TP53 Inhibited AR Signaling Promote Enzalutamide Resistance in LNCaP cell

Background: To perform gene screening and search key knockout gene associated with enzalutamide resistance in LNCaP cells. Methods: Genome-scale CRISPR-Cas9 transfection were done to screen genes associated with enzalutamide resistance in LNCaP cells. Bioinformatic analysis of TP53 and AR expression correlated with prostate adenocarcinoma by using TCGA database. Knockout interfering RNA (shRNA) were synthesized in lentivirus and transfected into LNCaP cells by using Lipofectamine 3000. TP53 and AR detection was performed by using conventional molecular biology methods, such as western blot, RT-PCR and flow cytometer et al. Results: 27 high (NES<-1.7, P<0.01) and 26 low (NES>1.7, P<0.01) expressed genes were identified in enzalutamide resistance Cells. TP53 interacted with AR may play an important role in the development of enzalutamide resistance from PPI and RNA-seq data. Data from TCGA support that patient with TP53&AR low expression endured higher pathology stage, Gleason grades, more disease progress, recurrence, positive lymphnotes examination numbers and more abnormal bones scan results. LNCaP cell dealt with enzalutamide or with TP53 knockout showed inhibited cell cycle at G0~G1, increased apoptosis percentage and also inhibited cell proliferation. Conclusions: Current data support that TP53 protein inhabiting AR signaling promote enzalutamide resistance in LNCaP cell.

Investigating the Efficacy of Potential Diet-Derived Phytochemicals on LNCaP Cell Lines Under Physiological Conditions

Prostate cancer (PCa) is increasingly prevalent in Nigeria and chemoprevention has a potential role to stem the initiation, promotion and progression of prostate carcinogenesis. The in vivo efficacy of phytochemicals depends on bioavailability, as the concentration of the compounds reaching target sites are known to reduce tremendously. With lower concentrations being achieved in target organs, the outcome on key players of carcinogenesis needs to be understood. The effect of curcumin, 3,3’-Diindolylmethane and epigallocatechin-3-gallate were interrogated on LNCaP cell lines by imitating in vivo conditions. The LNCaP cell lines were firstly, treated with low doses of curcumin, DIM and EGCG respectively. Subsequently, to investigate chemopreventive potentials of selected phytochemicals, cell lines were pre-treated and subsequently stimulated with testosterone. Lastly, to investigate the therapeutic potential of selected phytochemicals, cell lines were pre-stimulated before receiving respective treatments. From results, the effect of low dose treatments on the androgen receptor (AR) was dose-dependent. AR inhibition was observed more with cell lines that received a pre-treatment than cell-lines that received a pre-stimulation. All findings indicate that the investigated phytochemicals are potential chemopreventive regimens than curative regimens, since inhibitory effects on AR were enhanced more with a pre-treatment than a pre-stimulation. Furthermore, increased bioavailability of chemopreventive regimens will enhance efficacy.

Characterization of camptothecin by analytical methods and determination of anticancer potential against prostate cancer

Background Objective of present research work is to develop and validate cost-effective analytical tool for determination of camptothecin (CPT) and determine its anticancer potential against prostate cancer LNCaP cell lines. Structural elucidation has been performed by mass spectrometry, Fourier transform infrared spectroscopy, nuclear magnetic resonance spectroscopy, and MTT assay utilized for in vitro cytotoxicity where spectrometric method was used for estimation of camptothecin. Results Mass spectra showed peak at 349.2 which matches to standard molecular weight of camptothecin. FTIR and NMR spectra conformed functional moieties and structure of isolated camptothecin which was nearly equal to values mentioned in standard structure of camptothecin. IC50 values of CPT against LNCaP cell lines was found to be 3.561 μg/ml. Lambda max of CPT was found to be at 225 nm and calibration curve found to be linear over the concentration range from 2 to 70 μg/ml of camptothecin. Developed method was found to be linear, accurate, and precise. LOD and LOQ were found to be 0.0524 μg/ml and 0.1614 μg/ml, respectively. Developed method has % relative standard deviation less than one which is reproducible hence % recovery was found to be 99.80%. Conclusions FTIR, NMR, and mass spectrometry results conforms isolated compound was camptothecin; cytotoxicity study proves it has strong potential in treatment of prostate carcinoma as competent alternative to chemotherapy in the form of herbal medicine and the developed UV method proves to be valid, sensitive, and applicable for rapid, accurate, precise, and economical determination of camptothecin.

Evaluation of Biological Activity of Different Wavelengths of Low-Level Laser Therapy on the Cancer Prostate Cell Line Compared With Cisplatin

Introduction: Cancer is one of the most important problems in the world. Low-level laser therapy (LLLT) has been emerged as a new approach, having both stimulation and inhibition effects on cellular function. The goal of this study was to analyze and compare the different concentrations of cisplatin and wavelengths of laser therapy on the LnCap cell lines. Methods: LnCap cells were cultured and treated with different concentrations of cisplatin (0.1, 0.4, 0.8, 1.2 and 2 µg/mL for 24 hours) and wavelengths of laser therapy (610, 630 and 810 nm) (0.45 J/cm2 ) separately. The viability of cells was examined by MTT assay and IC50 was also calculated. Furthermore, a combination of cisplatin IC50 (24 hours) and different wavelengths of the laser was examined. Results: The results of this study showed that 2 µg/mL of cisplatin has the most significant reduction effect on the cell viability of the LnCap cell line. Cisplatin decreased the viability of cells in a dose-dependent manner. Moreover, IC50 of cisplatin was 1.24 µg/mL. On the other hand, LLLT with wavelengths of 610, 630 and 810 nm did not show notable biological effects on cell viability. Conclusion: As known, cisplatin has the capability to reduce the viability of LnCap cell lines. However, LLLT cannot be a remarkable option for the treatment of prostate cancer. Therefore, although laser therapy showed praiseful therapeutic activity against some cancer cell lines, in this study the results indicated that defined laser wavelengths had no inhibitory effects against the prostate cancer cell line.

Circular RNA Phosphatidylinositol-4-Phosphate 5-Kinase Type 1 Alpha Affects Prostate Cancer Lymph Node Carcinoma of Prostate-Cell Multiplication and Apoptosis via Regulating MicroRNA-299-3p Expression

Prostate cancer is a common cancer in aging men. This research explores the its molecular mechanisms of the circular RNA (circRNA) circPIP5K1A and its role in the multiplication and apoptosis of prostate cancer LNCaP cells. Cancerous tissue and adjacent non-cancerous tissue samples were selected from 37 prostate cancer patients hospitalized from January 2016 to January 2020. Quantitative reverse transcription PCR (RT-qPCR) was used to amplify circPIP5K1A and detect the miR-299-3p levels. Cells from the LNCaP prostate cancer cell line were divided into the si-circPIP5K1A group, si-negative control (NC) group, miR-299-3p group, miR-NC group, si-circPIP5K1A + anti-miR-299-3p group, and si-circPIP5K1A + anti-miR-NC group. Microculture tetrazolium (MTT) assay was used to assess the cell viability; flow cytometry was used to detect and analyze cell apoptosis; western blot was used to study the protein expression; the dual-luciferase reporter assay was used to determine the targeted regulation of miR-299-3p by circPIP5K1A. CircPIP5K1A expression in prostate cancer tissue increased when miR-299-3p expression was reduced (P < 0.05) compared to those in the adjacent tissue. After inhibition of circPIP5K1A or overexpression of miR-299-3p, the activity of LNCaP cells decreased, the apoptosis rate of LNCaP cells increased, Ki-67 and Bcl-2 expressions in LNCaP decreased, and Bax expression increased (P < 0.05). Due to CircPIP5K1A regulation of miR-299-3p expression, mir-299-3p interference reversed the effect of circPIP5K1A inhibition of LNCaP-cell multiplication and apoptosis. Inhibiting the expression of circPIP5K1A may inhibit LNCaP-cell multiplication and expedite cell apoptosis by up-regulating miR-299-3p.

Differential Expression of a Panel of Ten CNTN1-Associated Genes during Prostate Cancer Progression and the Predictive Properties of the Panel towards Prostate Cancer Relapse

Contactin 1 (CNTN1) is a new oncogenic protein of prostate cancer (PC); its impact on PC remains incompletely understood. We observed CNTN1 upregulation in LNCaP cell-derived castration-resistant PCs (CRPC) and CNTN1-mediated enhancement of LNCaP cell proliferation. CNTN1 overexpression in LNCaP cells resulted in enrichment of the CREIGHTON_ENDOCRINE_THERAPY_RESISTANCE_3 gene set that facilitates endocrine resistance in breast cancer. The leading-edge (LE) genes (n = 10) of this enrichment consist of four genes with limited knowledge on PC and six genes novel to PC. These LE genes display differential expression during PC initiation, metastatic progression, and CRPC development, and they predict PC relapse following curative therapies at hazard ratio (HR) 2.72, 95% confidence interval (CI) 1.96–3.77, and p = 1.77 × 10−9 in The Cancer Genome Atlas (TCGA) PanCancer cohort (n = 492) and HR 2.72, 95% CI 1.84–4.01, and p = 4.99 × 10−7 in Memorial Sloan Kettering Cancer Center (MSKCC) cohort (n = 140). The LE gene panel classifies high-, moderate-, and low-risk of PC relapse in both cohorts. Additionally, the gene panel robustly predicts poor overall survival in clear cell renal cell carcinoma (ccRCC, p = 1.13 × 10−11), consistent with ccRCC and PC both being urogenital cancers. Collectively, we report multiple CNTN1-related genes relevant to PC and their biomarker values in predicting PC relapse.

Antiproliferative effect of rosehip tea phenolics in prostate cancer cell lines

ObjectivesRecently, phenolic compounds (quercetin, kaempferol, ellagic acid (EA), and myricetin) as natural sources have been suggested to be used for treatment and chemoprevention of prostate cancer. Since rosehip includes the above molecules in high concentration, we set out to investigate possible anti-proliferative effect of rosehip tea on the prostate cancer cell line.MethodsThe flavonol content of rosehip tea prepared at different temperatures and time intervals was determined first and then the antiproliferative effect of tea samples was established by adding tea samples to the prostate cancer cell line (VCaP and LNCaP).ResultsQuercetin was more effective in LNCaP cell than in VCaP cell (IC50 = 20 and 200 μM, respectively). The boiled fruit shredded at minute 7 showed the highest levels of quercetin, EA and kaempferol and the boiled fruit at minute 7 had the highest levels of kaempferol and EA. The tea samples were prepared in concentrations relevant to their IC50 values, added to the VCaP and LNCaP cell lines. The antiproliferative effect of rosehip tea on VCaP cells was slightly greater than that of LNCaP cells.ConclusionEach of the flavonols exhibits an antiproliferative effect. Our data clearly indicated that rosehip as a natural source of all flavonols had an antiproliferative effect on androgen-sensitive prostate cancer. Now that it is important to use natural sources in cancer, rosehip seems to be a promising natural product to be used to treat the prostate illness.