ABSTRACT
@#[摘 要] 目的:本研究旨在探讨转录因子HOXC13在喉鳞状细胞癌(LSCC)中的表达、功能及可能的调控机制。方法:常规培养LSCC细胞,将其分为sh-NC组、sh-HOXC13组、pcDNA3.1-NC组、pcDNA3.1-HOXC13组、pcDNA3.1-PCNA组和sh-HOXC13+pcDNA3.1-PCNA组,用转染试剂将相应核酸和质粒转染各组细胞。用数据库数据分析HOXC13 mRNA在LSCC组织中的表达;收集2019年1月至2022年12月在联勤保障部队第九八〇医院耳鼻咽喉头颈外科手术切除的62对LSCC组织及配对的癌旁组织,免疫组织化学法检测中国人LSCC组织中HOXC13蛋白的表达,qPCR检测中国人LSCC组织、癌旁组织以及各组细胞中HOXC13和PCNA mRNA的表达,MTS法检测各组AMC-HN-8细胞的增殖能力,平板克隆实验检测各组AMC-HN-8细胞的克隆形成能力,Transwell小室实验检测各组AMC-HN-8细胞的迁移和侵袭能力,双萤光素酶报告基因实验和染色质免疫沉淀技术(ChIP)验证HOXC13与PCNA之间的结合关系。结果:HOXC13和PCNA在LSCC组织和细胞中均呈高表达(P<0.05或P<0.01)且两者的表达水平呈正相关(P<0.01),HOXC13的表达水平与TNM分期有明显关联(P<0.01)。敲减HOXC13可明显抑制AMC-HN-8细胞的增殖、迁移和侵袭能力(均P<0.01),过表达HOXC13则促进TU686细胞的增殖、迁移和侵袭能力(均P<0.01)。HOXC13可与PCNA启动子区结合并调控其转录。敲低PCNA可部分逆转HOXC13对AMC-HN-8细胞的恶性生物学行为的促进作用(均P<0.01)。结论:HOXC13通过上调PCNA促进LSCC细胞的恶性生物学行为,HOXC13是LSSC临床诊断和治疗的潜在靶点。
ABSTRACT
【Objective】 To investigate the role of RRM2 in prostate cancer and the mechanism. 【Methods】 The data of prostate cancer expression profile were downloaded from The Cancer Genome Atlas (TCGA). The correlation between RRM2 expression and clinicopathological features and prognosis of prostate cancer was analyzed. The protein expressions of RRM2 in 55 cases of prostate cancer and 38 benign tissues were determined with immunohistochemistry (IHC). The effects of RRM2 on the biological process of prostate cancer were assessed with bioinformatic analysis. The biological process of RRM2 affecting the progression of prostate cancer was verified with Western blot and flow cytometry. 【Results】 RRM2 was highly expressed in prostate cancer, and the expression was positively correlated with the clinical stage, pathological grade and metastasis of prostate cancer (P<0.05). Higher RRM2 expression predicted poorer survival. RRM2 co-expression positively correlated genes were involved in cell cycle pathways, pyrimidine nucleotide metabolism, and biological processes such as RNA transport. Cell cycle pathways were significantly enriched. RRM2 was highly correlated with CDK1 and PCNA molecules. RRM2 knockdown reduced the protein expressions of CDK1 and PCNA in DU145 and LNCap cell lines, which were arrested in the G2/M phase. 【Conclusion】 RRM2 promotes tumor progression by interfering with G2/M cycle of prostate cancer cells.
ABSTRACT
The extracts of Platycladus orientalis (L.) Franco leaves have shown promising anti-cancer, anti-oxidant and anti-inflammatory potency with the traditional knowledge of healing HPV associated warts. The purpose of this research is to assess the synergistic activity of sorafenib and Platycladus orientalis (L) leaf extraction on cervical cancer cells. The cytotoxicity efficiency of different concentrations of Sorafenib and ethanol extract of Platycladus orientalis (L.) leaves were tested on HeLa cells by MTT and Trypan blue assays. The synergistic effect of the IC50 concentrations of Sorafenib and Platycladus orientalis (L.) on HeLa cell by MTT assay, and mRNA expression levels of tumor suppressor tazarotene-induced gene 3 (TIG3), proliferating cell nuclear antigen (PCNA) gene and apoptosis modulator (Bcl-2) gene by RT-PCR were evaluated with individual treatments. Combination treatment showed a relatively more expression of TIG3 and less expression of Bcl-2 and PCNA was observed. Growth factor-induced MAPKP activation was arrested by compound combination treatment, which and suppression of proliferation-induced apoptosis of cervical cancer cells. Based on the our results, the combination of sorafenib and crude leaf extract from Platycladus orientalis (L.) can effectively suppress cervical cancer cell growth, thereby providing an interesting rationale for further clinical trials and in-vivo studies.
Subject(s)
Uterine Cervical Neoplasms , SorafenibABSTRACT
SUMMARY: Acrylamide (AA) is a widely used chemical and an important monomer in various industrial and laboratory processes. In addition, AA is formed during processing of starchy food at high temperature. The aim of our study was to examine effects of subchronic AA treatment on adult rat liver using histological, stereological and biochemical methods. Adult male Wistar rats were treated with AA at doses of 25 mg/kg b.w. and 50 mg/kg b.w. for three weeks. Stereological analysis showed decrease of volume density of hepatocyte cytoplasm, and increase of volume density of hepatocyte nuclei and nucleocytoplasmic ratio in AA50mg group. Immunohistochemical analysis of the liver sections showed that treatment with AA50mg increase the percentage of PCNA positive cells, while the percentage of caspase 3 positive cells was not affected by AA. PAS-staining showed that glycogen content in hepatocytes was not affected by AA. Serological examination revealed increase of lipid peroxidation in AA50mg group, while total protein concentration, protein thiol group level, as well as, paraoxonase 1 activity were not changed in AA-exposed animals. Stereological and immunohistochemical analyses of adult liver sections suggest increase of proliferation in AA50mg group, while increase of lipid peroxidation in serum of AA50mg group indicates oxidative stress induction.
La acrilamida (AA) es un químico ampliamente utilizado y un monómero importante en varios procesos industriales y de laboratorio. Además, la AA se forma durante el procesamiento de alimentos ricos en almidón a altas temperaturas. El objetivo de nuestro estudio fue examinar los efectos del tratamiento con AA subcrónica en el hígado de rata adulta utilizando métodos histológicos, estereológicos y bioquímicos. Se trataron ratas Wistar macho adultas con AA a dosis de 25 mg/kg p.v. y 50 mg/kg de peso corporal por tres semanas. El análisis estereológico mostró una disminución de la densidad del volumen del citoplasma de los hepatocitos y un aumento de la densidad del volumen de los núcleos de los hepatocitos y la relación nucleocitoplasmática en el grupo de 50 mg de AA. El análisis inmunohistoquímico de las secciones de hígado mostró que el tratamiento con 50 mg de AA aumentó el porcentaje de células positivas para PCNA, mientras que el porcentaje de células positivas para caspasa 3 no se vio afectado por AA. La tinción con PAS mostró que el contenido de glucógeno en los hepatocitos no se vio afectado por AA. El examen serológico reveló un aumento de la peroxidación de lípidos en el grupo de 50 mg de AA, mientras que la concentración de proteína total, el nivel del grupo tiol de proteína y la actividad de paraoxonasa 1 no cambiaron en los animales expuestos a AA. Los análisis estereológicos e inmunohistoquímicos de secciones de hígado adulto sugieren un aumento de la proliferación en el grupo AA50 mg, mientras que el aumento de la peroxidación lipídica en suero del grupo AA50 mg indica inducción de estrés oxidativo.
Subject(s)
Animals , Male , Rats , Acrylamide/administration & dosage , Liver/drug effects , Immunohistochemistry , Rats, Wistar , Proliferating Cell Nuclear AntigenABSTRACT
ObjectiveTo explore the effects of different treatment methods of "soothing liver, invigorating spleen, soothing liver and invigorating spleen, soothing liver first and then soothing liver and invigorating spleen, as well as invigorating spleen first and then soothing liver and invigorating spleen" on liver depression combined with liver injury in rats and their action mechanisms. MethodA six-week rat model of liver depression combined with liver injury was established by restraint stress and subcutaneous injection of carbon tetrachloride (CCl4, 5.89 g·kg-1, once every three days). At the same time, the drugs were given by gavage. Forty-eight male SD rats of clean grade were randomly divided into eight groups, namely the normal group, model group, bicyclol (0.2 g·kg-1) group, Sinisan (4.32 g·kg-1) group, Liu Junzitang (9.26 g·kg-1) group, Chaishao Liu Junzitang A (Chai A, soothing liver and invigorating spleen,13.57 g·kg-1) group, Chaishao Liu Junzitang B (Chai B, soothing liver first and then soothing liver and invigorating spleen, 13.57 g·kg-1) group, and Chaishao Liu Junzitang C (Chai C, invigorating spleen first and then soothing liver and invigorating spleen, 13.57 g·kg-1) group, with six rats in each group. The pathological changes in liver and colon tissues of each group were observed under light microscope and electron microscope. The serum biochemical indexes of the liver were detected using an automatic biochemical analyzer. The relative mRNA expression levels of Takeda G protein-coupled receptor 5 (TGR5) and intestinal mucosal zona occluden-1 (ZO-1), Occludin, and Claudin-1 in the liver and colon were detected by reverse-transcription polymerase chain reaction (RT-PCR). The positive expression rate of proliferating cell nuclear antigen (PCNA) in the colon was detected by immunohistochemistry. ResultCompared with normal group, the model group exhibited significantly elevated serum alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), and direct bilirubin (DBIL) (P<0.01), lowered TGR5 mRNA expression in liver tissue, up-regulated TGR5 mRNA expression in the colon tissue (P<0.05,P<0.01), and down-regulated ZO-1, Occludin, and tight junction protein-1 (Claudin-1) mRNA expression and PCNA in the colon tissue (P<0.01). Compared with the model group, bicyclol and Chai C remarkably decreased the levels of serum ALP, ALT, AST, TBIL, and DBIL (P<0.05,P<0.01), while Liu Junzitang, Chai A, Chai B, and Chai C significantly up-regulated the TGR5 mRNA expression in the liver and down-regulated its expression in the colon (P<0.01). Bicyclol, Chai A, Chai B, and Chai C enhanced the ZO-1 and Claudin-1 mRNA expression in the colon (P<0.05,P<0.01). Bicyclol, Sinisan, and Chai C increased PCNA expression (P<0.01). The comparison with the Chai C group showed that the TGR5 mRNA expression in the liver and ZO-1 mRNA expression in the colon of the bicyclol and Sinisan groups were lower, whereas the TGR5 mRNA expression in the colon was higher (P<0.01). However, the PCNA expression in the colon of the Liu Junzitang and Chai B groups declined significantly (P<0.05). ConclusionIn the presence of liver injury, invigorating spleen first helps to relieve the liver injury, and the efficacy of "spleen-invigorating" therapy in increasing the intestinal mucosal tight junction proteins and improving the gastrointestinal function is related to its activation of TGR5 to improve the intestinal mucosal barrier function, promote the renewal of intestinal stem cells, and drive the regeneration after injury.
ABSTRACT
Objective:To study the expression level and clinical significance of minichromosome maintenance protein 3(MCM3) in human gastric cancer.Methods:The expression levels of MCM3 mRNA and protein in gastric cancer and adjacent normal tissues were analyzed using public databases such as TCGA, CCLE and HPA. The clinicopathological features of 69 patients with gastric cancer were retrospectively analyzed, and the expression levels of MCM3 protein in tumor tissues and adjacent normal tissues were detected by immunohistochemical staining, and the relationship between it and clinicopathological features was analyzed. The interaction network of MCM3 proteins was analyzed using the STRING database.Results:The expression levels of MCM3 mRNA and protein in gastric cancer tissues were significantly higher than those in adjacent normal tissues ( P<0.05), and its high expression was correlated with the size of gastric cancer tumors ( P<0.05). In gastric cancer tissue, the expression of MCM3 was correlated with the expression level of proliferating cell nuclear antigen (PCNA) ( R=0.61, P<0.01), and there may be a protein-protein interaction. Conclusions:MCM3 plays an important role in gastric cancer by interacting with PCNA, and it is expected to become a new diagnostic and therapeutic target.
ABSTRACT
Objective:To analyze the expression level of minichromosome maintenance protein 6 (MCM6) in colon cancer tissues, the correlation between the expression level of MCM6 and the clinicopathological characteristics of colon cancer patients, and the correlation between MCM6 and PCNA expression.Methods:The expression levels of MCM6 in different tumor tissues were analyzed based on the Human Protein Atlas (HPA) database. The expression levels and correlations of MCM6 and PCNA in colon cancer tissues were analyzed based on The Cancer Genome Atlas (TCGA) database and immunohistochemical experiments. The correlation between MCM6 expression level and clinical characteristics of colon cancer patients was analyzed. The correlation between MCM6 and PCNA expression in colon cancer was analyzed based on TCGA database and Gene Expression Profile Interaction Analysis (GEPIA) database.Results:Bioinformatics analysis and immunohistochemical results confirmed that MCM6 was highly expressed in colon cancer tissues, and its expression level was correlated with the tumor stage of patients ( P=0.01). In colon cancer, the expression of MCM6 and PCNA was correlated with statistical significance ( P<0.05). Conclusions:MCM6 is highly expressed in colon cancer tissue and is related to the clinical characteristics of patients, suggesting that MCM6 can be used as a potential marker of colon cancer.
ABSTRACT
SUMMARY: GDM is linked with overexpression of inflammatory cytokines and increased oxidative stress, leading to endothelial dysfunction and vascular disorder. Weaimed to examine the expression of ADAMTS13 and PCNA in the placentas of gestational diabetes mellitus (GDM) patients to investigate the effects of hypoxia, induced by GDM, on proliferation and extracellular matrix formation in the maternal and fetal placenta cells. A total of 60 placentas were collected from pregnant women admitted to the obstetrics clinic. Thirty of them were diagnosed with GDM, and 30 of them were diagnosed with non-GDM patients. Samples were fixed in 10 % formaldehyde, after routine follow-up, embedded in paraffin wax. Sections of 5 µm were cut stained with Mayer Hematoxylin-Eosin, examined under a light microscope. Sections for immunohistochemical analysis were cut and processed for antigen retrievalin citrate solution. Sections were incubated with ADAMTS13 and PCNA primary antibodies, counterstained with hematoxylin, and evaluate under a light microscope. In histopathological examination, the non-diabetic placentas showed that decidua cells in the maternal region were polygonal with oval nuclei and organized in groups. In the GDM group, there were pyknosis and apoptotic changes in decidua cell nuclei. Vacuolar areas were observed in large cavities in maternal connective tissue. Inflammation and dilatation with congestion were observed in the blood vessels of the villus. In the GDM group, positive ADAMTS13 expression was observed in the decidua cells vascular endothelial cells, and surrounding connective tissue fibroblast cells. In the GDM group, a significant increase in PCNA expression was observed in decidua cells, connective tissue cells and endothelial cells. Functional changes in ADAMTS13 proteases and PCNA were thought to induce maternal and fetal complications by stimulating extracellular matrix development.
RESUMEN: La diabetes gestacional está relacionada con la sobreexpresión de citocinas inflamatorias y aumento del estrés oxidativo, lo que lleva a una disfunción endotelial y un trastorno vascular. Nuestro objetivo fue examinar la expresión de ADAMTS13 y PCNA en las placentas con diabetes mellitus gestacional (DMG) para investigar los efectos de la hipoxia inducida por DMG sobre la proliferación y formación de matriz extracelular en células placentarias maternas y fetales. Se recolectaron un total de 60 placentas de mujeres embarazadas ingresadas a la consulta de obstetricia. Treinta de ellas fueron diagnosticadas con DMG y 30 diagnosticadas sin DMG. Las muestras se fijaron en formaldehído al 10 %, y luego de un seguimiento de rutina, fueron embebidas en parafina. Se cortaron secciones de 5 µm teñidas con hematoxilina-eosina de Mayer, las que fueron examinadas bajo un microscopio óptico. Se cortaron y procesaron las secciones para el análisis inmunohistoquímico para la recuperación de antígeno en solución de citrato. Las secciones se incubaron con anticuerpos primarios ADAMTS13 y PCNA, se contratiñeron con hematoxilina y se evalua- ron con un microscopio óptico. En el examen histopatológico, las placentas no diabéticas mostraron que las células de la decidua en la región materna eran poligonales con núcleos ovalados y organizadas en grupos. En el grupo de DMG, se observó picnosis y cambios apoptóticos en los núcleos de las células de la decidua. Se observaron áreas vacuolares en el tejido conectivo materno. En los vasos sanguíneos de las vellosidades se observó inflamación y dilatación con congestión. En el grupo de DMG, se observó expresión positiva de ADAMTS13 en las células de la decidua, en las células endoteliales vasculares y en los fibroblastos del tejido conectivo circundante. En el grupo de DMG se observó un aumento significativo de la expresión de PCNA en células de la decidua, células de tejido conectivo y en las células endoteliales. Se considera que los cambios funcionales en las proteasas ADAMTS13 y PCNA inducen a complicaciones maternas y fetales al estimular el desarrollo de la matriz extracelular.
Subject(s)
Humans , Female , Pregnancy , Adult , Placenta/metabolism , Diabetes, Gestational/metabolism , Proliferating Cell Nuclear Antigen/metabolism , ADAMTS13 Protein/metabolismABSTRACT
Aim To explore the effects of EGCG on xenografts of ovarian cancer in nude mice and its possible mechanism. Methods Nude mice xenografts of ovarian cancer SK0V3 cells were established and divided into five groups after tumor formation, in which three groups were given EGCG (10, 30, 50 mg • k g-
ABSTRACT
Chronic hepatitis B virus (HBV) infection is a primary cause for liver cancer. And the main challenge of curing hepatitis B is the elimination of the stable covalently closed circular DNA (cccDNA) of the viral genome. The formation of HBV cccDNA requires the filling of single-stranded region and the ligation of nicks in relaxed circular DNA (rcDNA) strands. Previously, our group reported that proliferating cell nuclear antigen (PCNA) was involved in the formation of HBV cccDNA. However, the underlying mechanism of the conversion of HBV rcDNA to cccDNA is poorly understood. In the present study, we aim to explore the mechanism by which PCNA contributes to the conversion of HBV rcDNA to cccDNA. Our data showed that PCNA was involved in the process of HBV rcDNA repair. The knockout of PCNA by the CRISPR/Cas9 system remarkably blocked the conversion of HBV rcDNA to cccDNA, while the ectopic expression of PCNA could effectively rescue the event (P<0. 001). Knockout of PCNA significantly slowed down the conversion kinetics of HBV rcDNA to cccDNA (P<0. 01). Mechanically, the DNA binding domain of PCNA was required for the process of HBV rcDNA repair to cccDNA (P<0. 01). Thus, we conclude that PCNA confers the conversion of HBV rcDNA to cccDNA by its DNA binding domain. Clinically, PCNA might serve as a novel target for antiviral therapy.
ABSTRACT
Continuous industrial productivity and modern societies have resulted in excess artificial light. The altered circadian rhythm causes many diseases. During intrauterine life, the mother's maternal melatonin rhythm has a major role in influencing organ development. The aim of this study was to investigate the effect of maternal exposure to constant light on the structure and ultrastructure of neonatal skin. Twenty pregnant New Zealand rabbits were divided into two groups (n=10 each): control group (12-h light/dark) and constant light group (24-h light). Plasma maternal melatonin and corticosterone during pregnancy were determined. At the end of the experiment, the dorsal skin of the neonates of both groups was collected and prepared for histological, morphometric, and transmission electron microscopic study. Histological and morphometric results of skin of neonates from the constant light group revealed statistically significantly reduced epidermal thickness, decreased number of hair follicle, increased surface area of collagen, and decreased proliferating cell nuclear antigen (PCNA) positive cells. Ultrastructural examination showed wide intercellular spaces and disrupted desmosomal junctions in the epidermis. Earlier stages of hair follicles were also observed with indented shrunken nuclei, vacuolization, and swollen mitochondria. Dermal fibroblasts with dilated cisternae of rough endoplasmic reticulum containing electron-dense material were detected. Maternal melatonin was significantly reduced in the constant light group while maternal corticosterone showed no significant difference between groups. Therefore, normal maternal circadian rhythm is a key factor for the integrity of neonatal skin structure.
Subject(s)
Animals , Female , Pregnancy , Rabbits , Skin , Melatonin , Circadian Rhythm , Microscopy, Electron, Transmission , EpidermisABSTRACT
In the current study, the histological structure of the gallbladder of Alburnus tarichi (Güldenstädt, 1814) was investigated. Hematoxylin and eosin were used to stain the histological sections for routine examinations, in addition to using periodic acid Schiff (PAS) for the neutral mucins, aldehyde fuchsin (AF) for the sulphated mucins, and Alcian blue (AB; pH: 2.5) for the acidic mucins. In addition, proliferating cell nuclear antigen (PCNA) immune-staining was performed for the detection of dividing cells among the epithelium. The gallbladder of A. tarichi was composed of mucosa, muscularis, and serosa or adventitia layers. The mucosa covering the wavy pleomorphic folds was made up of tall columnar epithelium and a lamina propria. The apical surface of the epithelial cells was lined by continuous short microvilli. On the epithelium, the luminal surface was remarkably stained with PAS, AF, and AB. Slight to moderate staining was observed on the epithelial cells in the apical zone with PAS. The cytoplasm of epithelial cells were stained in a slight manner with AF. No goblet cells were observed among the epithelium. According to the PCNA immune-staining, some epithelial cells were observed to proliferate. The lamina propria consisted of loose connective tissue that contained fibrocytes, collagen and elastic fibers, capillaries, and small blood vessels. The muscularis layer displayed muscle fibers that were circular, smooth, and surrounded by collagen fibers. The subserosal and serosal or adventitial layers had typical morphology to those of other fish and vertebrates.
En este estudio, se investigó la estructura histológica de la vesícula biliar de Alburnus tarichi (Güldenstädt, 1814). Las secciones histológicas se tiñeron con Hematoxilina-Eosina para los exámenes de rutina, además de usar el ácido periódico de Schiff (PAS) para las mucinas neutras, aldehído fucsina (FA) para las mucinas sulfatadas y azul alcián (AB; pH: 2,5) para las mucinas ácidas. Además, se realizó una tinción inmune de antígeno nuclear de células proliferativas (PCNA) para la detección de células en división entre el epitelio. La vesícula biliar de A. tarichi estaba compuesta de capas, mucosa, muscular y serosa o adventicia. La mucosa que cubría los pliegues pleomórficos ondulados estaba formada por un epitelio columnar alto y una lámina propia. Se observó una superficie apical de las células epiteliales revestida por microvellosidades cortas y continuas. En el epitelio se observó una tinción importante de la superficie luminal teñida con PAS, FA y AB. Se observó una tinción leve a moderada en las células epiteliales en la zona apical con PAS. El citoplasma de las células epiteliales se tiñó ligeramente con FA. No se observaron células caliciformes entre el epitelio. Según la tinción de PCNA, se observó que proliferaban algunas células epiteliales. La lámina propia consistía en tejido conectivo laxo que contenía fibrocitos, colágeno y fibras elásticas, capilares y pequeños vasos sanguíneos. La capa muscular mostraba fibras musculares circulares, lisas y rodeadas de fibras de colágeno. Las capas subserosas y serosas o adventicias tenían una morfología típica a la de otros peces y vertebrados.
Subject(s)
Animals , Cyprinidae/anatomy & histology , Gallbladder/anatomy & histology , Proliferating Cell Nuclear AntigenABSTRACT
This study aimed to assess the effects of Ginkgo Biloba Extract and Troxerutin on the hippocampus of induced diabetes mellitus in adult albino rats using histological methods.50 adult male albino rats were divided into three groups; Group I (Control); Group II (diabetic): subdivided into Subgroup IIa (T1DM)), Subgroup IIb (T1DM+GBE), Subgroup IIc (T1DM+ troxerutin); Group III: subdivided into Subgroup IIIa (GBE) and Subgroup IIIb (troxerutin). The brain was removed and the cerebral hemisphere was coronally cut at the hippocampal level and used for light microscopic study (H&E staining and PCNA immunostaining). There was a statistically insignificant improvement in animal weights in subgroup IIb and subgroup IIc. Subgroup IIb showed a statistically significant reduction of blood glucose levels while the subgroup IIc showed insignificant reduction of blood glucose levels. Diabetes disturbed the light microscopic structure of the hippocampus. In subgroup IIb and subgroup IIc the hippocampus retained an apparently normal appearance and the stratum pyramidale exhibited the pyramidal cells with rounded vesicular nuclei and acidophilic cytoplasm. Diabetic hippocampal sections revealed negative PCNA immunoreactivity inall layers of DG. In subgroup IIb and subgroup IIc, hippocampal sections showed positive immunoreactivity
ABSTRACT
Ethephon (Ethrel®) is an ethylene-based plant growth regulator that used in agriculture and it has direct and indirect effects on human health, direct effect via its inhalation during usage in agriculture and indirect effect through the diet (Fruits and vegetables) that is sprayed with it. The current study aimed to examine the possible modifying effects of costus (Saussurea lappa) root aqueous extract against Ethephon induced liver toxicity, injury, DNA fragmentation and PCNA alterations in male rats. Fifty adult male rats were divided into 5 groups (1st, control; 2nd, Costus; 3rd, Ethephon; 4th, Post treated Ethephon with costus; 5th, self-healing Ethephon). Current results revealed that; a significant increase in aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), liver injury, DNA damage and PCNA expressions in Ethephon group when compared with control group. In contrast; a significant decrease in albumin and total proteins in Ethephon group when compared with control group. Treatment of rats with costus after Ethephon improved these alterations as compared with Ethephon self-healing. So, it could be concluded that costus root extract worth to be considered as a natural substance for ameliorating the hepatic toxicity induced by plant growth regulator Ethephon.
Subject(s)
Animals , Male , Rats , Plant Growth Regulators/agonists , Plant Extracts/analysis , Costus/adverse effects , Chemical and Drug Induced Liver Injury , DNA Damage , Inhalation , Proliferating Cell Nuclear Antigen , Agriculture/classification , Liver/abnormalitiesABSTRACT
Abstract Objective To analyze the effect of thalidomide on the progression of endometriotic lesions experimentally induced in rats and to characterize the pattern of cell proliferation by immunohistochemical Proliferating Cell Nuclear Antigen (PCNA) labeling of eutopic and ectopic endometrium. Methods Fifteen female Wistar rats underwent laparotomy for endometriosis induction by resection of one uterine horn, isolation of the endometrium and fixation of a tissue segment to the pelvic peritoneum. Four weeks after, the animals were divided into 3 groups: control (I), 10mg/kg/day (II) and 1mg/kg/day (III) intraperitoneal thalidomide for 10 days. The lesion was excised together with the opposite uterine horn for endometrial gland and stroma analysis. Eutopic and ectopic endometrial tissue was submitted to immunohistochemistry for analysis of cell proliferation by PCNA labeling and the cell proliferation index (CPI) was calculated as the number of labeled cells per 1,000 cells. Results Group I showed a mean CPI of 0.248 ± 0.0513 in the gland and of 0.178 ± 0.046 in the stroma. In contrast, Groups II and III showed a significantly lower CPI, that is, 0.088 ± 0.009 and 0.080 ± 0.021 for the gland (p < 0.001) and 0.0945 ± 0.0066 and 0.075 ± 0.018 for the stroma (p < 0.001), respectively. Also, the mean lesion area of Group I was 69.2mm2, a significantly higher value compared with Group II (49.4mm2, p = 0.023) and Group III (48.6mm2, p = 0.006). No significant difference was observed between Groups II and III. Conclusion Thalidomide proved to be effective in reducing the lesion area and CPI of the experimental endometriosis implants both at the dose of 1mg/kg/day and at the dose of 10 mg/kg/day.
Resumo Objetivo Analisar o efeito da talidomida na progressão de lesões endometrióticas induzidas experimentalmente em ratas e caracterizar o padrão de proliferação celular pela marcação imunohistoquímica de Antígeno Nuclear de Célula Proliferativa (PCNA) no endométrio eutópico e ectópico. Métodos Quinze ratas Wistar foram submetidas a laparotomia para indução de endometriose por ressecção de um corno uterino, isolamento do endométrio e fixação de um segmento do tecido ao peritônio pélvico. Após quatro semanas, os animais foram divididos em 3 grupos: controle (I), 10 mg/kg/dia (II) e 1 mg/kg/dia (III) de talidomida intraperitoneal por um período de 10 dias. As lesões foram resseccionadas juntamente com o corno uterino oposto para análise da glândula endometrial e do estroma. O tecido endometrial eutópico e ectópico foi submetido à imunohistoquímica para análise da proliferação celular por marcação com PCNA e o índice de proliferação celular (CPI) foi calculado como o número de células marcadas por 1.000 células. Resultados O grupo I apresentou média de CPI de 0,248 ± 0,0513 na glândula e de 0,178 ± 0,046 no estroma. Em contraste, os grupos II e III apresentaram CPI significativamentemenor, isto é, 0,088 ± 0,009 e 0,080 ± 0,021 para a glândula (p < 0,001) e 0,0945 ± 0,0066 e 0,075 ± 0,018 para o estroma (p < 0,001), respectivamente. Além disso, a área de lesãomédia do Grupo I foi de 69,2mm2, valor significativamentemaior em relação ao Grupo II (49,4mm2, p = 0,023) e Grupo III (48,6mm2, p = 0,006). Não houve diferença estatisticamente significante entre os Grupos II e III. Conclusão A talidomida mostrou-se eficaz na redução da área da lesão e CPI dos implantes de endometriose experimental tanto na dose de 1mg/kg/dia quanto na dose de 10 mg/kg/dia.
Subject(s)
Humans , Animals , Female , Thalidomide/pharmacology , Angiogenesis Inhibitors/pharmacology , Disease Models, Animal , Endometriosis/pathology , Endometrium/pathology , Biomarkers/analysis , Rats, Wistar , Proliferating Cell Nuclear Antigen/analysis , Cell Proliferation/drug effects , Dose-Response Relationship, DrugABSTRACT
;Aim To investigate the role of periostin in PDGF-induced proliferation and extracellular matrix accumulation of mouse mesangial cells. Methods The effects of PDGF on periostin protein, cell proliferation and extracellular matrix accumulation were detected. The cells were collected at 0, 2,4, 6, 12 h after stimulation with PDGF(10 (ig • L " 1 ) to detect the expression of periostin, PCNA, FN and TGF-ßl by Western blot. The silencing effect of sh-periostin vector on periostin protein in mouse mesangial cells was identified by Western blot. Cells were randomly divided into control group, PDGF group, PDGF + sh-nc group and PDGF + sh-periostin group to detect the role of periostin in PDGF-induced proliferation and extracellular matrix accumulation of mouse mesangial cells. Results PDGF could elevate periostin protein expression. Western blot result showed that periostin protein expression in PDGF-stimulated groups was significantly higher than that in Oh group, which was consistent with the result of immunofluorescence. Positive expression of periostin was located in cytoplasm. Western blot result showed that PCNA, FN and TGF-ßl protein in PDGF-stimulated groups increased as compared with Oh group. shRNA vector aimed at periostin (sh-periostin vector) could partially reverse PDGF-induced mesangial cell proliferation and extracellular matrix expression. PCNA, Fn and TGF-ßl expressions were attenuated significantly. Conclusions PDGF can enhance periostin protein expression and increase mouse mesangial cell proliferation and extracellular matrix accumulation. Periostin shRNA vector can partially reverse PDGF-induced mesangial cell proliferation and extracellular matrix generation.
ABSTRACT
Objective: To clarify the role of discs large homolog 5 (DLG5) in the diagnosis and prognosis of renal clear cell carcinoma (ccRCC) by studying the expression of DLG5 in the clinical tissue of ccRCC. Methods: DLG5 expression in ccRCC tissues and normal renal tissue was measured by using the Cancer Genome Atlas (TCGA) Database and Gene Expression Omnibus (GEO) Database. The correlation of DLG5 expression with the prognosis and clinicopathological parameters of ccRCC was analyzed by LinkedOmics and GEPIA. The interactions protein network of DLG5 was predicted by String-DB. Results: Compared with that of the normal kidney tissues, DLG5 expression was significantly upregulated in the ccRCC tissues (P<0.05). The high DLG5 expression was an independent prognostic factor affecting the overall survival rate of ccRCC based on TCGA (P=2.997×10-5). Furthermore, DLG5 expression had a positive correlation with the TNM stage, ethnicity, tumor purity, year-to-birth, PCNA (P=1.1×10-6, R=0.19), and RAD1 (P=5.7×10-6, R=0.18) expression of ccRCC. Conclusion: In ccRCC, the high expression of DLG5 is a significant index of poor prognosis. Therefore, DLG5 can be a potential biomarker for predicting the metastasis, recurrence and prognosis of the patients.
ABSTRACT
Objective To investigate the effect of Sargentodoxa cuneata extracts combined with 5-fluorouracil on the growth inhibition of human hepatoma HepG2 cells, and explore its mechanism. Methods HepG2 cells were cultured in vitro and treated with different concentrations of S. cuneata extracts. The effect of S. cuneata extracts on cell growth inhibition was detected by MTT assay, and the subsequent experimental concentration was selected. HepG2 cells were randomly divided into four groups: control group, S. cuneata extracts group (40 mg/L), 5-fluorouracil (10 μmol/L) group, and combination group (S. cuneata extracts 40 mg/L + 5-fluorouracil 10 μmol/L). Cell proliferation inhibition rate was detected by MTT assay and cell cycle was detected by flow cytometry. The expression levels of nuclear antigen PCNA, Cyclin D1, and cell cycle-dependent protein kinase CDK4 were detected by Western blotting. Results The results of MTT assay showed that S. cuneata extracts inhibited the proliferation of HepG2 cells in a concentration-dependent manner (P < 0.05), and S. cuneata extracts with a concentration of 40 mg/L was selected for subsequent experiments. Compared with the control group, the cell proliferation inhibition rate of the S. cuneata extracts group and the 5-fluorouracil group was significantly increased, the proportion of G0/G1 phase cells was significantly increased, and the proportion of cells in the S phase and G2/M phase was significantly decreased (P < 0.05). The protein expression levels of PCNA, Cyclin D1, and CDK4 in the cells were significantly decreased (P < 0.05). Compared with the 5-fluorouracil group, the combination group significantly inhibited the proliferation of HepG2 cells, blocked the cell cycle, and inhibited the protein expression of PCNA, Cyclin D1, and CDK4 in the cells (P < 0.05). Conclusion S. cuneata extracts combined with 5-fluorouracil enhances the growth inhibition of hepatocellular carcinoma HepG2 cells, and its mechanism may be related to the inhibition of the protein expression of PCNA, Cyclin D1, and CDK4 related to the expression levels of PCNA, Cyclin D1, and CDK4 in the inhibited cells.
ABSTRACT
Purpose To study the expression and the clinical significance of β3GnT8,MMP-2,PCNA in gastric cancer tissue. Methods The histological chips of paraffin specimens of gastric cancer were prepared,and immunohistochemical methods were used to detect the expression of β3GnT8,MMP-2 and PCNA in gastric cancer and adjacent tissue. Results The expression of β3GnT8 and MMP-2,PCNA in gastric cancer was higher than that in adjacent tissue (P = 0. 001) . The expression of β3GnT8 and MMP-2 was significantly positively correlated with the clinical stage (P = 0. 001) ,depth of invasion (P = 0. 011) and lymph node metastasis (P = 0. 003) of gastric cancer. The expression of β3GnT8 was significantly positively correlated with that of MMP-2 (r = 0. 703,P = 0. 001) and PCNA (r = 0. 231,P = 0. 024) . The overall survival time of the β3GnT8 positive expression group was significantly shorter than that of the negative expression group(χ2 = 3. 957,P = 0. 047) . Conclusion The expression of β3GnT8 is increased in gastric cancer. β3GnT8 can promote the invasion and metastasis of gastric cancer and is associated with poor prognosis in patients with gastric cancer.
ABSTRACT
Objective: To study the apoptosis mechanism of microwave ablation (MA) in tongue cancer Cal-27 cell transplanted tumor in nude mice. Methods: 20 nude mice with Cal-27 cell subcutaneously transplanted tumor were randomly divided into blank control group, 50 ℃ 1 min MA group, 50 ℃ 2 min MA group and 55 ℃ 2 min MA group (n = 5) respecitvely, and were treated with MA with indiated perameters respectively. The apoptosis of tumor cells was observed by HE staining. The expression of Caspase-3 and PCNA protein was detected by immunohistochemical SP method. Results: HE staining showed that microwave ablation induced apoptosis of the tumor cells. Immunohistochemical results showed that the expression of Caspase-3 protein was significantly up-regulated in the microwave ablation groups compared with the control group (P < 0. 05), and the expression of PCNA protein was significantly decreased (P < 0. 05) . In microwave ablation 55 ℃ 2 min group, the expression of Caspase-3 protein was increased more (P <0. 01), and the expression of PCNA protein was decreased more (P < 0. 01) . Conclusion: Microwave ablation may inhibite the proliferation of tongue cancer cells by inducing apoptosis of the cells. The mechanism may be related to the up-regulation of Caspase-3 signaling pathway and down-regulation of PCNA signaling pathway.