• Title/Summary/Keyword: PCNA (proliferating cell nuclear antigen)

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Albizzia julibrissin Suppresses Testosterone-induced Benign Prostatic Hyperplasia by Regulating 5α-Reductase Type 2 - Androgen Receptor Pathway

  • Hong, Geum-Lan;Kim, Hyun-Tae;Park, Se-Ra;Lee, Na-Hyun;Ryu, Kyung-A;Kim, Tae-Won;Song, Gyu-Yong;Jung, Ju-Young
    • Natural Product Sciences
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    • v.25 no.3
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    • pp.200-207
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    • 2019
  • Albizzia julibrissin (AJ) is an herbal medicine that shows low toxicity, promotes promoting blood circulation and mitigates the inflammation and has mild side effects. Benign prostate hyperplasia (BPH) is one of the most common diseases that occurs in older males and often results in lower urinary tract symptoms. This study was conducted to evaluate the protective effect of AJ against BPH using LNCaP cells and Sprague Dawley rats treated with testosterone. Treatment with AJ extract reduced the expression of androgen receptor (AR) and prostate-specific antigen (PSA) in vitro. In vivo, rats were divided into 6 groups: 1 (Normal Control); 2 (Testosterone propionate (TP) alone); 3 (TP + finasteride); 4 (TP + AJ 10 mg/kg); 5 (TP + AJ 50 mg/kg); 6 (TP + AJ 300 mg/kg). The groups treated with AJ showed reduced the relative prostate weights and BPH-related proteins were altered, with decreased AR, PSA and proliferating cell nuclear antigen (PCNA) observed by western blot. Histopathological analysis revealed the therapeutic effect of AJ, with a decreased thickness of epithelial cells and reduced level of PCNA and $5{\alpha}$-reductase type 2. These results suggest that AJ extract could ameliorate testosterone-induced benign prostatic hyperplasia.

Expression patterns of proliferating cell nuclear antigen in the small intestine of mice infected with Metagonimus yokogawai and Metagonimus Miyata type (요코가와흡충 및 미야타흡충 감염 마우스 소장의 PCNA발현 양상)

  • Yu, Jae-Ran;Myeong, Na-Hye;Chae, Jong-Il
    • Parasites, Hosts and Diseases
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    • v.35 no.4
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    • pp.239-244
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    • 1997
  • Genus Metasonimus has been a subject of taxonomic debates for several years. In morphological aspects, M. vokogawci has been thought to have three subtypes, namely Yokogawa type (M. yokogawai in strict sense) , Miyata type and Koga type. But dif- ferences in the intestinal pathology induced by these subtypes have not been studied yet. In this study we compared the pathological reactions inducted by M. yokoBnwai and Metcgonimus Miyata type using proliferating cell nuclear antigen (PCNA) index. Metacercariae (Mc) of M. yokogcuani were collected by artificial digestion of Plecoslossw oLtiveLis and Mc of Metngonimus Miyata type were collected from Zacco pEntvpus. Three hundreds Mc of each species were infected orally to ICR mice. The mice were sacrificed at 3, 6. 10. 16 and 23 days after infection and the small intestines were resected into three portions (proximal. middle, and distal) . Immunohistochemical staining for PCNA was done using PC-10 (DAKO-PCNA, CA, USA) The PCNA indices in M. Wokosnwci infected group on the 6th and 23rd day after infection were lower than in the control and Miyata type infect ed groups (p < 0.05) from all of the three intestinal regions. On the other hand, the control group and Ifetngonimus Miyata type infected group did not make any differences in PCNA indices. The villus/crypt WIC) ratio was also decreased significantly in M. Wokogcwni infected mice but not in Metagonimus Miyata type infected ones. It is suggested that M. wokogawci induce villous atrophy through a decrease in the cell proliferation at the crypt. The results of this study suggested that M. WokoBnwai induce more serious intestinal pathology than Metcsonimuf Miyata type.

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Busulfan Inhibits PCNA Expression but Induces The Expression of pRB

  • 천영신;주학진;권득남;김진희
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.21-21
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    • 2001
  • 일반적으로 세포는 방사능이나 항암제 등의 자극에 의해 DNA가 손상받았을 때 DNA를 합성하기 전, DNA변이를 복구하기 위해 cell cycle을 정지시키게 된다. pRB(retinoblatoma protein)는 이러한 cell cycle의 조절기작에서 중요한 역할을 담당하는 것으로 알려져 있다. G1기에서 S기로 진행하는 것을 조절하는 단백질인 pRB 은 E2F(cell cycle transcription factor)와 상호작용하여 cell cycle 진행에 필요한 전사활성을 억제, PCNA (proliferating cell nuclear antigen)의 합성을 저해한다. 또한, E2F와 결합된 pRB는 apoptosis를 제어하는 유전자를 조절하는 것으로 알려져 있다. Cell cycle에 영향을 미치는 항암제의 일종인 busulfan을 처리하면, 정소 내에 존재하는 대부분의 생식세포들은 사멸되고 spermatogonia만 남는 것으로 알려져 있다. 그러나 그 기작에 대해서는 자세히 연구된 바가 없다. 본 연구에서는 busulfan처리시 spermatogonial stem cell이 어떤 기작에 의해 손상받지 않고 유지되는지를 알아보고자 실험을 수행하였다. Busulfan을 처리한 마우스 (항암제 투여 후 5주)와 정상적인 13주령의 마우스의 정소로부터 각각 세포를 분리하였다. LSC (laser scanning cytometry)를 이용하여 처리군(busulfan treated mice)과 대조군(normal mature mice)에 대해 각각 DNA함량을 비교ㆍ분석한 결과 G0/G1(2N)에 머물러 있는 세포비율이 처리군에서 현저하게 증가했다 (79.3$\pm$5.5%:8.1$\pm$1.3%). Cell cycle의 G1/S check point인 pRB와 PCNA 발현을 Western blot과 면역조직학적인 방법(immunohisto-chemistry)을 이용하여 조사하였다 PCNA는 대조군과 비교해, 처리군에서 매우 낮은 수준으로 발현되었다. 면역염색된 정소단면을 살펴보면, 대조군에서는 모든 세정관에서 PCNA를 발현하는 세포가 높은 비율로 검출되었고, 처리군에서는 소수의 세정관에서 세포들이 낮은 수준으로 검출되었다. 반면에, pRB의 경우 PCNA와는 상반된 결과를 나타내어, 대조군에서는 거의 발현이 되지 않는 반면, 처리군에서는 대부분의 세정관내, 기저막을 따라 위치한 세포들에서 발현되었다. 이상의 결과는 busulfan에 의해 pRB의 인산화가 억제, pRB 와 결합된 E2F는 전사 활성이 억제되어, PCNA 합성을 저해하는 것으로 설명되어질 수 있다. 결론적으로, 인산화가 억제된 pRB (underphosphorylated RB protein)이 quiescent spermatogonial stem cell에서만 특이하게 발현하는 단백질이며, 이러한 pRB의 발현은 apoptosis를 제어하는 역할을 담당해 busulfan처리에 의해 손상받지 않고 남아있는 것으로 시사된다.

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Coexpression of PCNA and p21 for DNA repair in small intestinal crypt cells of mouse with 60Co γ-rays irradiation (방사선을 조사한 마우스의 소장 음와세포에서 DNA 수복을 위한 PCNA와 p21의 발현 양상)

  • Hong, Suji;Hwang, Insun;Ahn, Meejung;Shin, Taekyun;Joo, Hong-gu;Park, HyunJeong;Jee, Youngheun
    • Korean Journal of Veterinary Research
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    • v.45 no.4
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    • pp.457-464
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    • 2005
  • The irradiation of radioactive ${\gamma}-ray$ induces apoptosis of radiosensitive organs for homeostasis. In this study, we investigated the repair mechanisms for homeostasis in the small intestine after cell damage by $^{60}Co\;{\gamma}-ray$ irradiation. The apoptosis was most frequently observed in the crypt cells of the small intestine after four and six hours by radioactive ${\gamma}-ray$ irradiation, and the frequency of apoptosis was proportional to the amount of irradiation. Also, the number of apoptotic cells was coincident with expression pattern of p53. Interestingly, PCNA (proliferating cell nuclear antigen) which is engaged in DNA replication and repair was expressed in apoptotic cells of small intestinal crypts. Also, it was observed that cell-cycle regulator p21 which is known to induce cell-cycle arrest is co-expressed in the same apoptotic cells of irradiated small intestinal crypt cells. These findings suggest that the co-expression of PCNA and p21 proteins, which may lead to resistance to DNA damage through cell-cycle arrest is closely associated with repair of damaged gastrointestinal cells after ${\gamma}-ray$ irradiation.

The Effect of Irradiation on the Expression of PCNA and Apoptosis Induction during the DMBA Induced Carcinogenesis in Hamster Buccal Pouch (DMBA 유도 햄스터 협낭암 발생과정에서 방사선 조사가 증식세포핵항원(PCNA)의 발현과 apoptosis 유발에 미치는 영향)

  • Jeon In-Seong;Heo Min-Suk;Choi Hang-Moon;Lee Sam-Sun;Choi Soon-Chul
    • Imaging Science in Dentistry
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    • v.30 no.3
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    • pp.207-216
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    • 2000
  • Purpose : This study was carried out to investigate the effect of irradiation on the expression of proliferating cell nuclear antigen (PCNA) and apoptosis induction during the carcinogenesis in hamster buccal pouch. Materials and methods: Three months old Syrian golden hamsters were divided into control and 2 experimental groups. Hamsters in control group were left untreated on buccal pouchs. Twenty four hamsters were treated with 0.5% DMBA tri-weekly on the right buccal pouch. Forty eight hamsters were treated with 0.5% DMBA tri-weekly and irradiated with the dose of 5 Gy and 10 Gy at 6, 9, 12, 15 weeks after DMBA application. Resected buccal pouches were sectioned and examined for potential expression pattern of PCNA and apoptosis. Results : The PCNA index was increased with the stages of buccal pouch epithelium carcinogenesis except the hyperplasia stage in control group (p<0.05). The irradiation did not effect on the PCNA index in the dysplasia and the carcinoma in situ stage, but in the hyperplasia stage, the PCNA index was increased with 10 Gy radiation and decreased in the carcinoma stage (p<0.05). The apoptotic index was significantly decreased from the carcinoma in situ stage and the lowest in the carcinoma stage. The apoptotic index was significantly decreased in the hyperplasia and dysplasia stage with the 5 Gy irradiation and significantly increased only in the carcinoma stage with the 10 Gy irradiation (p<0.05). Conclusion: The PCNA and apoptotic index were varied according to the irradiation period and dosage in each carcinogenesis stage.

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Histopathological Observation and Flow Cytometry Analysis of Testicular Atrophy Induced by 2-Bromopropane On the Sprague-Dawley Rat (2-Bromopropane에 의한 유발된 Sprague-Dawley 랫트의 고환위축의 병리학적 관찰 및 Flow Cytometry를 이용한 검사)

  • 손화영;강부현;조성환;차신우;노정구
    • Toxicological Research
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    • v.14 no.2
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    • pp.143-149
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    • 1998
  • This study was carried out to evaluate the testicular toxicity of 2-bromopropane (2-BP), which recently caused occupational intoxication on the reproductive and hematopoietic system in Koreans, using light microscopy, immunohistochemistry and flow cytometry. 10 weeks old male Sprague-Dawley (SD) rats were treated with 0.5 g/㎏/day of 2-BP orally for 8 consecutive weeks. The testes of the rats were vascularly perfused with Karnovsky's solution or immersed in Bouin's solution, embedded in plastic and evaluated with light microscopy. And relative proportions of haploid, diploid, and tetra-ploid states of DNA ploidy in the testicular cell suspensions of the SD rats were examined by flow cytometry. 2-BP induced severe testicular atrophy, depletion and degeneration of spermatogonia, spermatocytes, and spermatids and mild hyperplasia of Leydig cells without significant morphological changes. The Leydig cell hyperplasia was confirmed by immunohistochemistry using proliferating cell nuclear antigen (PCNA). The immunopositive cells against PCNA were observed in the nuclei oj some interstitial cells. Relative proportions of haploid states of DNA ploidy decreased in the atrophic testicular cell suspensions comparing with those of the control. In conclusion, 2-BP induced testicular atrophy with Leydig cell hyperplasia as examined by histopathology, immunohistochemistry and DNA flow cytometry.

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Cytokeratin 15 is an Effective Indicator for Progression and Malignancy of Esophageal Squamous Cell Carcinomas

  • Shen, Yu-Hong;Xu, Cui-Ping;Shi, Zhi-Meng;Zhang, Yan-Jiao;Qiao, Ya-Guang;Zhao, He-Ping
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.9
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    • pp.4217-4222
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    • 2016
  • Purpose: To compare the expression level of CK 15 in normal esophageal and esophageal squamous-cell carcinoma (ESCC) tissues and analyse possible functions of CK15 in occurrence and development. Materials and Methods: Immunohistochemistry was used to compare CK14, CK15 and proliferating cell nuclear antigen (PCNA) expression levels in ESCCs. Expression level of CK15 was also assessed by Western blotting. In addition, levels of CK15, cytokeratin 19 fragment antigen 21-1 (CYFRA21-1) and PCNA were detected in serum by enzymelinked immunosorbent assay (ELISA) and chemiluminescence methods. Relationships between clinicopathological parameters and CK14 and CK15 expression were then analyzed. Results: According to immunohistochemistry, in esophageal and intraepithelial neoplasia (SIN) tissues, the expression of CK14, CK15 and PCNA localized to basal layer of the epithelium. CK14 and CK15 levels were higher in normal esophageal squamous epithelial tissue than in SIN and ESCC, and greater in highly differentiated than poorly differentiated carcinoma tissue. By Western blotting, we found more pronounced expression of CK15 in normal esophageal tissue, compared with carcinoma tissue. The specificity of changed CK15 and CYFRA21-1 expression was respectively 90.0% and 96.7% in serum of ESCC patients. Joint detection could improve the sensitivity of esophageal carcinoma diagnosis. Relationships between CK14, CK15 expression and clinical parameters were not statistically significant (P>0.05). Postoperative survival in patients of CK14, CK15 positive expression was longer than with negative expression ($x^2=4.35$, P=0.037; $x^2=9.852$, P=0.002). Conclusions: CK15 expression decreased in esophageal squamous cell carcinoma tissue and serum of esophageal squamous carcinoma patients. We infer that CK15 may play an important role for the occurrence and development of esophageal squamous-cell carcinoma. In the future, CK15 may be used for the diagnosis, treatment and prognostic evaluation of esophageal squamous-cell carcinoma.

The Effect of Bifidobacteria and Various Oligosaccharides Consumption on the Risk of Colon Cancer in Rats

  • Khil, Jin-Mo
    • Nutritional Sciences
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    • v.8 no.4
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    • pp.219-225
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    • 2005
  • This study examined the effect of viable bifidobacteria and non-digestible carbohydrates on the cecal pH, colonic neoplastic lesion (aberrant crypt) and proliferating cell nuclear antigen (PCNA) labeling index in carcinogen-treated mts. Animals received s.c. injection of dimethylhydrazine (DMH) (15 mg/kg body weight) twice 3 days apart. Three days after the second carcinogen administration, the treatments were begun. 1he treatments were basal diet (AIN-76) with skim milk (Basal/skim), or the following diets with daily gavage of $10^8$ bifidobacteria: basal (Basal/bifido), $2\%$ fructo-oligosaccharide (FOS/bifido), $2\%$ soybean oligosaccharide (SBO/bifido), $2\%$ wheat bran oligosaccharide (WBO/bifido) and $8.4\%$ wheat bran (WB/bifido). After 4 weeks of treatment, cecal pH was measured using a pH probe. The number of aberrant crypt (AC), aberrant crypt foci (ACF) and crypt multiplicity were enumerated and colonic PCNA labeling index was determined using immunohistochemistry. Cecal pH was significantly reduced in SBO/bifido and FOS/bifido groups compared to control group. However, there were no significant differences in either number of AC or rates of cell proliferation as shown by PCNA labeling index among the groups, although mts fed FOS/bifido reduced the numbers of ACF compared to Basal/skim group. The SBO/bifido group did not reduce the number of ACF or PCNA labeling index. Also, other oligosaccharides did not reduce the risk of colon cancer compared to control group. The concomitant reduction of cecal pH and number of ACF suggest that the combination of bifidobacteria and FOS may reduce the risk of colon cancer.

AN IMMUNOHISTOCHEMICAL STUDY ON MANDIBULAR CONDYLE IN WEANLING RAT AFTER MASSETER MUSCLE RESECTION (이유백서에서 교근절제후 하악과두의 면역조직화학적 연구)

  • Choi, Nam-Ki;Choi, Hong-Ran;Yang, Kyu-Ho
    • Journal of the korean academy of Pediatric Dentistry
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    • v.23 no.2
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    • pp.559-574
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    • 1996
  • The purpose of this study was to determine whether there was any correlation between temporomandibular joint dysfunction and structure of the mandibular condyle. Weanling rats had their masseter muscles resected and immunohistochemical findings were observed with a light microscope. The results obtained were as follows : 1. The condylar cartilage region was divided into articular, proliferating, cartilage cell and hypertrophic cell layers according to cell morphology. 2. In light microscopic views, the proliferating and cartilage cell layers of the experimental group decreased gradually and at the 8th week significantly. 3. In immunohistochemical staining for type I and II collagen, a reaction was detected in the lower part of proliferating cell and cartilage cell layers. In the cartilage cell layers, a stronger cellular reaction was present. Immunohistochemical staining for type II collagen reacted more strongly than that of type I collagen. 4. In immunohistochemical staining for proteoglycan, the staining of the experimental group resembled the control group and gradually showed a weak reaction. The proliferating and cartilage cell layers reacted more strongly than the hypertrophic cell layer. 5. In immunohistochemical staining for proliferating cell nuclear antigen(PCNA), the strong reaction was detected in the nucleus of the proliferating cell layer both in control and experimental groups. But the thickness of the proliferating layer decreased in experimental group, consequently the reaction of the experimental group was reduced more than that of the control group.

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