Involvement of VEGF in Both Cell Apoptosis and Survival in the Retina of Type 2 Diabetic OLETF Rats

김영희(Young Hee Kim),최미영(Mee Young Choi),김윤숙(Yoon Sook Kim),노구섭(Gu Seob Roh),김현준(Hyun Joon Kim),강상수(Sang Soo Kang),최완성(Wan Sung Choi),조경제(Gyeong Jae Cho) 대한해부학회 2007 Anatomy & Cell Biology Vol.40 No.4

Vascular endothelial growth factor (VEGF)는 망막혈관장애, pericyte 손실, 망막신경세포사와 세포증식등을 포함한 당뇨병의 초기망막병변에 많은 영향을 미친다. 본 연구에서는 제2형 당뇨병 쥐의 망막에서의 세포사와 세포생존에 VEGF가 연계되어 있는지를 조사하였다. 실험에는 자발적인 제2형 당뇨병 모델의 하나인 28주령의 Otsuka Long-Evans Tokushima Fatty (OLETF)쥐들과 대조군으로 Long-Evans Tokushima Otsuka (LETO)쥐들의 망막이 이용되었다. 대조군에 비해 OLETF 쥐의 망막에서는 pericyte 손실의 증거와 함께 세포증식, 세포사, endothelial nitric oxide synthase (eNOS)와 VEGF의 증가가 확인되었다. 또한, OLETF 쥐의 망막에서의 세포사멸신호는 VEGF에 특이적인 세포에서 일부 확인되었지만, VEGF-independent eNOS 특이 세포에서는 전혀 관찰되지 않았다. 이 결과들은 제2형 당뇨병 OLETF 쥐의 망막에서 VEGF는 세포사와 eNOS 의존적 세포생존에 모두 관계하고 있음을 시사한다. Vascular endothelial growth factor (VEGF) is closely involved in early retinal pathology of diabetes, including blood-retinal barrier breakdown, pericyte loss, neuro-retinal apoptosis, and cell proliferation. This study examines the involvement of VEGF in cell apoptosis and survival in the retina of animals with type 2 diabetes. We used retinas from 28-week-old Otsuka Long-Evans Tokushima Fatty (OLETF) rats, a model of spontaneous type 2 diabetes, and Long-Evans Tokushima Otsuka (LETO) rats as controls. In parallel with evidence for pericyte loss, we found cell proliferation, apoptosis, and endothelial nitric oxide synthase (eNOS) (an indicator of endothelial cell proliferation/survival) and VEGF overexpression in the OLETF-retina, compared to control LETO. Furthermore, apoptotic signals were partly co-localized to only VEGF-positive cells in the OLETF-retina, but no apoptotic signals were found in VEGF- and eNOS-double-positive cells. These results suggest that upregulated VEGF is involved in apoptosis and eNOS-dependent cell survival in the retinas of type 2 diabetic rats.

구강 편평세포암종에서 MMP-2, MMP-9 발현과 VEGF와 연관성

최수임,조의성,윤혜경,이희철 대한구강악안면병리학회 2003 대한구강악안면병리학회지 Vol.27 No.2

MMPs are catalytic enzymes involved in the degradation of extracellular marix, and associated with invasive growth and metastasis of malignant tumors along with angiogenesis. This study was to evaluate the prognostic significance of VEGF expression and microvessel density(MVD) and the relationship between VEGF expression, MVD and MMPs expression in oral squamous cell carcinomas(OSCC). The materials were from 52 cases of OSCC during a period from 1991 to 2001. Clinicopathologic parameters such as clinical stage, recurrence, histologic grade and invasion pattern were evaluated. Immunohistochemical staining for MMP-2, MMP-9, VEGF and CD34 were performed and statistical analyses between clinicopathologic parameters and VEGF expression and MVD were done. MMP-2, MMP-9 and VEGF expressions were noted in 30(57.7%), 21(40.1%) and 38(73.1%) of 52 cases, respectively. MVD was measurable in 35 cases, and cases with increased MVD more than average were 16(45.7%) of 35 cases. There was no significant relationship between clinicopathologic parameters and VEGF expression or MVD in OSCC, which suggests that VEGF expression and MVD can not be regarded as reliable prognostic factors. Cases with less infiltrative growth pattern showed a tendency of increased MVD, which can explain the possibility that neovascularization might be an early event of tumor invasion. Lack of significant relationship between MMPs, VEGF expressions and MVD might be due to limited number of cases, and positive correlation between MMP-2 and VEGF expression suggests that both factors might be involved in the process of angiogenesis in OSCC.

Expression of vascular endothelial growth factor mRNA and protein and vascular endothelial growth factor receptors in eutopic endometrium of patients with advanced endometriosis

( Sung Eun Hur ),( Ji Young Lee ),( Hye Sung Moon ),( Hye Won Chung ) 대한산부인과학회 2008 Journal of Womens Medicine Vol.1 No.1

Objective: To investigate the expression of vascular endothelial growth factors (VEGFs) and VEGF receptor 1 and 2 in eutopic endometrium of patients with advanced endometriosis and to understand the role of VEGFs and VEGF receptors in the development of endometriosis. Methods: Endometrial samples were obtained from 65 premenopausal women, 29~44 years of age, undergoing laparoscopic surgery or hysterectomy for non-malignant lesions. Sufficient samples were collected from 33 patients with endometriosis (stages III or IV) and 32 controls without endometriosis, as confirmed by laparoscopic surgery. Expression of VEGFs mRNA and protein and their receptors in the eutopic endometrium were analyzed by RT-QC PCR and Western blotting. The quantitative expression of VEGF-A, -B, and -C in eutopic endometrium from patients with endometriosis was examined throughout the menstrual cycle, and was compared to eutopic endometrial expression in control patients without endometriosis. Results: VEGF-A expression in healthy controls was lower in the secretory phase than in the proliferative phase. In patients with endometriosis, VEGF-A expression was not lower in the secretory phase than in the proliferative phase and was higher than in the secretory phase of healthy controls. The expression of VEGF-B and -C was similar to VEGF-A. The mRNA of VEGF receptors 1 and 2 was expressed at the same level in eutopic endometrium from patients with endometriosis and in healthy controls throughout the menstrual cycle. All eutopic endometrial samples from women with and without endometriosis in both the proliferative and secretory phases of the menstrual cycle showed clear bands at the expected molecular weights for VEGF-A and -B. Conclusions: These results suggest that specific angiogenic factors (VEGF-A, -B, and -C) play important roles in the pathogenesis of endometriosis.

구강점막 편평상피세포암에서 림프관형성 유전자 발현

박영욱,김성곤,김소희,김한석,김민근,Park, Young-Wook,Kim, Seong-Gon,Kim, So-Hee,Kim, Han-Seok,Kim, Min-Keun 대한악안면성형재건외과학회 2009 Maxillofacial Plastic Reconstructive Surgery Vol.31 No.6

Background and Purpose: Vascular endothelial growth factor (VEGF)-C, VEGF-D and their tyrosine kinase receptor, VEGF receptor (VEGFR)-3 are recently known to have lymphangiogenic activities in various tumor types. Oral mucosal squamous cell carcinoma (OMSCC) easily metastasizes to cervical lymph nodes, so we determined the expression levels of VEGF-C, VEGF-D and VEGFR-3 in oral squamous cell carcinoma. Materials and Methods: We performed Western blot analyses with 4 OMSCC cultured tumor cell lines (SCC9, KB, YD-10B, YD-38), and with 7 surgical specimens of OMSCC for the detection of VEGF-C, VEGF-D and VEGFR-3 proteins. Expression of VEGF-C mRNA as well as mRNA for VEGFR-3 in 4 OMSCC cell lines (KB, SCC-4, SCC-9, YD-10B) was investigated by RT-PCR. We also measured VEGFC/VEGF-D protein concentrations in the media and protein concentration of VEGFR-3 in cell lysates of 4 OMSCC cell lines (SCC9, KB, YD-10B, YD-38) using commerical ELISA kits. Finally, we performed immunoprecipitation for the detection of VEGF-C in cell lysates of 4 OMSCC cells (KB, SCC-4, SCC-9, YD-10B) and real-time RT-PCR for the quantification of VEGF-C mRNA. Results: In the result of Western blotting with cell lysates of 4 OMSCC cells, we could not detect the protein expression of VEGF-C, VEGF-D, and VEGFR-3. But, all tumor tissues demonstrated VEGF-C and VEGFR-3. VEGF-C mRNA was detected at various levels in 4 OMSCC cell lines. Moreover, OMSCC cells secreted VEGF-C, not VEGF-D and VEGFR-3 was also detected in cell lysates of OMSCC by ELISA. Immunoprecipitation and real-time RT-PCR revealed VEGF-C was also expressed in 4 OMSCC cell lines. Conclusion: Taken together, tumor cells of OMSCC secrete VEGF-C, not VEGF-D. And VEGFR-3 is expressed tumor cells as well as OMSCC tumor tissues, needs further study.

Multi-paratopic VEGF decoy receptor have superior anti-tumor effects through anti-EGFRs and targeted anti-angiogenic activities

Lee, Dae Hee,Lee, Myeong Youl,Seo, Youngsuk,Hong, Hyo Jeong,An, Hyun Joo,Kang, Jong Soon,Kim, Ho Min Elsevier 2018 Biomaterials Vol.171 No.-

<P><B>Abstract</B></P> <P>Limitation of current anti-Vascular Endothelial Growth Factor (VEGF) cancer therapy is transitory responses, inevitable relapses and its insufficient tumor-targeting. Thus, multifaceted approaches, including the development of bispecific antibodies and combination strategies targeting different pathways have been proposed as an alternative. Here, we developed a novel multi-paratopic VEGF decoy receptor, Cetuximab-VEGF-Grab and Trastuzumab-VEGF-Grab, by genetically fusing VEGF decoy receptor (VEGF-Grab) to a single chain Fv of anti-Epidermal Growth Factor Receptor (EGFR) antibody (Cetuximab and Trastuzumab). These multi-paratopic VEGF decoy receptor, which recognize VEGF and EGFR family (EGFR or HER2), effectively suppressed both VEGF and EGFR pathways <I>in vitro</I>, to levels similar to those of the parental VEGF-Grab and anti-EGFR antibodies. In addition, the concurrent binding of multi-paratopic VEGF decoy receptor to VEGF and EGFR family enabled their specific localization to EGFR + tumor <I>in vitro</I> and <I>in vivo.</I> Furthermore, Cetuximab-VEGF-Grab and Trastuzumab-VEGF-Grab exhibited the enhanced anti-tumor activities compared to VEGF-Grab in EGFR + tumor xenograft mouse model via anti-EGFR and the targeted anti-angiogenic activities. These results indicate that multi-paratopic VEGF decoy receptor can be a promising agent, combining tumor-targeted anti-angiogenic therapy with efficient blockade of proliferative signals mediated by EGFR family.</P>

VEGF-C and VEGF-D Expression and its Correlation with Lymph Node Metastasis in Esophageal Squamous Cell Cancer Tissue

Yang, Zeng,Wang, Yong-Gang,Su, Kai Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.1

Background: To explore vascular endothelial growth factor C (VEGF-C) and VEGF-D expression and its correlation with lymph node metastasis in esophageal squamous cell cancer (ESCC) tissue. Materials and Methods: Immunohistochemical methods were applied to detect the levels of VEGF-C and VEGF-D expression in 64 surgicall removal ESCC tissues, tissues adjacent to cancer and normal tissues, and the relationship between VEGF-C and VEGF-D expression and lymph node metastasis was analyzed. Results: Both VEGF-C and VEGF-D were expressed by varying degrees in esophageal cancer tissue, the tissue adjacent to cancer and normal tissue, and the positive expression rate went down successively. The positive expression rates of VEGF-C (59.4%) and VEGF-D (43.8%) in esophageal cancer tissue were significantly higher than in the tissue adjacent to cancer (34.4%, 15.6%) and normal tissue (20.3%, 12.5%), respectively, in which significant differences were manifested (p<0.01). Positive expression rates of VEGF-C and VEGF-D in esophageal cancers with lymph node metastasis were markedly higher than without such metastasis (p<0.01), while those in the tissue with TNM staging I~II were markedly lower than that with TNM staging III~IV (p<0.01). Conclusions: Both VEGF-C and VEGF-D are highly expressed in ESCC tissue, which may be related to the lymph node metastasis of cancer cells. Hence, VEGF-C and VEGF-D can be clinically considered as important reference indexes of lymph node metastasis in esophageal cancer.

임신한 흰쥐에서 자궁동맥 결찰에 의한 혈관내피성장인자의 변동

김용욱(Yong Wook Kim),이종건(Jong Kun Lee),이재성(Jae Sung Lee),최옥춘(Ok Choon Choi),노덕영(Duck Yeong Ro),김태응(Tae Eung Kim),정재근(Jae Geun Jung),신종철(Jong Chul Shin) 대한산부인과학회 2001 Obstetrics & Gynecology Science Vol.44 No.12

N/A Objective : During pregnancy, the impaired placental perfusion causes complications such as preeclampsia, intrauterine growth restriction and fetal death in utero. In order to investigate the maternal and fetal response to the impaired placental perfusion, the author induced the impaired placental perfusion by the ligation of the rat uterine artery and investigated its effect on the expression of VEGF (vascular endothelial growth factor) in the placenta and serum VEGF level. Methods : The rats on day 15 of gestation were used for the experiment. They were divided into two groups. The control group consists of the 20 rats that underwent laparotomy without uterine artery ligation. The experimental group consists of the 20 rats that underwent laparotomy and the uterine artery ligation by silk on day 15 of gestation. On day 16, 17, 18 and 19 of gestation, the placental tissues were obtained. The Mrna expressions of the VEGF in the placenta were measured by the relative RT-PCR in the control and experimental group. The localization and intensity of immunohistochemical staining of VEGF in placenta were determined in both groups and the maternal serum levels of VEGF were also measured in both groups. Results : The Mrna expressions of VEGF120 and VEGF164 were significantly increased 48 hours after the ligation (day 17 of gestation) but the Mrna expression of VEGF188 was not changed after the ligation. There was no difference in the location and intensity of immunohistochemical staining of VEGF in the placenta between control and experimental groups. The serum VEGF levels of control group were 9 times as high as those of non-pregnant rats. The significant increases of the serum VEGF levels were noted 48 and 72 hours after the ligation (day 17 and 18 of gestation) but the significant increase was not noted 96 hours after the ligation (day 19 of gestation) as compared to control group. Conclusion : This study demonstrated firstly that the experimentally induced reduction of placental perfusion increased expressions of VEGF in the placenta and maternal serum. The results support that the measurement of maternal serum VEGF levels in pregnancy may help the diagnosis of placental insufficiency.

성상세포종양에서 VEGF발현과 혈관신생 및 세포자멸사와의 상관성

김두천,이민철,김세종,김두천,이민철,김세종 대한신경과학회 1999 대한신경과학회지 Vol.17 No.3

수술로 절제된 비소세포폐암 조직에서 예후인자로서 VEGF와 bFGF 발현의 의의

김승준 ( Seung Joon Kim ),이정미 ( Jung Mi Lee ),김진숙 ( Jin Sook Kim ),강지영 ( Ji Young Kang ),이상학 ( Sang Hak Lee ),김석찬 ( Seok Chan Kim ),이숙영 ( Sook Young Lee ),김치홍 ( Chi Hong Kim ),안중현 ( Joong Hyun Ahn ),권순석 대한결핵 및 호흡기학회 2008 Tuberculosis and Respiratory Diseases Vol.64 No.3

연구배경: 혈관신생은 종양의 성장과 유지 및 전이에 필수적이며 따라서 종양조직은 혈관신생을 위해 많은 종류의 혈관형성 촉진인자들을 생성하고 있다. VEGF와 bFGF는 혈관신생과 관련되는 물질로 본 연구에서는 폐암환자에서 조직 내 VEGF와 bFGF의 발현에 대해 알아보고자 하였다. 방법: 조직학적으로 샘암종 또는 편평상피세포암종으로 진단받고 완치의 목적으로 수술을 시행 받은 35명의 폐암환자 조직에서 VEGF 및 bFGF의 농도를 ELISA 방법으로 측정하였으며 이에 대한 임상적 양상을 후향적으로 분석하였다. 결과: VEGF 및 bFGF의 농도는 종양조직이 대조조직보다 유의하게 높았으며 T2+T3의 종양조직이 T1의 종양 조직보다 유의하게 VEGF의 농도가 높았다. 림프절 전이가 있었던 경우가 없었던 경우보다 종양조직에서 VEGF의 농도가 증가한 경향을 보였다(p=0.06). 하지만 VEGF 및 bFGF의 농도는 환자의 병기 및 생존율에 통계적으로 유의한 차이를 보이지 않았다. 결론: VEGF 및 bFGF 모두 종양조직에서 증가하였으나 VEGF만이 종양크기, 림프절 전이 등의 임상양상과 연관성을 보여주었다. 하지만 각각의 VEGF 및 bFGF의 종양조직 내 농도는 예후와 관련되지는 않았다. Background: Tumor angiogenesis plays an important role in tumor growth, maintenance and metastatic potential. Tumor tissue produces many types of angiogenic growth factors. Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) have both been implicated to have roles in tumor angiogenesis. In this study, the expression of tissue VEGF and bFGF from non-small cell lung cancer (NSCLC) patients were analyzed. Methods: We retrospectively investigated 35 patients with a histologically confirmed adenocarcinoma or squamous cell carcinoma of the lung, where the primary curative approach was surgery. An ELISA was employed to determine the expression of VEGF and bFGF in extracts prepared from 35 frozen tissue samples taken from the cancer patients. Results: VEGF and bFGF concentrations were significantly increased in lung cancer tissue as compared with control (non-cancerous) tissue. The VEGF concentration was significantly increased in T2 and T3 cancers as compared with T1 cancer. Expression of VEGF was increased in node-positive lung cancer tissue as compared with node-negative lung cancer tissue (p=0.06). VEGF and bFGF expression were not directly related to the stage of lung cancer and patient survival. Conclusion: Expression of VEGF and bFGF were increased in lung cancer tissue, and the expression of VEGF concentration in lung cancer tissue was more likely related with tumor size and the presence of a lymph node metastasis than the expression of bFGF. However, in this study, expression of both VEGF and bFGF in tissue were not associated with patient prognosis. (Tuberc Respir Dis 2008;64:200-205)

직장 결장암의 간전이 환자에서 VEGF-A, C, D 발현의 비교연구

전광식(Kwang-Sik Chun),이경하(Kyung-Ha Lee),송인상(In-Sang Song),김지연(Ji-Yeon Kim),김제룡(Je-Ryong Kim),안문상(Moon-Sang Ahn),이상일(Sang-Il Lee),박종현(Jong-Hyun Park),최송이(Song-E Choi),강대영(Dae-Young Kang),송규상(Kyu-Sang Son 대한외과학회 2009 Annals of Surgical Treatment and Research(ASRT) Vol.76 No.5

Purpose: We aimed to investigate the correlations between expressions of angiogenic cytokines VEGF-A, C, D of primary colorectal cancer and liver metastasis. Methods: We examined paraffin-embedded primary colorectal cancer tissue from 45 patients who had liver resection due to colorectal liver metastasis (metastasis group) and 37 patients who had surgical resection due to colorectal cancer only (control group). In the control group, local recurrence and distant metastasis had not occurred. Immunohistochemical staining for VEGF-A, C and D was performed. We analysed the correlations between expression of VEGF-A, C and D in primary colorectal cancer tissues and clinicopathologic parameters. Results: VEGF-A expressions of primary colorectal carcinoma were not different between the two groups. VEGF-C was more frequently expressed in the metastasis group (P=0.008) but VEGF-D was more expressed in the control group (P=0.003). Patients with VEGF-C negative and VEGF-D positive expression were predominant in the control group (P=0.020). Tumor location, T stage, lymph node metastasis and tumor differentiation were not related with the expressions of VEGF-A, C, D but only preoperative CEA was positively correlated with VEGF-A and C expression. Conclusion: Expressions of VEGF-C in primary tumor were more frequent in metastatic colorectal cancer and expressions of VEGF-D were more frequent in nonmetastatic colorectal cancer. More large-scale prospective studies for VEGF-C and D expression in colorectal cancer are necessary.