusion, TRPM4 contributes to viability, migration, cell cycle shift, and adhesion; however, blocking TRPM4 ion conductivity is insufficient to prevent its role in cancer hallmark functions in prostate cancer cells.

Selective Release of Doxorubicin from Cucurbit[8]uril Stabilized Gold Supra-Pyramid Host at pH of Small Intestine.

Gold supra-pyramid structures were obtained by the addition of acidic solution of cucurbit[8]uril (CB[8]) to an aqueous solution of citrate stabilized gold nanoparticles (AuNP). The reaction resulted in the precipitation of supra-pyramid from the solution after just 1 min of shaking. Microscopic images confirmed formation of the supra-pyramid. The stepwise structural transformation towards the supra-pyramid was examined with variable concentrations of CB[8] to AuNP solution. Anionic counter parts of these acids (Br- , NO3 - , SO4 2- and Cl- ) controlled the size of the synthesized supra-pyramids. These supra-pyramid hosts showed uptake of three anticancer drugs: oral drugs etoposide, prednisolone and intravenous drug doxorubicin. Releases of drugs from these hosts were emulated at acidic stomach pH, basic small intestinal pH and in the presence of human serum albumin (HSA). The specific release of doxorubicin was confirmed at small intestinal pH 7.4. Poor release of drugs in presence of CB[8] specific guest 1-adamantanamine confirmed the role of the supra-pyramid as the exclusive host. The release of doxorubicin from the supra-pyramid at pH 7.4 was confirmed by fluorescence microscopic imaging with prostate cancer DU-145 cell line.

Carbostyril derivatives: Synthesis of novel carbostyril-3′-carbonitrilselenophene hybrid compounds and investigation of their antiproliferative properties on prostate and breast cancer

To synthesize a new series of carbostyril-3′-carbonitrilselenophene hybrid compounds, Pechmann coumarin compounds were reacted with 2-amino-3′-carbonitrilselenophene derivatives and their anticancer activities on MCF7 and DU145 cell lines and antioxidant activities were investigated. Anticancer and antioxidant activities of the starting compounds and their corresponding new hybrid compounds were compared. It was determined that IC50 values of hybrid compounds (3e, 3f, 3g, and 3h) on the MCF7 breast cancer cell line showed the highest cytotoxic activity with 7.99, 3.64, 7.72, and 2.74 µM values, respectively. In the case of anticancer activity on the DU145 prostate cancer cell line, compounds 3f and 3h were found to have the highest cytotoxic activity with IC50 values of 4.21 and 3.90 µM, respectively. The compound 3g is also the most radical scavenging compound with an inhibition value of 88.78%.

Profiling of apoptosis-associated proteins in human prostate cancer cells in response to Montivipera bulgardaghica albizona venom by protein array

Here we studied the possible cytotoxic and apoptotic effects of Anatolian endemic mountain viper, Montivipera bulgardaghica albizona (M.b. albizona) venom on human PC-3 and DU-145 prostate cancer cell lines. M.b. albizona venom induced cytotoxicity and apoptosis of both prostate cancer cells in a concentration-dependent manner. Protein array results revealed that the levels of major components of the mitochondrion-initiated and the extrinsic death receptor-mediated pathways were significantly changed in prostate cancer cells by venom treatment. These data suggest that one or more of the purified components of the M.b. albizona venom may serve as an appropriate therapeutic agent for prostate cancer. Further bioactivity guided studies are required for the identification of bioactive compounds of M.b. albizona venom.

Cytotoxic and anti-angiogenic effects of zoledronic acid in DU-145 and PC-3 prostate cancer cell lines.

This study aims to investigate the apoptotic and anti-angiogenic effect of Zoledronic acid on hormone-refractory prostate cancer cell lines. XTT cell proliferation assay used to assess cytotoxicity. Caspase 3/7 activity and DNA fragmentation were measured to verify apoptosis. Angiogenic cytokine profiles investigated using the human angiogenesis antibody array I. Administration of Zoledronic acid on PC-3 and DU-145 prostate cancer cell lines resulted in increased cytotoxicity and apoptosis with a time- and dose-related manner. Also, the administration of Zoledronic acid significantly reduced several angiogenic cytokine productions in PC-3 and DU-145 cell lines. Zoledronic acid successfully induced apoptosis and reduced various angiogenic cytokine production in hormone-refractory prostate cancer cell lines. Novel treatment protocols may be developed in the future with chemotherapeutic combinations for the treatment of prostate cancer.

Small molecule inhibitors of the prostate cancer target KMT2D

Histone lysine N-methyltransferase 2D (KMT2D), an important methyltransferase that is involved in the methylation of lysine 4 in histone H3 (H3K4) and related to the development of prostate cancer. Hypermethylation of H3K4 is shown in prostate cancer (PCa). However, KMT2D inhibitors have not yet been developed. This article aims to design small molecule inhibitors targeting KMT2D_SET to prevent PCa cell proliferation and migration. Twenty-four inhibitors were firstly designed according to a virtual screening of computers，and shown different degrees of binding to KMT2D_SET. Compounds 1 and 16 showed high binding affinities to KMT2D, with KD values of 147 ± 32.9 μM and 176 ± 37.9 μM, respectively. In addition, they exerted strong inhibitory activity against the PCa cell lines PC-3 and DU145, with IC50 values of 1.1 ± 0.06 μM, 1.5 ± 0.06 μM and 1.8 ± 0.1 μM, 2.3 ± 0.2 μM, respectively. Furthermore, these two compounds significantly suppressed the migration of PCa cells.

Increased Tumoral Microenvironmental pH Improves Cytotoxic Effect of Pharmacologic Ascorbic Acid in Castration-Resistant Prostate Cancer Cells

BackgroundThe anticancer potential of pharmacologic ascorbic acid (AA) has been detected in a number of cancer cells. However, in vivo study suggested a strongly reduced cytotoxic activity of AA. It was known that pH could be a critical influencing factor for multiple anticancer treatments. In this study, we explored the influence of pH on the cytotoxicity of ascorbic acid. We employed castration-resistant prostate cancer (CRPC) cell lines PC3 and DU145 to observe the therapeutic effect of AA on PCa cells that were cultured with different pH in vitro. We also analyzed the influence of pH and extracellular oxidation on cytotoxicity of AA in cancer cells using reactive oxygen species (ROS) assay, cellular uptake of AA, and NADPH assay. Male BALB/c nude mice bearing prostate carcinoma xenografts (PC3 or DU145) were used to assess treatment response to AA with or without bicarbonate in vivo. The cellular uptake of AA in PCa xenografts was detected using positron emission tomography (PET). Small animal PET/CT scans were performed on mice after the administration of 6-deoxy-6-[18F] fluoro-L-ascorbic acid (18F-DFA).ResultsOur in vitro studies demonstrate that acidic pH attenuates the cytotoxic activity of pharmacologic ascorbic acid by inhibiting AA uptake in PCa cells. Additionally, we found that the cancer cell-selective toxicity of AA depends on ROS. In vivo, combination of AA and bicarbonate could provide a significant better therapeutic outcome in comparison with controls or AA single treated mice. 18F-DFA PET imaging illustrated that the treatment with NaHCO3 could significantly increase the AA uptake in tumor.ConclusionsThe alkalinity of tumor microenvironment plays an important role in anticancer efficiency of AA in CRPC. 18F-DFA PET/CT imaging could predict the therapeutic response of PCa animal model through illustration of tumoral uptake of AA. 18F-DFA might be a potential PET tracer in clinical diagnosis and treatment for CRPC.

A Profile of the Biological Activity of Functionalized Benzimidazolium Ionic Liquids

Now-a-days, the pharmaceutical industries were facing many challenges that impose the need to develop innovative and effective therapies. Hence, the investigation of effective therapies for cancer is an actual goal of every pharmaceutical industries. Recently, Ionic Liquids (ILs) have been considered as potential target in scientific community because of their unique performance in various fields. Moreover, numerous combinations made between the cation and the anion, which can be tuned the physicochemical and biological properties of ILs. Especially, these ionic liquids play a vital role for designing new drugs and producing potent bioactivity. Herein we synthesized a series of benzimidazolium based ionic liquids with different functional groups and their structure was confirmed by FT-IR, 1H NMR, 13C NMR as well as High resolution Mass spectroscopy technique antimicrobial activity of ILs were screened by five bacterial and fungal strains which was measured in terms of zone of inhibition. All the compounds were found to be effective against all bacterial species and fungi Candida albicans. Moreover, the total antioxidant capacity of ILs has been evaluated by Phosphomolybdenum method. The report indicates that kojic acid functionalized ILs exhibited significant antioxidant activity. In addition, antitumor activity of these compounds has been analyzed using human ovarian cancer cell line SK-OV-3, human prostate cancer cell line DU-145 and human leukemia cancer cell line K-562. The data obtained from antitumour studies showed that an excellent activity of ester functionalized ionic liquid confirms that can be used as an anticancer agent. However, ester functionalized and kojic acid functionalized ILs possess biologically active against the tested strains.

Synthesis of PEGylated nanographene oxide as a nanocarrier for docetaxel drugs and anticancer activity on prostate cancer cell lines.

Graphene oxide (GO) has recently been considered one of the most promising carbon derivatives in nanotechnology. It has many excellent features such as tumor targeting ability, biocompatibility and low toxicity. Therefore, we conjugated docetaxel (DTX) to GO-PEG molecule and investigate its anticancer efficacy in prostate cancer cell line (DU-145). In order to obtain GO-PEG-DTX molecules, we conjugated the DTX via bonds to PEG chains pegylated to the GO surface. We also investigated the stability of GO-PEG-DTX in different biological fluids such as cell mediums, PBS and water in vitro conditions. GO-PEG-DTX has the highest zeta potential in water. In the current research SEM, UV-Vis, and FTIR analyses and zeta potential were utilized for the characterization of nano-sized GO-PEG-DTX. Anticancer efficacy of GO-PEG-DTX were then investigated in DU-145 prostate cancer cell line using MTT metod. The prostate cancer cells were treated by different concentrations of GO-PEG-DTX, GO-PEG, GO, and DTX (1-100 µg/ml) during 24, 48 and 72 h. The spectrophotometric analyzed values at 570 nm were recorded and analysed with Graphpad Prism7. IC50 growth inhibition values was determined. The data showed that the GO-PEG-DTX had a highly effective anticancer activity on prostate cancer cell lines after 24, 48 and 72 hours compared to other molecules. GO-PEG-DTX was found statistically significant in the DU-145 cell line (***p < 0.0001, **p < 0.001, and *p < 0.01). As a result, it can be said that PEGylated GO is an excellent nanocarrier system for the high anticancer activity of DTX. Loading of anticancer drugs using this type of graphene-based nano carrier and delivery to targeted tissues may find potential application in biomedicine.

6,12-Diphenyl-3, 9-diazatetraasterane-1, 5, 7, 11-tetracarboxylate Inhibits Proliferation, Migration and Promotes Apoptosis in Ovarian Cancer Cells.

6,12-Diphenyl-3,9-diazatetraasterane-1, 5, 7, 11-tetracarboxylate (DDTC) has been synthesized by the photodimerization of 4-phenyl-1,4-dihydropyridine-3,5-dicarboxylate. The potential of theercvantitumor activity and mechanism were investigated in vitro using MTT assay in human lung cancer cell line A549, ovarian cancer cell lines SKOV3 and A2780, breast cancer cell line MCF-7, gastric cancer cell line BGC-823, colon cancer cell line HT29, prostate cancer cell line DU145, and liver cancer cell line SMMC7721. The results show that DDTC can inhibit the growth of ovarian cancer SKOV3 and A2780 cells. The best IC50 value is approximately 5.29 ± 0.38 and 4.29 ± 0.39 μM, respectively. DDTC induced the cell cycle arrest in the G2 phase by flow cytometric analysis. The migration and invasion of ovarian cancer SKOV3 and A2780 cells were inhibited by DDTC. DDTC could increase the expression protein level of E-cadherin in A2780 cells and ascend the expression protein and mRNA levels of E-cadherin in SKOV3 cells. DDTC could also decrease the protein and mRNA expression of EMT (epithelial-to-mesenchymal transition) markers of N-cadherin and Vimentin. mRNA and protein expression level of checkpoint kinase 1 (Chk1) were significantly increased and expressions of cyclin-dependent kinase (CDK1) and cell division cycle 25a (Cdc25a) were decreased in the SKOV3 and A2780 cell lines. Moreover, DDTC induced apoptosis by the cleavage and activation of caspase 3 and caspase 9.

Evaluation of the Anti-Proliferative Activity of Rare Aldohexoses against MOLT-4F and DU-145 Human Cancer Cell Line and Structure-Activity Relationship of D-Idose.

D-Allose (D-All), a C-3 epimer of D-glucose (D-Glc), is a naturally rare monosaccharide, which shows anti-proliferative activity against several human cancer cell lines. Unlike conventional anticancer drugs, D-All targets glucose metabolism and is non-toxic to normal cells. Therefore, it has attracted attention as a unique “seed” compound for anticancer agents. However, the anti-proliferative activities of the other rare aldohexoses have not been examined yet. In this study, we evaluated the anti-proliferative activity of rare aldohexoses against human leukemia MOLT-4F and human prostate cancer DU-145 cell lines. We found that D-All and D-idose (D-Ido) at 5 mM inhibited cell proliferation of MOLT-4F cells by 46 % and 60 %, respectively. On the other hand, the rare aldohexoses at 5 mM did not show specific anti-proliferative activity against DU-145 cells. To explore the structure–activity relationship of D-Ido, we evaluated the anti-proliferative activity of D-sorbose (D-Sor), 6-deoxy-D-Ido, and L-xylose (L-Xyl) against MOLT-4F cells and found that D-Sor, 6-deoxy-D-Ido, and L-Xyl showed no inhibitory activity at 5 mM, suggesting that the aldose structure and the C-6 hydroxy group of D-Ido are important for its activity. Cellular glucose uptake assay and western blotting analysis of thioredoxin-interacting protein (TXNIP) expression suggested that the anti-proliferative activity of D-Ido is induced by inhibition of glucose uptake via TXNIP-independent pathway.

Single-Cell Transcriptomics Analysis Identifies Nuclear Protein 1 as a Regulator of Docetaxel Resistance in Prostate Cancer Cells.

The majority of patients with prostate cancer treated with docetaxel develop resistance to it. To better understand the mechanism behind the acquisition of resistance, we conducted single-cell RNA-sequencing (scRNA-seq) of docetaxel-sensitive and -resistant variants of DU145 and PC3 prostate cancer cell lines. Overall, sensitive and resistant cells clustered separately. Differential gene expression analysis between resistant and sensitive cells revealed 182 differentially expressed genes common to both prostate cancer cell lines. A subset of these genes gave a gene expression profile in the resistant transcriptome-like-sensitive cells similar to the resistant cells. Exploration for functional gene pathways identified 218 common pathways between the two cell lines. Protein ubiquitination was the most differentially regulated pathway and was enriched in the resistant cells. Transcriptional regulator analysis identified 321 potential regulators across both cell lines. One of the top regulators identified was nuclear protein 1 (NUPR1). In contrast to the single-cell analysis, bulk analysis of the cells did not reveal NUPR1 as a promising candidate. Knockdown and overexpression of NUPR1 in the prostate cancer cells demonstrated that NUPR1 confers docetaxel resistance in both cell lines. Collectively, these data demonstrate the utility of scRNA-seq to identify regulators of drug resistance. Furthermore, NUPR1 was identified as a mediator of prostate cancer drug resistance, which provides the rationale to explore NUPR1 and its target genes for reversal of docetaxel resistance. IMPLICATIONS: Using single-cell sequencing of prostate cancer, we show that NUPR1 plays a role in docetaxel resistance.

Utilizing HPLC to Analyze the Presence of Anticancerous Compounds Residing from the Isolate FM1005 (Xylaria sp.) Derived from Sinularia densa

Marine organism-based natural products derived from endophytic fungi (microorganisms that reside in internal tissues of organisms and produce secondary metabolites) have been proposed as ground-breaking due to their potential to supply novel compounds for drug discovery. This investigation into possible marine-based medical applications derived from isolated compounds entailed a process inclusive of initial antiproliferative assays, the cultivation of targeted strains, and the implementation of compound separation methods for sub-fractional study and UV absorption analysis. Fractional compound analysis was conducted using a High-Performance Liquid Chromatography (HPLC) UV detector to derive probable identifications of compounds that presented antiproliferative activity. Various strains decreased the growth of A2780S (ovarian cancer), and DU-145 (prostate cancer) cell lines to the 20% and 50% viability range, respectively, and showed a degree of activity against MCF-10A (breast cancer) cell lines. Forty-eight compounds fell into the UV absorption rate range of 195-384nm, with Benzene, Rac-BINAP, and L-Histidine being commonly identified. These compound identifications can be utilized for structural analysis, novel compound discovery, and the creation of an anticancer drug that reduces the probability of cancer resurfacing in the patient. Further investigations focusing on pharmacokinetics, the biological mechanisms behind genetically inherited cancers, and the effects of specific compounds against them, such as the potential to induce apoptosis or activate inhibitory mechanisms, need to be undertaken. This research will continue to acknowledge a crucial balance point of environmental utilization, and awareness for potential future uses in the medical field.

Abstract 5344: Treatment efficacy of 212Pb-RM2 targeted alpha therapy in human prostate cancer cell lines: An in vitro investigation

Abstract Introduction: Receptor targeted alpha therapy (TAT) has garnered attention with isotopes such as Pb-212, Bi-213, and Ac-225 due to the receptor specific and potentially lethal damage caused by the in vivo alpha decay of these isotopes when linked to small peptides. We are investigating Pb-212-RM2, a BB2 receptor (BB2r) antagonist as a potential TAT agent for use in BB2r positive prostate cancer. This in vitro study evaluates Pb-212-RM2 at 3 dose levels in a panel of human prostate cancer cell lines assessing cell viability (MTT), DNA damage (53BP1), and clonogenic potential. Methods: Pb-212-RM2 was evaluated in the PC3, LNCAP, C42, C42B, DU-145, VCAP, and 22Rv1 human prostate cancer cell lines. Cells were plated the day before treatment. Pb-212-RM2 was prepared using an automated radiosynthesizer and QC was performed by HPLC. Cells were exposed to 3, 6, 13, or 25uCi/mL of Pb-212 RM2. Cell viability was evaluated at 24, 48, 72, 96, 120, and 144hrs after treatment. DNA damage was evaluated at 24hrs post treatment using the 53BP1 marker. Media in clonogenic plates was refreshed every 5-7 days until colonies of ≥50 cells were present. All assays were conducted in triplicate on different days. Results: LNCAP, C42, 22Rv1, and C42B cell lines were ≤50% viable at all dose levels by 72hrs. PC3 and DU-145 did not show a decrease to 50% viability until 96-144hrs post administration. At 144hrs post treatment, only the 25uCi/mL treated VCAP cells showed less than 50% viability (43.9%±4.1%). Observed DNA damage was highest in the PC3 and LNCAP cell lines with a range of 5.4±0.7 to 26.6±1.7 and 2.4±0.5 to 19.7±1.2 avg 53BP1 foci/cell at 3 and 25uCi/mL, respectively. Clonogenic potential was lowest in PC3 and LNCAP lines with &amp;lt;20% surviving fraction (SF) at 3uCi/mL and decreasing to &amp;lt;5% SF at 6uCi/mL. DU-145, C42B, 22Rv1, and C42 showed a SF of 71.4±11.8%, 57.1±8.8%, 55.5±2.8%, and 41.5±6.5% at 3uCi/mL. Conclusions: Targeted alpha therapy (TAT), in the form of 212Pb-RM2, was evaluated in the PC3, LNCAP, C42, C42B, DU-145, VCAP, and 22Rv1 human prostate cancer cell lines representing the spectrum of human prostate cancer in relation to androgen dependence and sensitivity. At dose levels of 3, 6, 13, or 25uCi/mL, treatment response does not appear to be related to androgen dependence or sensitivity. Cell viability decreased with increasing time and dose level in all cell lines treated with 212Pb-RM2. The VCAP cell line demonstrated the most resistance to Pb-212-TAT. DNA damage, using the 53BP1 marker, was highest in the PC3 and LNCAP cell lines. As expected, this sensitivity was also demonstrated in clonogenic response with the PC3 and LNCAP cell lines exhibiting the lowest surviving fraction. These studies provide the foundation for further exploration of the therapeutic efficacy of 212Pb-RM2 for the treatment of BB2r positive prostate cancer. This work was supported by USVA IK6BX004856, USVA IO1BX001699, and NCI RO1CA222293. Citation Format: Tammy L. Rold, Nkemakonam C. Okoye, Thomas P. Quinn, Timothy J. Hoffman. Treatment efficacy of 212Pb-RM2 targeted alpha therapy in human prostate cancer cell lines: An in vitro investigation [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5344.

QSAR, QSTR, and molecular docking studies of the anti-proliferative activity of phenylpiperazine derivatives against DU145 prostate cancer cell lines

BackgroundProstate cancer is the most common non-cutaneous cancer in males and accounts for about 4% of all cancer-related deaths in males annually. In silico methods provide faster, economical, and environmentally friendly alternatives to the traditional trial and error method of lead identification and optimization. This study, therefore, was aimed at building a robust QSAR and QSTR model to predict the anti-proliferate activity and toxicity of some phenylpiperazine compounds against the DU145 prostate cancer cell lines and normal prostate epithelial cells as well as carry out molecular docking studies between the compounds and the androgen receptor.ResultsGenetic Function Algorithm–Multilinear Regression approach was employed in building the QSAR and QSTR model. The QSAR model built had statistical parameters R2 = 0.7792, R2adj. = 0.7240, Q2cv = 0.6607, and R2ext = 0.6049 and revealed the anti-proliferate activity to be strongly dependent on the molecular descriptors: VR3_Dzp, VE3_Dzi, Kier3, RHSA, and RDF55v. The QSTR model, on the other hand, had statistical parameters R2 = 0.8652, R2adj. = 0.8315, Q2cv = 0.7788, and R2ext = 0.6344. The toxicity of the compounds was observed to be dependent on the descriptors MATS8c, MATS3s, ETA_EtaP_F, and RDF95m. The molecular descriptors in both models were poorly correlated (R < 0.4) and had variance inflation factors < 3. Molecular docking studies between the androgen receptor and compounds 25 and 32 revealed the compounds primarily formed hydrogen, halogen, and hydrophobic interactions with the receptor.ConclusionFindings from this study can be employed in in silico design of novel phenylpiperazine compounds. It can also be employed in predicting the toxicity and anti-proliferate activity of other phenylpiperazine compounds against DU145 prostate cancer cell lines.