• 제목/요약/키워드: immunohistochemical methods

검색결과 765건 처리시간 0.031초

유근피(楡根皮) 약침이 lipopolysaccharide 유발 류마티스 관절염 모델에서 MIF 활성 억제에 미치는 영향 (The Effectiveness of Ulmus Davidiana Planch Herbal Acupuncture to Inhibit MIF Activation on Lipopolysaccharide-induced Rheumatoid Arthritis Model)

  • 변혁;박인식;조현석;김갑성;이승덕
    • Journal of Acupuncture Research
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    • 제23권6호
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    • pp.117-132
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    • 2006
  • Objectives : The purpose of this study is to investigate the effect of Ulmus davidiana Planch herbal acupuncture solution in LPS-stimulated RAW 264.7 cells and mouse knee joints, perfom1ed several experimental items: those are MIF mRNA, MIF, $TNF-{\alpha}$, $NF-{\kappa}B$ p65, iNOS mRNA, iNOS, NO, synovial hyperplasia, angiogenesis and fibrosis. Methods : In order to observe mRAN expression of MIF and iNOS in LPS-stimulated RAW 264.7 cells, RT-PCR was used. NO production in LPS-stimulated RAW 264.7 cells was measured by nitrite assay. All the female BALB/c mice were bred and maintained in pathogen-free mouse colonies and were 6 weeks of age on beginning of the experiment. The experimental model of RA was induced by injection of $50{\mu}g/kg$ LPS. Ulmus davidiana Planch herbal acupuncture solution was injected into either S 35 (犢鼻) or EX-LE 202 (內膝眼) of mice in turn daily for 19 days. Immunohistochemical staining was carried out to assess $TNF-{\alpha}$, $NF-{\kappa}B$ p65 and iNOS expression in synovial membrane. Synovial hyperplasia, angiogenesis and fibrosis in synovial membrane was observed with a microscope. Results : 1. Ulmus davidiana Planch herbal acupuncture solution inhibited mRNA expression of MIF and iNOS in dependence on a density of it in LPS-stimulated RAW 264.7 cells. 2. Ulmus davidiana Planch herbal acupuncture solution decreased synovial hyperplasia, angiogenesis and fibrosis in LPS-stimulated mouse knee joints. 3. Ulmus davidiana Planch herbal acupuncture solution curtailed production of MIF, $TNF-{\alpha}$, $NF-{\kappa}B$ p65, iNOS in LPS-stimulated mouse knee joints. Conclusion : On the basis of these results, It was shown that Ulmus davidiana Planch herbal acupuncture solution is significantly able to inhibit the production of MIF as a top in cytokines related to inflammatory or irrlll1une responses. Our results may provide that Ulmus davidiana Planch herbal acupuncture solution has beneficial effect in not only RA but other inflammatory or immune deases.

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법랑아세포종에서 p21 및 p53 발현에 관한 면역조직화학적 연구 (IMMUNOHISTOCHEMICAL STUDY OF P21 AND P53 EXPRESSION IN AMELOBLASTOMA)

  • 신동준;명훈;황경균;김명진
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제29권4호
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    • pp.199-205
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    • 2003
  • The p53 protein was discovered in 1979 as cellular 53-kD nuclear phosphoprotein bound to the large transforming antigen of SV40 virus. $P21^{WAF1/CIP1}$, which has been described as the critical downstream mediator of p53, is known to suppress DNA replication and arrest the G1 cell cycle by quaternary complex with cyclin D, cyclin-dependent kinase(CDK) and proliferating cell nuclear antigen(PCNA). In these days, some studies shows that the p21 can be induced by independent pathways. There are various reports about the expression of p21 (67%.82.4%) in oral squamous cell carcinoma. But these studies are mostly done in malignant tumor not in benign tumor. So we decided to study the expression of p21 in ameloblastoma and the relationship between p53 and p21 as a downstream mediator of p53 in ameloblastoma. We investigated the expression of p21 and p53 with the method of immunohistochemistry. We selected 30 cases of ameloblastoma tissue blocks (acanthomatous type: 5 cases, follicular type: 8 cases, plexiform type: 17 cases) imbedded in paraffin. We used 30 cases of normal gingival tissues and 30 cases of squamous cell carcinoma tissues (SCC) respectively and compared their results with those of ameloblastoma. We made slides with the streptavidin-biotin methods and used monoclonal antibody DO-7 (Novocastra, Newcastle, United Kingdom) as p53 antibody and monoclonal antibody M7202 (DAKO, California, U.S.A.) as p21 antibody. We used Pearson's correlation coefficient to analyse the relationship. The results were as follows: 1. p21 was expressed in ameloblastoma about 30% and this is lower than that of normal gingiva and SCC. 2. In normal gingiva and ameloblastoma, p21 expression was correlated with p53 expression. 3. In SCC, p21 were expressed about 83.3% and this is more than that of p53. But there was no correlation between p21 and p53 expression. We confirmed p21 expression and relation with p53 in ameloblastoma. But, to confirm the function of p21, more studies about p21 expression in malignant ameloblastoma and ameloblastic carcinoma are needed.

하악골 신장술 후 신생골 조직에서 자가분비성장능력의 활성에 대한 실험적 연구 (AN EXPERIMENTAL STUDY FOR THE DETECTION OF AUTOCRINE GROWTH ACTIVITY IN THE OSTEOGENIC CELLS AFTER MANDIBULAR DISTRACTION OSTEOGENESIS)

  • 변준호;박봉욱;박성철;김규천;박봉수;김종렬
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제33권4호
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    • pp.331-339
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    • 2007
  • Background: Distraction osteogenesis(DO) is a useful method for treating cases demanding the generation of new bone. During DO, the angiogenic activity is crucial factor in the new bone formation. The aim of this study was to detect the autocrine growth activity in the cellular components of the distracted bone with observation of the co-expression of vascular endothelial growth factor(VEGF) and its receptors following the mandibular DO. Materials and methods: Unilateral mandibular distraction(0.5 mm twice per day for 10 days) was performed in six mongrel dogs. Two animals were killed at 7, 14, and 28 days after completion of distraction, respectively. Immediately after the animals were killed, the right mandibles were harvested en block. Immunohistochemical staining was processed for observation of the VEGF expression, and double immunofluorescent staining was also processed for detection of the co-expression of osteocalcin and VEGF's two distinct receptors(VEGFR-1 and VEGFR-2). Results: At 7 and 14 days after distraction, the expressions of VEGF were significantly increased in the osteogenic cells of the distracted bone. Up to 28 days after distraction, VEGF was still expressed moderate in the osteoblastic cells of distracted bone. The co-expressions of osteocalcin/VEGFR-1 and osteocalcin/VEGFR-2 were observed in the distracted bone at 7 and 14 days after distraction. In the double immunofluorescent staining, the co-expression' s level of osteocalcin/VEGFR-1 was more than that of osteocalcin/VEGFR-2. Conclusion: Taken together, this study suggested that VEGF plays an important role in the osteogenesis, and these osteoblastic cell-derived VEGF might act as autocrine growth factor during distraction osteogenesis. In the other word, the cellular components, such as osteoblasts and immature fibroblast-like cellsor mesenchymal cells in the distracted bone, might have autocrine growth activity during distraction osteogenesis.

위와 소장에 생긴 위장관 간질종양의 임상병리학적 비교 (Comparative Study of the Clinicopathologic Characteristics between Gastrointestinal Stromal Tumors Arising from the Stomach and Small Bowel)

  • 허건웅;신동우;백소야;김일동;김기호;서병선;박진수;김상욱
    • Journal of Gastric Cancer
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    • 제7권4호
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    • pp.254-260
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    • 2007
  • 목적: 위와 소장에 발생한 위장관 간질종양의 임상병리학적 특징을 비교하여 예후 인자 및 적절한 치료의 방침을 알아보고자 하였다. 대상 및 방법: 대진의료재단 분당제생병원에서 1998년 8월부터 2006년 5월까지 위장관 간질종양으로 진단된 38명의 환자 중 분석 및 추적 조사가 가능했던 29명의 환자에서 임상 양상을 조사하고, 면역조직화학적 염색을 시행하였으며, NIH 합의안에 따라 위험도를 분류하여 각각 위와 소장에서 발생한 위장관 간질종양을 비교하였다. 결과: 위와 소장에 발생한 위장관 간질종양의 임상병리학 적 차이 및 재발 양상의 차이는 없었으며, NIH 위험도 분류에 따라 나눈 고위험군과 저위험군 간에 재발의 차이는 있었다(P=0.030). 결론: 위장관 간질종양에서 원발 부위인 위와 소장간에 임상병리학적 양상이나 예후에 통계학적으로 유의한 차이는 없었으나, NIH 분류에 의한 고위험군에서는 재발 가능성이 높으므로 치료 지침에 따른 적절한 추적관찰이 필요하며, 앞으로 여러 기관의 예를 통합한 큰 모집단을 대상으로 지속적인 연구를 시행하여 국내 실정에 맞는 정확한 진단 기준 및 치료지침이 만들어져야 될 것으로 사료된다.

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아토피양(樣)피부염 NC/Nga 생쥐에서 자음제습탕가감(滋陰除濕湯加減)과 아토피 크림-자운고(紫雲膏)의 병용투여가 피부염에 미치는 영향 (Effects of Concurrent Administration of JaUmJeSeupTangKaKam(JUJSTK) and Atopy Cream, Jawoongo(AJ) on Atopic Dermatitis-like Skin Lesions in NC/Nga Mouse)

  • 이남열;김윤희;한재경
    • 대한한방소아과학회지
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    • 제23권3호
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    • pp.9-36
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    • 2009
  • Objectives The purpose of this study is to examine the effect of a concurrent administration of JUJSTK and AJ on atopic dermatitis in an in-vivo experiment. Thus, this study is expressed by using NC/Nga atopic dermatitis mice which have histological and clinical similarities to that of humans have been used. Methods Clinical skin score, hematology, serum total IgE and IgG1 of the mouse was evaluated, and cytokine levels, total number of the cells, immunohistochemical staining, histological features of axillary lymph node(ALN), peripheral blood mononuclear cells(PBMCs), and a dorsal skin tissue of the mouse were analyzed. Results Oral administration of JUJSTK and concurrent administration of JUJSTK and AJ lowered the clinical skin score, total cell number of WBC, eosinophils in blood, and serum total of IgE & IgG1, IFN-$\gamma$, IL-5, IL-13, IL-17. In addition, total cell number of ALN and dorsal skin tissue, absolute cell number of $CD3e^+$ T cell, $CD4^+$ Th cell, $CD8^+$ c/sT cell, $CD3^+CCR3^+$ cell, $CCR3^+$ cell, $CD3^+CD69^+$, $CD4^+CXCR5^+$ in ALN, PBMCs, absolute cell number of $CCR3^+$, $CD3^+/CD69^+$, $CD11b^+/Gr-1^+$, $CD11b^+/Gr-1^+$ in dorsal skin tissue, Eotaxin2 mRNA, CCR3 mRNA in dorsal skin tissue and gene expression of IL-5 mRNA, IL-13 mRNA in ALN were significantly decreased. Furthermore, thickness of epidermis infiltrated inflammatory immune cell & mast cell in dermis, histological infiltration of mast cell, the size of inflammatory lymphocytes cells & plasma cells in ALN and histological infiltration of $CD4^+$ & $CCR3^+$ in ALN and dorsal skin tissue were significantly decreased as well. Conclusions Concurrent administration of JUJSTK and AJ on atopic dermatitis in an in-vivoexperiment by using an NC/Nga atopic dermatitis mouse was very effective as an atopic dermatitis treatment.

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Expression of TRPV1 and iNOS in the Dorsal Root Ganglion Exposed by Autologous Nucleus Pulposus in the Rat

  • Kim, Su-Jeong;Seo, Jeong-Min;Cho, Yun-Woo;Park, Hea-Woon;Lee, Joon-Ha;Hwang, Se-Jin;Ahn, Sang-Ho
    • The Journal of Korean Physical Therapy
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    • 제22권3호
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    • pp.71-77
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    • 2010
  • Purpose: To determine whether upregulation of inducible nitric oxide synthase (iNOS) transcription and translation is related to radicular pain in a model of lumbar disc herniation. Also, to investigate the temporal changes of mRNA expression of iNOS and the identity of iNOS and transient receptor potential vanilloid (TRPV) 1 channel expression cells in dorsal root ganglion (DRG) of a model of lumbar disc herniation. Methods: A lumbar disc herniated rat model was developed by implantation of the autologous nucleus pulposus, harvested from the coccygeal vertebra of each tail, on the left L5 nerve root just proximal to the DRG. Rats were tested for mechanical allodynia of the plantar surface of both hind paws 2 days before surgery and 1, 5, 10, 20 and 30 days postoperatively. Reverse transcription polymerase chain reaction (RT-PCR) was used to follow iNOS mRNA expression. To stain iNOS and TRPV1 in DRG, an immunohistochemical study was done 10 days after surgery. Results: A significant drop in mechanical withdrawal threshold on the ipsilateral and contralateral hind paws was observed 1 day after surgery and was prolonged to 30 days in rats with lumbar disc herniation. The expression of mRNA for iNOS peaked at postoperative day 10 on both sides of the DRG. iNOS-positive sensory neurons in the DRG varied in size from large to small diameter cells. A majority of small and intermediate sensory neurons were TRPV1-positive cells. Double immunofluorescence staining for TRPV1 and iNOS revealed that most intermediate TRPV1-positive sensory neurons co-localized with iNOS-positive neurons. Conclusion: Nucleus pulposus-induced mechanical allodynia can be generated without mechanical compression. This pain is related to temporal changes in expression of iNOS mRNA in the DRG. Co-localization of TRPV1 and iNOS in intermediate neurons of the DRG is correlated with pain modality and intensity.

아프리카 왕달팽이(Achatina fulica) 뇌신경절의 면역조직화학적 구조 (Immunohistochemical Study on the Cerebral Ganglion of African giant Snail, Achatina fulica)

  • 장남섭;한종민
    • 한국패류학회지
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    • 제15권1호
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    • pp.1-11
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    • 1999
  • 아프리카 왕달팽이(Achatina fulica)의 뇌신경절을 AB/AY 염색법과 면역조직화학법을 이용, 실험한 결과 다음과 같은 결론을 얻었다. 아프리카 왕달팽이의 뇌신경절은 2 x 1 mm 정도 크기의 흰색 타원형체로 중앙은 직경 1 mm 정도의 대뇌교련부(cerebral commissure)에 의해 연결되어 있었다. 뇌신경절(cerebral ganglion)의 종단면은 나비모양으로, 세포의 분포형태에 따라 중배부위(medio-dorsal part), 측배부위(latero-dorsal part), 미배부위(caudo-dorsal part) 그리고 측엽부위(lateral lobe) 등으로 나눌 수 있었다. 중배부위와 측배부위를 구성하는 대부분의 세포는 LG 세포와 약간의 DG 세포로서, 이들은 혼재되어 나타났다. 그러나 측엽부위에서는 LG 세포와 DG 세포는 거의 관찰되지 않고, 주로 Y 세포만이 관찰되었다. LG 세포는 크기가 20-70 $\mu\textrm{m}$ 정도의 원형 또는 타원형의 세포로서 AB/AY 염색시 밝은 녹색으로 염색되고 핵질속에서는 과립상의 염색질이 고르게 분비되어 있으며, 1-3 개의 인이 관찰되었다. 세포질 속에는 분비성과립들이 고르게 발달해 있었다. DG 세포는 대부분 장타원형의 세포로서 LG 세포에 비해 약간 작았다. 이들은 m-b 이중염색에서 강한 methylenophilia를 보였고, AB/AY 염색에서는 진한 녹색으로 염색되었다. 이 세포는 rabbit anti-somatostatin antibody를 이용한 염색에서 노란색으로 염색되고, H-E 염색에서는 eosinophilia를 보여 호산성세포로 확인되었다. 그러나 BG 세포는 위의 세포들과는 달리 뇌신경절의 주변 부위 결합조직에서 관찰된 매우 드문 세포였다. 이들은 AB/AY 염색시 청녹색으로 염색되고, H-E 염색에서는 hematoxylin 에 강한 반응을 보였다.

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Central, Extraventricular and Atypical Neurocytomas: a Clinicopathologic Study of 35 Cases from Pakistan Plus a Detailed Review of the Published Literature

  • Ahmad, Zubair;Din, Nasir Ud;Memon, Aisha;Tariq, Mohammad Usman;Idrees, Romana;Hasan, Sheema
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권3호
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    • pp.1565-1570
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    • 2016
  • Background: Central neurocytomas are rare neuronal neoplasms with a favorable prognosis. They are typically located in the lateral ventricles of the brain and mostly histologically correspond to WHO grade II with a Mib 1 labelling index of <2%. Similar tumors located in the cerebral hemispheres and spinal cord, for example, are called "extraventricular neurocytomas". A few tumors histologically show atypia, mitoses, vascular proliferation and/or necrosis and a Mib 1 index >2 % and are designated as "atypical neurocytomas. Aim: The aim of our study was to describe the common as well as unusual morphologic features and the role of various immunohistochemical stains in the diagnosis of these rare tumors. Materials and Methods: We retrieved and reviewed 35 cases diagnosed between 2001 and 2015. Results: Sixty percent of patients were males, and the mean age was 26 years. 31 cases (88.6%) were intraventricular and 4(11.4%) were extraventricular. Histologically, 6 cases (17.1%) were compatible with "atypical neurocytomas". All cases showed the classic morphology comprising nests and sheets of uniform, round cells with uniform round to oval nuclei with finely speckled chromatin and perinuclear cytoplasmic clearing (halos). All cases also showed delicate, fibrillary, neuropil-like matrices. Other common histologic features included capillary-sized blood vessels in a branching pattern in 57.1%, foci of calcification in 34.3% and perivascular pseudorosettes in 20%. Rare findings included Homer-Wright or true rosettes in 8.6% and ganglioid cells in 2.9%. Synaptophysin was the most consistent and valuable marker, being positive in almost all cases. GFAP positivity in tumor cells was seen in 25.7% of cases. Follow up was available in 13 patients. Of these 9 had histologically typical and 4 had atypical tumors. Only 1 (with an atypical neurocytoma) died, probably due to complications of surgery within one month, while 12 (including 3 with atypical neurocytomas) remained alive. Recurrence developed in 1 of these 12 patients (histologically consistent with typical morphology) almost 9 years after surgery. Only 4 patients, including 2 with atypical tumors, received postoperative radiotherapy, all with surgery in 2010 or later. Overall, prognosis was excellent with prolonged, recurrence free survival and most patients, even without receiving radiation therapy, were alive and well for many years, even a decade or more after surgery, without developing any recurrence, indicating the benign nature of these neoplasms.

Expression of ER, PR, C-erbB-2 and Ki-67 in Endometrial Carcinoma and their Relationships with the Clinicopathological Features

  • Yu, Cui-Ge;Jiang, Xiang-Yang;Li, Bin;Gan, Lu;Huang, Jian-Feng
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권15호
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    • pp.6789-6794
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    • 2015
  • Background: To analyze the expression of estrogen receptors (ER), progesterone receptors (PR), C-erbB-2 and Ki-67 in endometrial carcinoma (EC) and their relationships with the clinicopathological features. Materials and Methods: Sixty-seven EC samples, 53 normal endometrial samples and 53 atypical hyperplasia endometrial samples were all selected in Shaanxi Provincial People's Hospital from Jun., 2012 to Jun., 2014. The expression of ER, PR, C-erbB-2 and Ki-67 in EC tissue, normal endometrial tissue and atypical hyperplasia endometrial tissue was respectively detected using immunohistochemical SP method. The relationships between the expression of ER, PR, C-erbB-2 and Ki-67 and the patients' clinicopathological features as well as their correlations in EC tissue were also analyzed. Results: The positive expression rates of ER and PR in EC tissue were 44.8% and 41.8%, respectively, dramatically lower than in atypical hyperplasia endometrial tissue and normal endometrial tissue (P<0.01). The positive expression rates of C-erbB-2 and Ki-67 in EC tissue were 80.6% and 64.2%, respectively, significantly higher than in atypical hyperplasia endometrial tissue and normal endometrial tissue (P<0.01). In EC tissue, the expression of ER and PR was closely associated with the differentiated degrees and depth of myometrial invasion (P<0.05), while that of C-erbB-2 and Ki-67 with the clinical staging, differentiated degrees, depth of myometrial invasion and presence or absence of lymph node metastasis (P<0.05). Spearman correlation analysis further displayed that the expression of ER was positively correlated with PR (r=0.393, P=0.001), but negatively with C-erbB-2 and Ki-67 (r=-0.469, P=0.000; r=-0.329, P=0.007); The expression of PR was negatively correlated with C-erbB-2 and Ki-67 (r=-0.273, P=0.025; r=-0.251, P=0.041), but that of C-erbB-2 positively with Ki-67 (r=0.342, P=0.005). Conclusions: Abnormal expression of ER, PR, C-erbB2 and Ki-67 might play an important role in endometrial malignant transformation and cell differentiation, so their joint detection is likely to be a comprehensive combination of immune factors, which is of great importance for EC prognosis.

Expression of Endogenous Hypoxia Markers in Vulvar Squamous Cell Carcinoma

  • Li, Yu-Zhu;Li, Shu-Ling;Li, Xia;Wang, Li-Jie;Wang, Jiu-Ling;Xu, Jia-Wen;Wu, Zhi-Hong;Gong, Li;Zhang, Xiao-Dan
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권8호
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    • pp.3675-3680
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    • 2012
  • Objective: To investigate the expression of endogenous hypoxia-related markers identified as being involved in vulvar squamous cell carcinoma (VSCC). Methods: We performed immunohistochemical staining of hypoxia-inducible factor-$1{\alpha}$ (HIF-$1{\alpha}$), glucose transporter-1 (GLUT-1), carbonic anhydrase 9 (CA-9) and vascular endothelial growth factor (VEGF), on tissue sections of 25 VSCC patients, 10 vulvar intraepithelial neoplasia (VIN) patients and 12 healthy controls. Results: HIF-$1{\alpha}$ expression was found in all sections, with no significant difference between controls, VIN and VSCC sections (all P<0.05). Glut-1 expression was found in 25% of control, 90% of VIN and 100% of VSCC sections. A significant difference between control and VIN or VSCC was observed (all P<0.05), while no difference was found between VIN and VSCC sections (P>0.05). CA-9 expression was negative in control sections, but it was found in 30% of VIN sections and 52% of VSCC sections with strong staining. Similarly, CA-9 expression also showed obvious differences between controls and VIN or VSCC sections (all P<0.05). However, there was no significant difference between VIN and VSCC (P>0.05). There were only 25% of control sections with weak VEGF expression, while strong staining was found in about 60% of VIN sections and 25% of VSCC sections (all P<0.05). In addition, a difference was also found between VIN and VSCC sections (P<0.05). Conclusion: Expression of endogenous hypoxia markers (HIF-$1{\alpha}$, GLUT-1, CA-9 and VEGF) might be involved in the malignant progression of VSCC.