<i>In Vitro</i>Anticancer Activity of Plant-Derived Cannabidiol on Prostate Cancer Cell Lines

Cannabinoids, the active components of Cannabis sativa Linnaeus, have received renewed interest in recent years due to their diverse pharmacologic activities such as cell growth inhibition, anti-inflammatory effects and tumor regression, but their use in chemotherapy is limited by their psychotropic activity. To date, cannabinoids have been successfully used in the treatment of nausea and vomiting, two common side effects that accompany chemotherapy in cancer patients. Most non-THC plant cannabinoids e.g. cannabidiol and cannabigerol, seem to be devoid of psychotropic properties. However, the precise pathways through which these molecules produce an antitumor effect have not yet been fully characterized. We therefore investigated the antitumor and anti-inflammatory activities of cannabidiol (CBD) in human prostate cancer cell lines LNCaP, DU145, PC3, and assessed whether there is any advantage in using cannabis extracts enriched in cannabidiol and low in THC. Results obtained in a panel of prostate cancer cell lines clearly indicate that cannabidiol is a potent inhibitor of cancer cell growth, with significantly lower potency in non-cancer cells. The mRNA expression level of cannabinoid receptors CB1 and CB2, vascular endothelial growth factor (VEGF), PSA (prostate specific antigen) are significantly higher in human prostate cell lines. Treatment with Cannabis extract containing high CBD down regulates CB1, CB2, VEGF, PSA, pro-inflammatory cytokines/chemokine IL-6/IL-8. Our overall findings support the concept that cannabidiol, which lacks psychotropic activity, may possess anti-inflammatory property and down regulates both cannabinoid receptors, PSA, VEGF, IL-6 and IL-8. High CBD cannabis extracts are cytotoxic to androgen responsive LNCaP cells and may effectively inhibit spheroid formation in cancer stem cells. This activity may contribute to its anticancer and chemosensitizing effect against prostate cancer. Cannabidiol and other non-habit forming cannabinoids could be used as novel therapeutic agents for the treatment of prostate cancer.

Induction of protective antitumor activity of tumor lysate-pulsed dendritic cells vaccine in RM-1 prostate cancer model

Objective: To investigate the antitumor activity of tumor lysate-pulsed dendritic cells vaccine in RM-1 prostate cancer mice model with the survival time of mice calculated and the tumor size measured in DC vaccine therapy. Methods: C57BL/6 mice were immunized on the dorsal flank by s.c. inoculation of Lysate-DC, ova-DC, and non-DC on day -7. On day 0, 2×106cells of RM-1 tumor cells (H-2b) were injected s.c. in C57BL/6 mice pre-treated by s.c. inoculation of modified DCs, correspondingly. DTH assay was performed with modified DCs. In partial test, for the determination of which immune cells were required for antitumor activity, mice were immunodepleted of CD4, CD8, or natural killer (NK) NK1.1 cells with the corresponding monoclonal antibodies. The survival time of nude mice loaded with tumor cells was calculated and the size of tumor measured. Results: In RM-1 mice prostate cancer model, immunized with lysate-DC, compared with ova-DC and non-DC, the pre-infection vaccine resulted in 100% clearance of primary tumors, whereas on day 0 of injection vaccine cleared 40-60% of primary tumors. On day 0, C57BL/6 mice (H-2b) were immunized with Lysate-DC, compared with ova-DC and non-DC by caudal vein injection, then on day 15, RM-1 cells were inoculated. On day 30, average diameters of tumor in different groups of modified DC were 23.7±5.4 mm, 22.1±4.9 mm, 4.3±2.6 mm, respectively. Lysate-DC, compared with ova-DC and non-DC, can greatly depressed RM-1 tumor cell growth (P<0.01). The mean survival time of C57BL/6 mice in Lysate-DC, ova-DC and non-DC groups were 15.8±2.6, 16.6±3.2, 39.0±5.6, respectively, and there was a significant difference in the mean survival time in lysate-DC group between ova-DC and non-DC group (P<0.01). DTH test showed that lysate-DC could prime T lymphocyte and elicit tumor antigen specific immune response, and over 80% mice in groups of lysate-DC showed obvious swelling in their foot pad. This response was strengthened with repeating inoculation, whereas DTH response was not seen in control group. In vivo depletion of NK cells resulted in a 40-60% reduction in growth suppression within the primary tumor, and depletion of CD4+ cells resulted in a 20% reduction in growth suppression. Conclusion: The tumor lysate-pulsed dendritic cells vaccine could elicit antitumor activity in RM-1 loaded C57BL/6 mice, and prolong the duration of RM-1 loaded C57BL/6 mice. So DC-based immunotherapy with hormone-refractory prostate carcinoma yielded protective immunity, generated efficient cellular antitumor responses, thereby providing further preclinical support for feasible immunotherapy approaches for prostate cancer.

Contribution of Anti-p63 Antibodies in the Interpretation of Benign Label Prostatic Biopsies

Introduction: Prostate cancer is the second most common cancer in men. The diagnosis is most often based on the prostate biopsies’ analysis and on histological criteria recognizable on standard coloring. In some cases, the use of immunohistochemistry is important. Objectives: This paper aims to specify the p63 phenotypic profile of lesions diagnosed benign, with minimal suspect foci, difficult to interpret, HGPIN (high grade intraepithelial neoplasia) and LGPIN (low-grade prostatic intraepithelial neoplasia) and evaluate the manual technique of p63 immunohistochemistry. Patients and Method: This was a retrospective, descriptive study of prostate biopsies recorded in the PAC service of the HALD from January 1st, 2018 to December 31st, 2018. It was completed by a manual immunohistochemical study of the blocks enrolled from November 19th to December 4th, 2020 in the PAC department of the HPD. The studied parameters were: registry number, age, clinical stage, prostate volume, PSA level, microscopic appearance and p63 immunohistochemical profile. Results: Our study included 60 prostate biopsies. The ages of our patients varied from 45 to 77 years, with an average of 64.2 years and a standard deviation of 6.2. The majority of patients were at clinical stage cT2b (33%) with a prostate volume varying between 33.15 and 169.4 cc. The minimum value of PSA in our series is 5 ng/ml, the maximum being 100 ng/ml with an average level of 24.1 ng/ml and a standard deviation of 21.2. Our series included 50 adenomyomatous hyperplasias, 7 adenomyomatous hyperplasias associated with chronic prostatitis, 2 HGPIN and 1 LGPIN. After re-reading we found 5 discordant cases, which corresponded to minimal suspect foci (kappa = 0.5098). The p63 marking was informative in 53 cases, i.e. 88%, and non-informative in 7 cases, i.e. 12%. Among the uninformative markings, 2 were due to lack of tissue adhesion to the slides. Among the informative markings, 11 were negative. p63 immunohistochemistry was useful in all suspected foci and detected 6 other minimal foci of adenocarcinoma. Conclusion: The immunostaining with the anti-p63 antibody in the prostate cancer diagnosis is of considerable benefit. It made it possible to correct 11.3% of benign diagnosis in minimal malignant focus in our context. Despite the difficulties associated with the manual technique, it is possible to have an informative rate, similar to the automatic technique.

The Combination of Artesunate and Paclitaxel in 1:1 Ratio Induces Apoptosis and Morphology Change on Human Prostate Cancer Cell Lines

The combination of Artesunate (ART) and Paclitaxel (PTX) in two human prostate cancer (PCa) cell lines (PC-3 and LNCaP) was evaluated to investigate the effects on proliferation, apoptosis and morphological changes. The half maximal inhibitory concentration (IC50) values that were observed by ART and PTX on both LNCaP and PC-3 cell lines at 72-and 120-hour exposure were used to assess these effects. Early and late apoptosis was detected in Annexin V-FITC/PI assay revealed a shift in population of cells towards early and mid-apoptosis with ART + PTX than with ART and PTX individually. More effects were observed on LNCaP cell lines at both 72-hour and 120-hour exposure. The results for the Caspase 3/7 activity assay showed shift of viable population in all induced samples compared to control. Morphological changes occurred in both cell lines;this was validated in qualitative assessment when examined under the inverted microscope. These findings indicated that ART + PTX suppressed PCa cell proliferation in a dose- and time-dependent manner.

Contemporary approach to active surveillance for favorable risk prostate cancer

The approach to favorable risk prostate cancer known as“active surveillance”was first described explicitly in 2002.This was a report of 250 patients managed with a strategy of expectant management,with serial prostate-specific antigen and periodic biopsy,and radical intervention advised for patients who were re-classified as higher risk.This was initiated as a prospective clinical trial,complete with informed consent,beginning in 2007.Thus,there are now 20 years of experience with this approach,which has become widely adopted around the world.In this chapter,we will summarize the biological basis for active surveillance,review the experience to date of the Toronto and Hopkins groups which have reported 15-year outcomes,describe the current approach to active surveillance in patients with Gleason score 3þ3 or selected patients with Gleason score 3þ4 with a low percentage of Gleason pattern 4 who may also be candidates,enhanced by the use of magnetic resonance imaging,and forecast future directions.

Role of serial multiparametric magnetic resonance imaging in prostate cancer active surveillance

AIM:To examine whether addition of 3T multiparametric magnetic resonance imaging(mp MRI)to an active surveillance protocol could detect aggressive or progressive prostate cancer.METHODS:Twenty-three patients with low risk disease were enrolled on this active surveillance study,all of which had Gleason score 6 or less disease.All patients had clinical assessments,including digital rectal examination and prostate specific antigen(PSA)testing,every 6 mo with annual 3T mp MRI scans with gadolinium contrast and minimum sextant prostate biopsies.The MRI images were anonymized of patient identifiers and clinical information and each scan underwentradiological review without the other results known.Descriptive statistics for demographics and follow-up as well as the sensitivity and specificity of mp MRI to identify prostate cancer and progressive disease were calculated.RESULTS:During follow-up(median 24.8 mo)11 of 23 patients with low-risk prostate cancer had disease progression and were taken off study to receive definitive treatment.Disease progression was identified through upstaging of Gleason score on subsequent biopsies for all 11 patients with only 2 patients also having a PSA doubling time of less than 2 years.All 23 patients had biopsy confirmed prostate cancer but only 10 had a positive index of suspicion on mp MRI scans at baseline(43.5% sensitivity).Aggressive disease prediction from baseline mpM RI scans had satisfactory specificity(81.8%)but low sensitivity(58.3%).Twentytwo patients had serial mp MRI scans and evidence of disease progression was seen for 3 patients all of whom had upstaging of Gleason score on biopsy(30% specificity and 100% sensitivity).CONCLUSION:Addition of mp MRI imaging in active surveillance decision making may help in identifying aggressive disease amongst men with indolent prostate cancer earlier than traditional methods.

Outcomes of combination MRI-targeted and transperineal template biopsy in restaging low-risk prostate cancer for active surveillance

Objective:Active surveillance(AS)offers a strategy to reduce overtreatment and now is a widely accepted treatment option for low-risk prostate cancer.An ideal tool for risk-stratification would detect aggressive cancers and exclude such men from taking up AS in the first place.We evaluate if a combination of transperineal template biopsy with magnetic resonance imaging(MRI)-targeted biopsy identifies significant prostate cancer amongst men initially diagnosed with low-risk prostate cancer.Methods:This prospective,single-blinded study included men with low-risk prostate cancer(D’Amico’s Criteria)diagnosed on conventional transrectal ultrasound-guided biopsy.Patients first underwent multiparametric MRI of the prostate
6 weeks after initial biopsy.Each suspicious lesion is mapped and assigned a Prostate Imaging Reporting and Data System(PIRADS)score.Template biopsy is first performed with the surgeon blinded to MRI findings followed by MRI-targeted biopsy using a robotic transperineal biopsy platform.Results:The age of the 19 men included is 65.4±4.9 years(mean±SD).Prostate specific antigen(PSA)at diagnosis and at the time of transperineal biopsy were comparable(7.3±1.7 ng/mL and 7.0±1.8 ng/mL,p Z 0.67),so were prostate volumes(34.2±8.9 mL and 32.1±13.4 mL,p Z 0.28).MRI-targeted biopsy had a higher percentage of cancer detection per core compared to template biopsy(11.7%vs.6.5%,p Z 0.02),this was more than 3 times superior for Gleason 7 disease(5.9%vs.1.6%,p<0.01).Four of 18(22.2%)patients with MRI lesions had significant disease with MRI-targeted biopsy alone.Three of 19 patients(15.8%)had significant disease with template biopsy alone.In combination,both techniques upclassified five patients(26.3%),all of whom underwent radical prostatectomy.Whole mount histology confirmed tumour location and grade.All six patients with PIRADS 5 lesions had cancer detected(66.6%significant disease).Conclusion:A combination of MRI-targeted and template biopsy may optimally risk-classify“low-risk”patients diagnosed on initial conventional transrectal ultrasonography(TRUS)prostate biopsy.

Expending the Paradigm: Active Surveillance for Intermediate Risk Prostate Cancer

Prostate cancer is the leading male cancer worldwide. There remains a controversy as to which patients have indolent disease and which patients present an aggressive disease needing treatment with intent to cure. Because of quality of life impairment associated with treatment by radiation or surgery, active surveillance (AS) is a valid management option to avoid or differ aggressive treatment. Traditionally, AS was reserved for men with low risk prostate cancer, however intermediate risk patients are more and more found in AS cohorts. The aim of this review is to describe the place of AS in intermediate risk patients and the perspectives offered by such a treatment modality.

Immune mediators in the tumor microenvironment of prostate cancer

Prostate cancer tissue is composed of both cancer cells and host cells.The milieu of host components that compose the tumor is termed the tumor microenvironment(TME).Host cells can be those derived from the tissue in which the tumor originates(e.g.,fibroblasts and endothelial cells)or those recruited,through chemotactic or other factors,to the tumor(e.g.,circulating immune cells).Some immune cells are key players in the TME and represent a large proportion of non-tumor cells found within the tumor.Immune cells can have both anti-tumor and pro-tumor activity.In addition,crosstalk between prostate cancer cells and immune cells affects immune cell functions.In this review,we focus on immune cells and cytokines that contribute to tumor progression.We discuss T-regulatory and T helper17 cells and macrophages as key modulators in prostate cancer progression.In addition,we discuss the roles of interleukin-6 and receptor activator of nuclear factor kappa-B ligand in modulating prostate cancer progression.This review highlights the concept that immune cells and cytokines offer a potentially promising target for prostate cancer therapy.

Content determination of benzyl glucosinolate and anti—cancer activity of its hydrolysis product in Carica papaya L.

Objective:To determine the content of benzyl glueosinolate（BG） in the pulp and the seed and investigate the anti-cuncer activity of its hydrolysis product in Curica papaya L.Methods: Determination of BG was performed on an Hypersil BDS C18 column at the wavelength of 214 nm with 0.1%trifluoroacelic acid（TFA） aqueous solution（A） and 0.1%TFA acelonilrile（B） as the mobile phase.In vitro activity test was adopted with cidtured human lung cancer H69 cell in vitro to investigate the inhibition rate of cell proliferation of benzyl isothiocyanale（BITC） againsl H69 cell.Results:The pulp has more BG before the maturation of papaya and it nearly disappeared after papaya matured,while the seed contains BG at every stage.Activity test demonstrated that the a higher concentration of BITC would have betler inhibition rate of cell proliferation on 1169 cell,and the IC50 was 6.5μmol/L.Conclusions:BG also can be produced in the pulp of papaya and it will be stored in the seed after the fruit has been matured.The hydrolysis product of BG has certain cancer-prevention anti-cancer activities for human.

Use of androgen deprivation therapy in prostate cancer： indications and prevalence

Androgens play a prominent role in the development, maintenance and progression of prostate cancer. The introduction of androgen deprivation therapies into the treatment paradigm for prostate cancer patients has resulted in a wide variety of benefits ranging from a survival advantage for those with clinically localized or locally advanced disease, to improvements in symptom control for patients with advanced disease. Controversies remain, however, surrounding the optimal timing, duration and schedule of these hormonal approaches. Newer hormonal manipulations such as abiraterone acetate have also been investigated and will broaden treatment options for men with prostate cancer, This review highlights the various androgen-directed treatment options available to men with prostate cancer, their specific indications and the evidence supporting each approach, as well as patterns of use of hormonal therapies.

Therapeutic targeting of the androgen receptor(AR)and AR variants in prostate cancer

Prostate cancer(PCa)accounted for over 300000 deaths world-wide in 2018.Most of the PCa deaths occurred due to the aggressive castration-resistant PCa(CRPC).Since the androgen receptor(AR)and its ligands contribute to the continued growth of androgendependent PCa(ADPCa)and CRPC,AR has become a well-characterized and pivotal therapeutic-target.Although AR signaling was identified as therapeutic-target in PCa over five-decades ago,there remains several practical issues such as lack of antagonist-bound AR crystal structure,stabilization of the AR in the presence of agonists due to N-terminus and C-terminus interaction,unfavorable large-molecule accommodation of the ligand-binding domain(LBD),and generation of AR splice variants that lack the LBD that impede the discovery of highly potent fail-safe drugs.This review summarizes the AR-signaling pathway targeted therapeutics currently used in PCa and the approaches that could be used in future ARtargeted drug development of potent next-generation molecules.The review also outlines the discovery of molecules that bind to domains other than the LBD and those that inhibit both the full length and splice variant of ARs.

Single Positive Core Prostate Cancer Has Less Aggressive Pathologic Features than Multiple Positive Core Prostate Cancer but Should It Still Be Considered an Indolent Tumor?

Introduction: A single positive core (SPC) in a prostate biopsy is usually associated with indolent prostate cancer (PCa), and is considered one active surveillance criteria. To determine if a SPC should qualify a patient for surveillance, we compared the pathological findings for SPC and multiple positive core in a matched population who underwent radical prostatectomy (RP). Material and Methods: We evaluated 373 SPC patients who underwent RP (Group 1) and 375 consecutive cases with multiple positive core (Group 2) who were matched according to age, prostate weight, PSA level and clinical stage. In addition to preoperative data and epidemiological characteristics, we compared the rates of positive surgical margins (PSMs), extraprostatic extension (EPE) and seminal vesicle invasion (SVI) according to the Gleason scores (GSs) of the biopsies. Results: Both groups were similar according age, PSA level, prostate weight and clinical stage. Group 1 had a lower PSM rate (20.9% vs 37.6%, p < 0.001), less EPE (10% vs 26%, p < 0.001) and SVI (6% vs 13.3%, p = 0.006). The PSM, EPE and SVI rates increased with increasing GS in both groups. In Group 1, a patient with a GS ≥ 8 was 3.5 times more likely to have a PSM than a patient with a GS ≤ 6 (p = 0.03);with no difference in Group 2 (p = 0.162). There were no correlations between the EPE and SVI rates and the GS in Group 1 (p = 0.273 and p = 0.95, respectively). However, in Group 2, we observed a higher rate of EPE among GS 7 than among GS ≤ 6, OR = 3.1 (p < 0.001). We also observed a higher rate of SVI among GS 7 than among GS ≤ 6 in Group 2, OR = 3.1 (p = 0.004). Conclusion: SPC PCa have reduced rates of PSM, EPE and SVI relative to multiple positive core. However, these pathologic findings were observed in 6.0% - 20.9% of SPC, especially in undifferentiated tumors. These results led us to conclude that active treatment instead of active surveillance should be considered and must be evaluated individually for SPC patients, especially those with higher GSs.

Pathologic and Prognostic Outcomes of Very Low- and Low-Risk Prostate Cancer According to the National Comprehensive Cancer Network Guidelines in Japanese Patients with Radical Prostatectomy

Background: The purpose of this study was to validate the treatment strategy for a cohort of Japanese patients with very low-risk (VLR) and low-risk (LR) prostate cancer according to the National Comprehensive Cancer Network (NCCN) guidelines. Methods: We studied 751 patients with T1- 3N0M0 prostate cancer treated with radical prostatectomy at our institution between 2000 and 2012. Patients with neoadjuvant treatments were excluded. We retrospectively reviewed the clinical and pathological outcomes for patients with VLR or LR prostate cancers that were classified by NCCN guidelines. Results: We identified 45 patients with VLR and 137 with LR prostate cancer. Non-biochemical recurrence rate at 5-year for 45 patients with VLR was 86.9% and 81.2% for 137 patients with LR (p = 0.56). However, none of the 19 patients >65 years old with VLR progressed, while 19% of 26 patients ≤65 years old with VLR cancer, 14% of patients >65 years old with LR cancer, and 17% of patients ≤65 years old with LR cancer progressed during the follow-up period (p = 0.04, p = 0.04 and p = 0.05, respectively). In analyses of prostatectomy specimens, both VLR and LR had similarly favorable outcomes, but patients >65 years old with VLR had the smallest tumors, with a mean of 5 mm in diameter. Conclusions: Our results support the treatment strategy of the NCCN that patients with VLR cancer and age >65 years old are good candidates for active surveillance, and that other treatment options—including active surveillance and aggressive treatments—can be applied to the remaining patients with VLR or LR cancers.

Prostate Cancer and Low Back Ache—Evidenced Role of Physiotherapy

Low back pain remains a most common clinical entity among musculoskeletal disorders. Pain reducing modalities, Manual therapy various specific techniques were widely used physiotherapeutic means as part of treatment for subjects with low back pain. An emerging trend with Independent physiotherapy practice, knowing red flags, conditions requiring investigations and experts treatment were to be recognized and adhered for maximizing patients care and benefits. Prostate cancer among men above 50 years were more found to be linked with Low back pain. This original research presentation where a subject having chronic low back pain found to have prostate cancer were analyzed and discussed with scientific evidence on clinical manifestations, investigations and medical management. Underlines the importance of recognizing, directing and getting treated of the root cause of subjects suffering with Low back pain due to prostate cancer and not just keep treating the symptoms alone were major purpose of this study.

Men with prostate cancer and the accessibility to information—a literature review

Objective: To explore possible consequences of short stays in hospitals, as these short contacts reduce the patients’ time for information and support. Method: A literature survey was carried out to get an insight in possible consequences by summarizing the state of knowledge on how men with prostate cancer undergoing prostatec-tomy surgery experience their contacts with the healthcare professionals. Results: A consequence is that often men with prostate cancer, treated with prostatectomy surgery, do not receive the individualized support, infor-mation, and dialogue they need, which leads to feelings of uncertainty, insecurity, and loss of control. The men use the Internet in their search for information and support, which makes them able to stay in control and be active, responsible partners in their own course of treatment. Conclusion: For men to feel secure and certain the accessibility of the healthcare professionals and the healthcare professionals’ ability to individualize information and support are important aspects. Practice Implications: It is relevant to provide male cancer patients with tools that can underpin their contact to the healthcare professionals. Utilizing Web 2.0 technologies, Internet based tools can support exchange-ability, towards dialogue-based contacts, between men with prostate cancer and healthcare professionals.

The current role of prostate multiparametric magnetic resonance imaging

Prostate multi-parametric magnetic resonance imaging(mpMRI)has shown excellent sensitivity for Gleason
7 cancers,especially when their volume is
0.5 mL.As a result,performing an mpMRI before prostate biopsy could improve the detection of clinically significant prostate cancer(csPCa)by adding targeted biopsies to systematic biopsies.Currently,there is a consensus that targeted biopsies improve the detection of csPCa in the repeat biopsy setting and at confirmatory biopsy in patients considering active surveillance.Several prospective multicentric controlled trials recently showed that targeted biopsy also improved csPCa detection in biopsy-naǐve patients.The role of mpMRI and targeted biopsy during the follow-up of active surveillance remains unclear.Whether systematic biopsy could be omitted in case of negative mpMRI is also a matter of controversy.mpMRI did show excellent negative predictive values(NPV)in the literature,however,since NPV depends on the prevalence of the disease,negative mpMRI findings should be interpreted in the light of a priori risk for csPCa of the patient.Nomograms combining mpMRI findings and classical risk predictors(age,prostatespecific antigen density,digital rectal examination,etc.)will probably be developed in the future to decide whether a prostate biopsy should be obtained.mpMRI has a good specificity for detecting T3 stage cancers,but its sensitivity is low.It should therefore not be used routinely for staging purposes in low-risk patients.Nomograms combining mpMRI findings and other clinical and biochemical data will also probably be used in the future to better assess the risk of T3 stage disease.

Anti-cancer effect of ethylacetate fraction from Orostachys japonicus on HT-29 human colon cancer cells by induction of apoptosis through caspase-dependent signaling pathway

Objective: To investigate the anti-colon cancer effects of ethylacetate fraction from Orostachys japonicus(0. japonicus) on HT-29 cancer cells. Methods: The viability of HT-29 cells was assayed by the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2 H-tetrazolium(MTS) method. Apoptosis induction and cell cycle inhibition were confirmed by fluorescein isothiocyanate and propidium iodide staining using flow cytometry.Morphological changes in the nucleus were observed, using a fluorescence microscope with4',6-diamidino-2-phenylindole(DAPI) nuclear staining. The expression levels of the upstream and downstream proteins involved in the anti-cancer mechanism were confirmed by Western blotting. Results: After treating HT-29 cells with different concentrations of ethylacetate fraction from O. japonicus, the viability of cells decreased in a concentration-dependent manner,while apoptosis induction and apoptotic body formation increased. Cell cycle analysis showed that the arrest occurred at the sub-G_1 and S phase. Among the upstream and downstream proteins involved in anti-cancer activity, the level of B cell lymphoma-2 decreased, and the bcl-2-associated x protein increased. The level of pro-caspase-3, pro-caspase-8, and pro-caspase-9 decreased, while the level of cleaved-caspase-3, cleaved-caspase-8, and cleaved-caspase-9 increased. Moreover, the phosphorylation, that is, activation of extracellular signal regulated kinase 1/2, Jun-N-terminal kinase, and p38 increased. Conclusions: Combining the above results, it is thought that the survival of HT-29 cells is suppressed by ethylacetate fraction from0. japonicus through mitochondrial regulation-induced caspase cascade activation, induction of apoptosis and cell cycle arrest.

Prevalence of Isolated “Pre-Malignant” Lesions on Prostate Biopsy in a Racially Diverse Community Screened Cohort

Objective: We investigated rates of prostate cancer (PCa), high-grade prostatic intraepithelial neoplaisa (HGPIN) and atypical small acinar proliferation (ASAP) in a multiethnic cohort. Methods: We evaluated prostate biopsy outcomes in men enrolled in the San Antonio Center of Biomarkers of Risk for prostate cancer (SABOR) prospective, observational study. PCa-free men underwent annual PSA testing over nearly 14 years with biopsies based on community standards. We investigated biopsy outcomes with a special interest in rates of cancer, HGPIN, and ASAP. Results: We identified 975 prostate biopsies in 801 subjects from 3/1/2001 to 1/9/2014. PCa, HGPIN, or ASAP was encountered in 28.8% (281/975), 10.1% (98/975), and 5.2% (51/975) of prostate biopsy specimens, respectively. The most significant risk factor for a PCa diagnosis was African American race (OR 5.0, 95% CI: 2.2 - 11.4, p < 0.001). HGPIN and ASAP occurred more commonly in association with PCa (both p < 0.001). We identified 57% (24/42) of men diagnosed with a “pre-malignant” lesion on prostate biopsy and had a subsequent biopsy. Of those only 8% (2/24) were diagnosed with prostate cancer (both Gleason 3 + 3) within 1 year of the initial biopsy. Conclusion: We note a 5-fold increased risk of PCa for African American men. The incidence of HGPIN and ASAP are consistent with previously reported incidence. If diagnosed in isolation, repeat biopsy within one year could be delayed or eliminated as it may not change prostate cancer outcomes.

Sulfuric Acid Catalyzed Preparation of Alkyl and Alkenyl Camptothecin Ester Derivatives and Antitumor Activity against Human Xenografts Grown in Nude Mice

Camptothecin-20-propinate (CZ48) and other camptothecin ester derivatives were prepared by the esterification reac-tions of camptothecin or 9-nitrocamptothecin with the corresponding acylating agents such as organic acid anhydride or chloride with concentrate sulfuric acid as the catalyst. The sulfuric acid-catalyzed reactions gave high yields of camptothecin ester products.Among the 11 compounds prepared by this method, camptothecin-20-O-propionate, camptothecin-20-O-crotonate, and 9-nitrocamptothecin-20-O-propionate showed good anticancer activity against various types of human tumors grown as xenografts in nude mice. The methodology developed for the preparation of camptothecin esters in this article can be applied to a wide scope of other ester derivatives.