Proliferating Cell Nuclear Antigen Positivity Research Articles

Long-Term Supplementation of Canthaxanthin Does Not Inhibit Gastric Epithelial Cell Proliferation in Helicobacter mustelae-Infected Ferrets

The effect of canthaxanthin (CX), a non-provitamin A carotenoid, on gastric epithelial proliferation was studied in ferrets colonized with Helicobacter mustelae, which causes a chronic gastritis and an increased gastric epithelial cell proliferation. Seven spayed female ferrets were dosed by gavage with CX beadlets (50 mg/kg body wt, 5 d/wk) over 2 y, whereas seven control animals were given placebo beadlets. At the end of the 2-y-period, ferrets were killed, and gastric tissues were obtained from the antrum and body regions. A cell-proliferating biomarker, proliferating cell nuclear antigen (PCNA), was assayed by immunohistochemistry. The PCNA positivity was measured by a computer-based image analysis system. Serum concentrations of carotenoids, retinoids and tocopherols were analyzed by HPLC. Serum antioxidant status was measured by the oxygen radical absorbance capacity (ORAC) assay. The PCNA positivity in both antrum and body regions was not significantly different between CX-fed ferrets and controls. Serum CX concentrations were significantly higher in CX-fed ferrets vs. controls (P<0.001), whereas levels of other carotenoids, retinoids and tocopherols were not significantly different. The ORAC values were not significantly different between groups. This study does not suggest inhibitory effects of CX on gastric epithelial cell proliferation in H. mustelae infected ferrets.

Egg Shell Membrane as an Alternative Vascular Patch for Arterial Angioplasty.

Introduction: The egg shell membrane (ESM) is always considered as waste, but recent studies have shown that it has the potential to yield rapid re-endothelialization in vitro. We hypothesized that ESM and heparin-conjugated ESM (HESM) can be used as arterial patch in a rat aortic angioplasty model. Method: Sprague-Dawley rat (200 g) abdominal aortic patch angioplasty model was used. Decellularized rat thoracic aorta (TA) patch was used as the control; ESM patch was made of raw chicken egg; heparin-coated ESM (HESM) patch was made by using dopamine; anticoagulation properties were verified using platelet adhesion tests; the TA, ESM, and HESM patches were implanted to the rat aorta and harvested at day 14; and the samples were examined by immunohistochemistry and immunofluorescence. Result: The ESM patch showed a similar healing process to the TA patch; the cells could migrate and infiltrate into both patches; there was a neointima with von Willebrand factor-positive endothelial cells; the endothelial cells acquired arterial identity with Ephrin-B2- and dll-4-positive cells; there were proliferating cell nuclear antigen (PCNA)-positive cells, and PCNA and alpha smooth muscle actin dual-positive cells in the neointima in both groups. Heparin was conjugated to the patch successfully and showed a strong anticoagulation property in vitro. HESM could decrease mural thrombus formation after rat aortic patch angioplasty. Conclusion: The ESM is a natural scaffold that can be used as a vascular patch; it showed a similar healing process to decellularized TA patch; HESM showed anticoagulation property both in vitro and in vivo; and the ESM may be a promising vascular graft in the clinic.

Idebenone regulates sirt1/Nrf2/TNF-α pathway with inhibition of oxidative stress, inflammation, and apoptosis in testicular torsion/detorsion in juvenile rats.

Testicular torsion is an emergency, mainly in newborn and adolescent males, resulting in testicular ischemia. The current study aimed to evaluate the effect of Idebenone (IDE) on testicular torsion/detorsion (T/D) in juvenile rats. Thirty-two rats were randomized into: (1) the sham group: rats received sham operations with no other interventions; (2) the IDE group: rats received idebenone (100mg/kg, i. p) without T/D; (3) the T/D group: rats underwent torsion for 2h and detorsion for 4h; and (4) the IDE+ T/D group: rats received IDE 1h before T/D. Testicular malondialdehyde (MDA), total nitrite/nitrate (NOx), total antioxidant capacity (TAC), tumor necrosis factor-α (TNF-α), caspase-3, sirtuin type 1 (Sirt1), serum interleukin-1β (IL-1β), total cholesterol, and testosterone were measured. Histological changes, nuclear factor (erythroid-derived 2)-like-2 factors (Nrf2), and proliferating cell nuclear antigen (PCNA) immuno-expressions were assessed. T/D displayed an increase in MDA, NOx, TNF-α, caspase-3, IL-1β, and total cholesterol with a significant decrease in TAC, Sirt1, and testosterone and strong positive Nrf2 and negative PCNA immuno-expressions. IDE could improve all oxidative, inflammatory, and apoptotic indicators. Therefore, IDE significantly reduced testicular ischemia-reperfusion injury in the juvenile rat testicular T/D model by limiting oxidative stress, inflammation, and apoptosis via the Sirt1/Nrf2/TNF-α pathway.

Reproductive Stages and Ovarian Histophysiology in Artibeus planirostris (Chiroptera: Phyllostomidae) in the Dry and Rainy Months in a Municipality of the Mesoregion of the Mata Pernambucana, Northeastern Brazil

In this study, reproductive stages and ovarian histophysiology were investigated in adult female Artibeus planirostris in the municipality of Vitória de Santo Antão, Pernambuco, Northeastern Brazil. They were captured over one night each month (November 2016 to November 2017). Based on the monthly average precipitation, the dry (September to February) and rainy (March to August) months were grouped. Through external body morphology, females were classified as inactive, pregnant, lactating, and post-lactating. Only the inactive (n = 13) were euthanized for morphophysiological analyzes. Blood samples were obtained for estrogen measurement and, after euthanasia, the ovaries were collected and processed by routine histological techniques. In morphometry, were considered: the area of the ovary, the presence, and the area of the ovarian follicles (unilaminar and multilaminar primary, antral, and Graafian), as well as the quantification of the corpus luteum. In immunohistochemistry, PCNA (Proliferating cell nuclear antigen) was used. The results showed pregnant females only in dry months, however inactive, lactating, and post-lactating females were collected in dry and rainy months. Estrogen levels and the presence of positive PCNA cells were higher in the rainy months. The Graafian follicle occurred only in these months, whereas the corpus luteum was found in all specimens from the dry months. Although A. planirostris has a tendency to start gestation in dry months and mating from mid to the end of the rainy months, this species showed adaptability influenced by environmental variations throughout the reproductive cycle, indicating a aseasonal polyestry.

Expression of p63 and Proliferating Cell Nuclear Antigen in Oral Submucous Fibrosis.

ABSTRACTAim:Oral squamous cell carcinoma (OSCC) may be preceded by potentially malignant disorders such as oral submucous fibrosis (OSMF). p63 can detect genetic changes in OSMF and it facilitates early detection of malignant transformation, whereas proliferating cell nuclear antigen (PCNA) is a marker of proliferation and may prove to be a useful objective indicator of the biological behavior of various tumors. The aim of this study was to investigate the expression and pattern of the intensity of p63 protein and PCNA in normal oral mucosa and OSMF using immunohistochemistry (IHC) and to correlate the expression of these biomarkers.Materials and Methods:A total of 15 archival paraffin-embedded blocks obtained from our department, which were histopathologically diagnosed early OSMF (n = 4), intermediate OSMF (n = 4), and advanced OSMF (n = 2) and normal mucosa (n = 5), were taken as the standard for comparison. p63 and PCNA positivity was analyzed using Kruskall-Wallis test followed by pairwise comparison using Mann-Whitney U test. The pattern of staining and intensity was compared using Chi-Square test for which Statistical Package for the Social Sciences (SPSS, v 22.0, IBM Corporation, Armonk, New York) was used.Results:All samples showed positive staining for p63 and PCNA. A statistically significant difference (P < 0.05) was seen between the frequency of occurrence of p63 and the PCNA pattern of expression among all the groups. The intensity of staining was mild to intense in the basal layer, as there was a progression toward the severity of the disease. Almost 75.4% correlation existed between p63 and PCNA, with high correlation and marked relationship.Conclusions:The OSMF is considered a potentially malignant disorder that has the potential to get transformed into OSCC. The malignant transformation is often associated with changes at the genetic level, and these are reflected by the altered expression of proteins. Our results showed that biomarkers such as p63 and PCNA are significant in predicting the malignant transformation in OSMF, so in future they may serve as a prognostic tool in the early detection of malignancies.

Raspberry Ketones Attenuate Cyclophosphamide-Induced Pulmonary Toxicity in Mice through Inhibition of Oxidative Stress and NF-ΚB Pathway.

Cyclophosphamide (CP) was found to have a potential toxic effect on lung tissues. Raspberry ketones (RKs) are natural antioxidant chemicals isolated from red raspberries (Rubus ideaus). They are commonly used for weight loss and obesity. The current study aimed to evaluate the possible protective effects of RKs against lung toxicity induced by CP. Mice were allocated into six groups: (1) control group; (2) CP group: received a single intraperitoneal dose of CP (150 mg/kg, i.p.); and (3–6) mice were pre-treated orally with different doses of RKs (25, 50, 100, and 200 mg/kg) for 14 consecutive days, respectively, before the administration of an intraperitoneal dose of CP (150 mg/kg, i.p.). Mice were then sacrificed under anesthesia, then lungs were removed for histopathological and biochemical investigations. A single dose of CP markedly altered the levels of some oxidative stress biomarkers and resulted in the fragmentation of DNA in lung homogenates. Histological examination of CP-treated mice demonstrated diffuse alveolar damage that involved apparent hyalinization of membranes, thickening of inter alveolar septa, and proliferation of type II pneumocytes. The immunohistochemical results of CP-treated mice revealed strongly positive Bax and weakly positive proliferating cell nuclear antigen (PCNA) staining reactivity of the nuclei of the lining epithelium of the bronchioles and alveoli. CP activated the cyclooxygenase-2/nuclear factor-kappa B pathway. However, pre-treatment with RKs significantly attenuated CP-evoked alterations in the previously mentioned parameters, highlighting their antioxidant, anti-inflammatory, and anti-apoptotic potential. RKs may be suggested to be a potential candidate to ameliorate CP-induced pulmonary toxicity.

Trolox is more successful than allopurinol to reduce degenerative effects of testicular ischemia/reperfusion injury in rats

Reperfusion surgery following testicular ischemia is a reproductive health threatening status and may result with organ dysfunction in men. The high level of reactive oxygen species (ROS) and cease of blood flow to the testis are the most important reasons of this testicular injury. Until today, numerous experimental studies reported that antioxidants might be efficient to alleviate oxidative stress induced organ dysfunction. For this purpose, in this study, we have investigated the protective effects of xanthine oxidase (XO) inhibitor, allopurinol, and ROS scavenger, trolox, in a comparative perspective in testicular ischemia reperfusion injury subjected rats. Twenty-eight adult male Sprague Dawley rats were divided into four groups of seven animals in each; control, ischemia/reperfusion (I/R), allopurinol and trolox. The rats in control group did not receive any application. Animals in I/R, allopurinol and trolox groups were subjected to 2h testicular reperfusion injury following 5h ischemia. Intraperitoneally (i.p.) 1ml isotonic, 200mg/kg allopurinol and 50mg/kg trolox were administered to the animals in these groups 30min prior reperfusion. At the end of experiment, all animals were sacrificed and blood serum malondialdehyde (MDA) levels were measured. Histological sections were obtained from the testis and stained with hematoxylin and eosin (H&E), proliferating cell nuclear antigen (PCNA) and cleaved caspase-3. Apoptotic index was evaluated with TUNEL Assay. Severe morphological degenerations, increased serum MDA, cleaved caspase-3 and TUNEL Assay positivity rate, but reduced PCNA positivity rate was observed in ischemia and reperfusion group. Morphological degenerations, MDA level, apoptotic index and PCNA positive cell rate were slightly alleviated in allopurinol administered animals compared with ischemia and reperfusion group. Protection with trolox was more successful and the results of the analysis were similar to the control group. Ischemia that leading to testicular torsion is a reproductive health affecting problem and current surgical treatment methods might be insufficient to recover testis. Various types of ROS generating mechanisms in cell are limiting protective potency of allopurinol, and cocktail administration of different ROS inhibitors might be more effective. However, our results indicate that free radical scavenger trolox might be a candidate drug to alleviate degenerative effects of testicular ischemia reperfusion injury. This is the first study that demonstrates antioxidant trolox was more successful than XO inhibitor allopurinol to protect testis against ischemia and reperfusion injury in rats.

L-Cysteine Partially Protects Against Acrylamide-Induced Testicular Toxicity.

Background:Acrylamide is a widespread substance with many areas of utilization. Acrylamide also forms a part of high-temperature-processed starchy foods. To date, numerous in vivo and in vitro studies have documented that acrylamide poses toxic effects on various organ systems.Aims:To determine the potential protective effect of L-cysteine on acrylamide-induced testicular toxicity.Study Design:Animal experimentation.Methods:We randomly divided 28 rats into four groups: control (0.9% saline), L-cysteine (150 mg/kg), acrylamide (40 mg/kg), and acrylamide+L-cysteine. After a 10-day intraperitoneal injection period, we euthanized the animals, recorded their body and testis weights, collected blood samples for serum testosterone measurement, and excised the testes for histopathological and morphometric evaluation. Immunohistochemical scoring of proliferating cell nuclear antigen and bax proteins was performed.Results:Acrylamide reduced body (p<0.01) and testis weights (p<0.05), seminiferous tubule diameter (p<0.001), and proliferating cell nuclear antigen expression (p<0.05) but increased bax protein expression (p<0.01) and the percentage of seminiferous tubules containing multinucleated giant cells (p<0.001). However, no significant change was observed in serum testosterone level of the experimental groups when compared with that of controls. L-cysteine administered with acrylamide decreased multinucleated giant cell number (p<0.001) and reversed the reduced proliferating cell nuclear antigen positivity (p<0.001) but showed no effect in restoring other parameters compared with the group treated with acrylamide alone.Conclusion:Considering the dose and duration employed, the present study concluded that L-cysteine partially protects testis against acrylamide-induced toxic effects.

Positive PCNA and Ki-67 Expression in the Testis Correlates with Spermatogenesis Dysfunction in Fluoride-Treated Rats

The present study aimed to evaluate the effect of fluoride (F) on spermatogenesis in male rats. F- at 50 and 100mg/L was administered for 70days, after which the testicular and epididymis tissues were collected to observe the histopathological structure under a light microscope. The ultrastructure of the testis and sperm was also examined via transmission electron microscopy. The apoptosis of spermatogenic cells was measured through terminal deoxynucleotidyl transferase dUTP nick end labeling staining. The expression of proliferation factors, namely, proliferating cell nuclear antigen (PCNA) and Ki-67, in the testicular and epididymis tissues, were assayed through immunohistochemistry. F- at 50 and 100mg/L significantly damaged the structure of the testis and epididymis, and the testis and sperm ultrastructure exhibited various changes, including mitochondrial swelling and vacuolization, and apsilated and raised sperm membrane. F treatment significantly increased spermatogenic cell apoptosis in the testis. PCNA (P < 0.01) and Ki-67 (P < 0.01) also presented positive expression in the testis. By comparison, no significant changes occurred in the epididymis. In summary, excessive F intake results in spermatogenesis dysfunction by damaging the testicular structure and inducing spermatogenic cell apoptosis in male rats. The positive expression level of PCNA and Ki-67 was a good response to spermatogenesis dysfunction.

Effect of Boron on Thymic Cytokine Expression, Hormone Secretion, Antioxidant Functions, Cell Proliferation, and Apoptosis Potential via the Extracellular Signal-Regulated Kinases 1 and 2 Signaling Pathway.

Boron is an essential trace element in animals. Appropriate boron supplementation can promote thymus development; however, a high dose of boron can lead to adverse effects and cause toxicity. The influencing mechanism of boron on the animal body remains unclear. In this study, we examined the effect of boron on cytokine expression, thymosin and thymopoietin secretion, antioxidant function, cell proliferation and apoptosis, and extracellular signal-regulated kinases 1 and 2 (ERK1/2) pathway in the thymus of rats. We found that supplementation with 10 and 20 mg/L boron to the drinking water significantly elevated levels of interleukin 2 (IL-2), interferon γ (IFN-γ), interleukin 4 (IL-4), and thymosin α1 in the thymus of rats (p < 0.05), increased the number of positive proliferating cell nuclear antigen (PCNA+) cells and concentrations of glutathione peroxidase (GSH-Px) and phosphorylated extracellular signal-regulated kinase (p-ERK) (p < 0.05), and promoted mRNA expression of PCNA and ERK1/2 in thymocytes (p < 0.05). However, the number of caspase-3+ cells and the expression level of caspase-3 mRNA were reduced (p < 0.05). Supplementation with 40, 80, and 160 mg/L boron had no apparent effect on many of the above indicators. In contrast, supplementation with 480 and 640 mg/L boron had the opposite effect on the above indicators in rats and elevated levels of pro-inflammatory cytokines, such as interleukin 6 (IL-6), interleukin 1β (IL-1β), and tumor necrosis factor α (TNF-α) (p < 0.05). Our study showed that supplementation of various doses of boron to the drinking water had a U-shaped dose-effect relationship with thymic cytokine expression, hormone secretion, antioxidant function, cell proliferation, and apoptosis. Specifically, supplementation with 10 and 20 mg/L boron promoted thymocyte proliferation and enhanced thymic functions. However, supplementation with 480 and 640 mg/L boron inhibited thymic functions and increased the number of apoptotic thymocytes, suggesting that the effects of boron on thymic functions may be caused via the ERK1/2 signaling pathway.

Clinical significance of expression of proliferating cell nuclear antigen and E-cadherin in gastric carcinoma.

AIMto investigate the expression of proliferating cell nuclear antigen (PCNA) and E-cadherin in gastric carcinoma and to analyze their clinical significance.METHODSA total of 146 patients were selected for this study, including 38 patients with intestinal metaplasia, 42 with dysplasia, and 66 with primary gastric cancer. In addition, 40 patients with normal gastric tissues were selected as controls. The expression of PCNA and E-cadherin was detected by immunohistochemistry. Differences in PCNA and the E-cadherin labeling indexes among normal gastric mucosa, intestinal metaplasia, dysplasia, and gastric carcinoma were compared. Subjects with normal gastric tissues were assigned to a normal group, while gastric cancer patients were assigned to a gastric cancer group. The difference in PCNA and E-cadherin expression between these two groups was compared. The relationship between expression of PCNA and E-cadherin and clinicopathological features was also explored in gastric cancer patients. Furthermore, prognosis-related factors, as well as the expression of PCNA and E-cadherin, were analyzed in patients with gastric cancer to determine the 3-year survival of these patients.RESULTSThe difference in PCNA and the E-cadherin labeling indexes among normal gastric mucosa, intestinal metaplasia, dysplasia, and gastric carcinoma was statistically significant (P < 0.05). During the transition of normal gastric mucosa to gastric cancer, the PCNA labeling index gradually increased, while the E-cadherin labeling index gradually decreased (P < 0.05). The PCNA labeling index was significantly higher and the E-cadherin labeling index was significantly lower in gastric cancer than in dysplasia (P < 0.05). The expression of PCNA was significantly higher in the gastric cancer group than in the normal group, but E-cadherin was weaker (P < 0.05). There was a negative correlation between the expression of PCNA and E-cadherin in gastric carcinoma (r = -0.741, P = 0.000). PCNA expression differed significantly between gastric cancer patients with and without lymph node metastasis and between patients at different T stages. E-cadherin expression also differed significantly between gastric cancer patients with and without lymph node metastasis (P < 0.05). High T stage and positive PCNA expression were risk factors for the prognosis of patients with gastric cancer (RR > 1), while the positive expression of E-cadherin was a protective factor (RR < 1). The sensitivity, specificity, and accuracy of PCNA positivity in predicting the 3-year survival of patients with gastric cancer were 93.33%, 38.89%, and 0.64, respectively; while these values for E-cadherin negativity were 80.0%, 41.67%, and 0.59, respectively. When PCNA positivity and E-cadherin negativity were combined, the sensitivity, specificity, and accuracy were 66.67%, 66.67%, and 0.67, respectively.CONCLUSIONCombined detection of PCNA and E-cadherin can improve the accuracy of assessing the prognosis of patients with gastric cancer.

HISTOLOGICAL AND IMMUNOHISTOCHEMICAL STUDY ON THE EFFECT OF LONG TERM TREATMENT WITH AMADOL (TRAMADOL HYDROCHLORIDE) ON THE LINGUAL

Objectives: This study aimed to evaluate the effect of Amadol (Tramadol hydrochloride) on the lingual mucosa of male albino rats through histological examination of their lingual mucosa to detect any structural changes, scanning electron microscopic examination of their lingual mucosa to detect any ultrastructural changes and immunohistochemical localization of proliferating cell nuclear antigen (PCNA).Materials and methods: Thirty adult male albino rats divided into 2 group. Group I (10 animals) served as control. Group II (20 animals) received Amadol (tramadol hydrochloride) in daily oral dose of 60 mg/kg body weight using metallic curved oropharengeal tube for 4months. Results: The light microscopic examination of the tongue of the rats treated with Amadol drug showed degenerative changes that involved the surface epithelium and lamina propria of their lingual mucosa. The immunohistochemical results of the tongue of Amadol group showed weakly positive PCNA staining reactivity of the nuclei of the basal and parabasal cells of the surface epithelium. Scanning electron microscope results indicate the histological and imunohistochemical results, as showed marked degenerative changes of dorsal surfaces of the tongue.Conclusion: Tramadol prescription, after weighing its risks and benefits, should be used with limits and under supervision.

Basic Science Liver.

Basic Science Liver.

Renoprotective Effect of Lactoferrin against Chromium-Induced Acute Kidney Injury in Rats: Involvement of IL-18 and IGF-1 Inhibition.

Hexavalent chromium (CrVI) is a heavy metal widely used in more than 50 industries. Nephrotoxicity is a major adverse effect of chromium poisoning. The present study investigated the potential renoprotective effect of lactoferrin (Lf) against potassium dichromate (PDC)-induced acute kidney injury (AKI) in rats. Beside, because previous studies suggest that interlukin-18 (IL-18) and insulin-like growth factor-1 (IGF-1) play important roles in promoting kidney damage, the present work aimed to evaluate the involvement of these two cytokines in PDC model of AKI and in the potential renoprotective effect of lactoferrin. Adult male albino Wistar rats were pretreated with Lf (200mg/kg/day, p.o.) or (300mg/kg/day, p.o.); the doses that are usually used in the experiment studies, for 14 days followed by a single dose of PDC (15mg/kg, s.c.). PDC caused significant increase in serum urea, creatinine, and total protein levels. This was accompanied with decreased renal glutathione content, and increased renal malondialdehyde, IL-18, IL-4, nuclear factor kappa B (NFκB), IGF-1, and the phosphorylated form of forkhead box protein O1 (FoxO1) levels. Moreover, normal expression IFN-γ mRNA and enhanced expression of TNF-α mRNA was demonstrated in renal tissues. Histopathological investigations provoked deleterious changes in the renal tissues. Tubular epithelial hyperplasia and apoptosis were demonstrated immunohistochemically by positive proliferating cell nuclear antigen (PCNA), Bax, and Caspase-3 expression, respectively. Pretreatment of rats with Lf in both doses significantly corrected all previously mentioned PDC-induced changes with no significant difference between both doses. In conclusion, the findings of the present study demonstrated the involvement of oxidative stress, inflammatory reactions, tubular hyperplasia and apoptosis in PDC-induced AKI. It suggested a role of IL-18 through stimulation of IL-4-induced inflammatory pathway, and IGF-1 through triggering FoxO1-induced cell proliferation. Moreover, the study revealed that Lf protected the kidney against Cr-induced AKI in rats and significantly showed antioxidant, anti-inflammatory, and anti-proliferative properties with down-regulation of IL-18 and IGF-1.

Histopathological , Enzyme - Histochemical and Immunohistochemical Diagnosis of Mast Cell Tumors in the Skin of Golden Hamster

No previous records in the available literature about the cutaneous mast cell tumors in golden hamster. Two male golden hamsters with cutaneous lumps were brought to the Institute of Veterinary Pathology of Justus Leibig University of Giessen, Germany for diagnositic examination. Cutaneous lumps were taken and processed for histological, histochemical and immunohistochemical examination. Numerous oval and polygonal tumor cells with high mitotic count and basophilic metachromatic granules were observed; while the superjacent epidermis and adnexes were normal. Many unevenly proliferating cells showed positive proliferating cell nuclear antigen (PCNA) reaction and positive chymase activity. The reactions were negative to gene encoding KIT (c-Kit), Cyclooxygenase-2 (COX-2) and tryptase. These results indicated a well- differentiated grade I subcutaneous mast cell tumor. The results are discussed in relation to subcutaneous mast cell tumors in canines.

MP55-04 THERMOTHERAPY WITH MAGNETIC CATIONIC LIPOSOMES POWERFULLY SUPPRESSES PROSTATE CANCER BONE METASTASIS IN A NOVEL RAT MODEL

You have accessJournal of UrologyProstate Cancer: Basic Research III1 Apr 2015MP55-04 THERMOTHERAPY WITH MAGNETIC CATIONIC LIPOSOMES POWERFULLY SUPPRESSES PROSTATE CANCER BONE METASTASIS IN A NOVEL RAT MODEL Daichi Kobayashi, Noriyasu Kawai, Keitaro Iida, Toshiki Etani, Taku Naiki, Ryosuke Ando, Keiichi Tozawa, Tohru Mogami, and Kenjiro Kohri Daichi KobayashiDaichi Kobayashi More articles by this author , Noriyasu KawaiNoriyasu Kawai More articles by this author , Keitaro IidaKeitaro Iida More articles by this author , Toshiki EtaniToshiki Etani More articles by this author , Taku NaikiTaku Naiki More articles by this author , Ryosuke AndoRyosuke Ando More articles by this author , Keiichi TozawaKeiichi Tozawa More articles by this author , Tohru MogamiTohru Mogami More articles by this author , and Kenjiro KohriKenjiro Kohri More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2015.02.2047AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Bone metastasis is a serious problem for prostate cancer patients, and the anticancer drug docetaxel (DTX) does not effectively treat this condition. Therefore, new therapies are urgently required. At the American Urological Association annual meetings in 2007, 2008, and 2010, we reported the therapeutic effects of our magnetite nanoparticles conjugated with cationic liposomes (MCLs). Here, we examined the therapeutic potential of MCLs for bone metastasis from prostate cancer compared to conventional DTX therapy in a novel rat model that allows tumor immunity evaluation. METHODS Prostate tumor tissues were transplanted into the femurs of 4 groups of model rats: control, MCL, DTX, and MCL + DTX. Tumors were injected with MCLs, and alternating magnetic field (AMF) irradiation was performed thrice a week. We poured 200 μl MCLs (33 mg/ml) into the tumor center just before each AMF exposure from 4 directions. DTX (10 mg/kg) was administered transcaudally. Tumor proliferation and bone destruction were evaluated by examining proliferating cell nuclear antigen positivity, computed tomography images, and CD68-positive cell numbers. Tumor immunity was evaluated by assessing interleukin-2 (IL-2), interferon-gamma (IFN-γ), and heat shock protein (HSP) 70 expression and determination of the CD8-positive lymphocyte number. RESULTS We successfully established a novel femur metastasis model of prostate cancer. This model allows long-term evaluation of bones and immune function. Tumor proliferation and bone destruction were significantly lower in the MCL and MCL + DTX groups than in the control and DTX groups. MCL thermotherapy concurrently induced necrosis and apoptosis. IL-2, IFN-γ, and HSP70 expression in the MCL and MCL + DTX groups also significantly increased, and tumor immunity was enhanced through CD8-positive lymphocyte induction. CONCLUSIONS MCL thermotherapy selectively heats MCL-injected tumor tissues, inducing necrotic and apoptotic cancer cell death, tumor immunity, and suppression of bone destruction. MCL thermotherapy was clearly more effective than DTX in treating bone metastatic prostate cancer. Therefore, combined MCL thermotherapy and DTX deserves consideration as a novel treatment for this disease. © 2015 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 193Issue 4SApril 2015Page: e674 Advertisement Copyright & Permissions© 2015 by American Urological Association Education and Research, Inc.MetricsAuthor Information Daichi Kobayashi More articles by this author Noriyasu Kawai More articles by this author Keitaro Iida More articles by this author Toshiki Etani More articles by this author Taku Naiki More articles by this author Ryosuke Ando More articles by this author Keiichi Tozawa More articles by this author Tohru Mogami More articles by this author Kenjiro Kohri More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Expression of Proliferating Cell Nuclear Antigen (PCNA) in Oral Submucous Fibrosis: An Immunohistochemical Study.

Expression of Proliferating Cell Nuclear Antigen (PCNA) well-correlates with the cell division. The suprabasal expression of PCNA is considered to be a marker of dysplastic oral mucosa indicating a special proliferative cellular state in those lesions. Oral submucous fibrosis (OSMF) is considered to be a premalignant condition with potential to get transformed into oral squamous cell carcinoma (OSCC). The present study was done to assess the PCNA expression in different grades and in different layers of epithelium of OSMF and to compare the expression of PCNA in OSMF with OSCC. A group of 40 histopathologically diagnosed, formalin fixed, paraffin embedded tissue samples were included in study. The study group was further divided into 2 groups; 30 OSMF and 10 OSCC. Theses samples were collected from Department of Oral and Maxillofacial Pathology, Government Dental College and Research Institute, Bangalore. The samples were subjected to immunohistochemical method using indirect immunoenzyme LSAB method. All 40 cases showed positivity for PCNA. The percentage of positive cells was more in basal and suprabasal layer both in OSMF and OSCC, whereas the greater percentage of positive cells was seen in the superficial layer of OSCC. All OSMF cases showed positive expression in basal and suprabasal layer and only 77% of cases showed positive PCNA expression in the superficial layer with variable immunoreactivity and 23% did not show any positive staining in the superficial layer. The percentage of positive cells and the intensity of staining increased as the disease severity increased in OSMF. Increased proportion of cells expressed PCNA staining which are in proliferative layers of OSMF. This biomarker provides an insight into the biological behaviour of the condition that substantiates its potentially malignant nature and may also provide an intermediate end point in chemotherapeutic trials.

Photodynamic therapeutic role of indocyanine green in tumor-associated inflammation in skin cancer

Indocyanine green (ICG) is a promising water-soluble photosensitizer for photodynamic therapy (PDT) of tumors. It was reported to have promising phototoxic effect on different cell lines. This study aimed to evaluate the efficacy of ICG as an efficient PS agent for skin cancer induced in mice. Skin squamous cell carcinoma was induced in female CD-1 mice by 7,12-dimethylbenzanthracene and 12-O-tetradecanoyl-phorbol-13-acetate followed by an ICG/PDT treatment. The laser irradiation for PDT was adjusted to cover the whole body of the mice to make sure that the treatment protocol will be delivered to multiple tumors. The treatment of skin cancer by ICG/PDT using intravenously injected ICG initiated tumor cell death and significantly decreased cell proliferation as indicated by the reduction in proliferating cell nuclear antigen positivity. A significant reduction in the inflammatory mediators; tumor necrosis factor-α, nitric oxide and 5-lipoxygenase was reported, however the level of cyclooxygenase-2 (COX-2) was significantly elevated after ICG/PDT treatment. The proposed ICG/PDT treatment modality showed a significant anti-tumor and anti-inflammatory activity against skin cancer accompanied with COX-2 elevation.

Small Interfering RNA Targeting Nuclear Factor Kappa B to Prevent Vein Graft Stenosis in Rat Models

BackgroundIntimal hyperplasia plays an important role in vein graft stenosis. Inflammatory injury, especially nuclear factor kappaB (NF-κB) gene activation, is highly involved in stenosis progression. We examined whether neointimal hyperplasia and vein graft stenosis could be inhibited by silencing the NF-κB gene with small interference RNA (siRNA). MethodsSixty adult male Sprague-Dawley rats were randomly divided into a normal vein group, a vein graft group, a scrambled siRNA group, and an NF-κB siRNA group. We performed reverse interpositional grafting of the autologous external jugular vein to the abdominal aorta. Vein grafts were treated with liposome and gel complexes containing NF-κB siRNA or scrambled siRNA. The levels of monocyte chemoattractant protein -1, tumor necrosis factor-α, and NF-κB p65 in vessel tissues were evaluated after surgery for content of proliferating cell nuclear antigen (PCNA) and vascular wall thickness. ResultsNF-κB siRNA treated vein graft showed less neointimal formation and fewer positive PCNA cells (P < .05). In addition there were lower levels of, NF-κB p65 protein and of inflammatory mediators (P < .05) compared with the vein graft group. ConclusionOur study suggested that siRNA transfection suppressed NF-κB expression, reduced inflammatory factors, lessened neointimal proliferation, and suppressed PCNA.

Thermotherapy using magnetic cationic liposomes powerfully suppresses prostate cancer bone metastasis in a novel rat model

Bone metastasis is a serious problem for individuals with prostate cancer, and the effects of the anticancer drug docetaxel (DTX) are insufficient. We therefore examined the therapeutic potential of magnetic cationic liposomes (MCL) in a novel rat model that allows the evaluation of tumor immunity. The effects of MCL thermotherapy were compared with those of DTX as a conventional therapy for the treatment of bone metastatic prostate cancer. Prostate tumor tissues were transplanted into the femurs of model rats divided into four groups: control, MCL, DTX, and MCL + DTX. Tumors were injected with MCL, and alternating magnetic field (AMF) irradiation was performed three times a week. Tumor proliferation and bone destruction were evaluated by proliferating cell nuclear antigen positivity, computed tomography, and CD68-positive cell number, while tumor immunity was evaluated by heat shock protein (HSP) 70 expression and CD8-positive lymphocyte number. We successfully established a novel femur metastasis model of prostate cancer, and demonstrated that tumor proliferation and bone destruction in the MCL and MCL + DTX groups were significantly suppressed compared with control and DTX groups. MCL thermotherapy concurrently induced necrosis and apoptosis. The expression of HSP70 in the MCL and MCL + DTX groups was also significantly increased, and tumor immunity was enhanced through the induction of CD8-positive lymphocytes. MCL thermotherapy was clearly more effective than DTX in treating bone metastatic prostate cancer. A combination of MCL thermotherapy and DTX therefore deserves consideration as a novel treatment for this disease.