Tyrosine kinase A receptor (trkA): a potential marker in epithelial ovarian cancer

Objectives

To evaluate the role of trkA receptor as a potential tumor marker in serous epithelial ovarian cancer and its relationship with the angiogenic factors expression as vascular endothelial growth factor (VEGF) and nerve growth factor (NGF). Additionally, to examine whether NGF and VEGF secreted by epithelial ovarian cancer (EOC) explants and from epithelial ovarian cancer cell line (A2780) are involved in the process of angiogenesis, such as cellular proliferation, migration and differentiation of the human endothelial cell line (EA.hy926).

Methods

The mRNA levels of VEGF, NGF and trkA receptors were measured using PCR in 60 ovarian samples. Cellular localization and semi-quantitative estimation of VEGF, NGF, total trkA and p-trkA was performed using IHC in epithelial cells. NGF, total trkA and p-trkA protein were also evaluated in endothelial cells from the same tissues. Human endothelial cell line EA.hy926 was cultured with conditioned media obtained from both EOC explants and from the A2780 cell line, with or without NGF stimulus.

Results

Significantly higher levels of NGF, total trkA and p-trkA protein expressions were observed in epithelial and endothelial cells in poorly differentiated EOC versus normal ovary. Interestingly, the p-trkA receptor expression level showed the most significant difference and its presence was only found in borderline tumor and EOC samples indicating the importance of trkA receptor in EOC as a potential tumor marker.A significant increase in proliferation, migration and differentiation of EA.hy926 cells was observed with NGF, and this effect was significantly reverted when NGF was immuno-blocked and when a trkA inhibitor was used, showing that NGF is an important angiogenic factor in EOC by activating its trkA receptor.

Conclusion

These results indicate that p-trkA may be considered as a new potential tumor marker in EOC, and that NGF may also act as a direct angiogenic factor in EOC.     

血管内皮生长因子及其受体在卵巢上皮性癌组织中的表达

目的研究血管内皮生长因子(VEGF)及其受体(FLT1、FLK1)mRNA在卵巢上皮性癌组织中的表达及与临床病理因素的相关性。方法采用逆转录聚合酶链反应技术检测70例卵巢上皮性癌及22例卵巢良性肿瘤病变组织标本中VEGFmRNA及其受体FLT1mRNA及FLK1mRNA的表达。结果在良、恶性卵巢组织中均检测到VEGF121mRNA、165mRNA的表达,在卵巢上皮性癌组织中表达水平分别为(0.452±0.134,0.301±0.096)高于良性卵巢肿瘤[0.195±0.066(P=0.000),0.204±0.059(P=0.001)];在卵巢上皮性癌组织中,VEGF121mRNA表达高于VEGF165mRNA(P=0.000),而在良性卵巢肿瘤组织中两者表达水平差异无显著性(P=0.667)。FLT1mRNA、FLK1mRNA在部分卵巢上皮性癌组织中表达,分别为38.6%(27/70)和25.7%(18/70),而在良性卵巢肿瘤组织中未见表达。卵巢癌组织中VEGF121、VEGF165、FLT1、FLK1之mRNA表达与患者年龄、肿瘤体积、淋巴结转移、肿瘤分期之间无明显相关性。结论VEGF121、165及其受体FLT1、FLK1在卵巢上皮性癌的血管生成中起重要作用。     

肝细胞生长因子对卵巢癌血管生成因子的影响及信号传导途径

目的研究肝细胞生长因子(HGF)对人卵巢癌SKOV3细胞血管内皮生长因子的影响及其信号传导机制。方法将不同浓度、不同作用时间的HGF和核转录因子(NFkB)抑制剂PDTC刺激培养的SKOV3细胞;应用逆转录-聚合酶链反应技术(RT-PCR)检测卵巢癌细胞血管内皮生长因子(VEGF)mRNA的变化;采用蛋白印迹方法检测卵巢癌细胞VEGF蛋白和核蛋白NFkB的变化。结果HGF促进卵巢癌细胞VEGF mRNA和蛋白的表达,且随时间和浓度增加作用增强,PDTC可以抑制HGF的刺激作用;HGF促进细胞核NFkB蛋白的活化,且随时间延长作用增强,于刺激1 h达高峰,PDTC可以抑制HGF对NFkB的活化作用。结论HGF通过NFkB途径在核酸和蛋白水平调节卵巢癌细胞分泌VEGF。     

卵巢上皮性癌组织中肿瘤浸润性树突状细胞的状态及其与血管内皮生长因子表达的相关性

目的研究卵巢上皮性癌组织中肿瘤浸润性树突状细胞(TIDC)的状态及其与血管内皮生长因子(VEGF)表达的关系。方法采用免疫组化链霉菌抗生物素蛋白-过氧化酶连接法和Picture二步法,检测57例卵巢上皮性癌(恶性组)、30例良性卵巢上皮性肿瘤(良性组)及16例正常卵巢(正常组)组织中S-100蛋白、CD83标记的TIDC状态及其VEGF的表达。结果(1)恶性组TIDC光镜下形态基本可分成两种,其分布有异质性。恶性组S-100^ TIDC中位数为4．3个／400高倍视野(HPF),分别与良性组(中位数为1．8个／HPF)和正常组(中位数为2．0个／HPF)比较,差异有显著性(P值分别为0．000和0．015)。恶性组中,早期(Ⅰ～Ⅱ期)肿瘤组织中S-100^ TIDC数量明显多于晚期(Ⅲ～Ⅳ期,中位数分别为6．0个和3．8个／HPF,P=0．026)。恶性组中,CD83^ TIDC浸润肿瘤间质者极少,中位数为0个／HPF。(2)恶性组细胞中VEGF高表达(5．0分),分别与良性组(3．8分)和正常组(2,4分)比较,差异均有极显著性(P=0.000)。(3)恶性组中,S-100^ TIDC数量与VEGF表达呈明显负相关(P=0．001)。结论(1)卵巢上皮性癌细胞可刺激TIDC的募集,但同时受VEGF的抑制。(2)卵巢上皮性癌中TIDC成熟严重受抑。     

血管内皮生长因子mRNA异构体表达及与葡萄胎恶变的关系

目的 研究妊娠滋养细胞疾病（gestational trophoblastic disease，GTD）血管内皮生长因子（vascular endothelial growth factor，VEGF）mRNA异构体表达特征及其与葡萄胎恶变的关系。方法 采用逆转录-聚合酶链反应（RT—PCR）法对54例GTD组织和10例正常早孕绒毛组织VEGFmRNA异构体进行相对定量检测。结果 GTD组织和正常早孕绒毛组织表达VEGF121、VEGF145,VEGF165、VEGF189 mRNA，VEGF121表达水平显著高于其他3种异构体（P〈0．05）。4种异构体在GTD组织的表达水平显著高于正常早孕绒毛组织（P〈0．05）。恶变组葡萄胎VEGFmRNA表达量显著高于非恶变组葡萄胎（P〈0．01）。结论 GTD组织表达VEGF121,VEGF145、VEGF165、VEGF189mRNA，以VEGF121为主。恶变组葡萄胎VEGF mRNA表达量显著高于非恶变组葡萄胎。可能与葡萄胎恶变有关。     

EGFR、VEGF及LRP mRNA在卵巢上皮性癌组织中的表达及其意义

目的:探讨表皮生长因子受体(epidermal growth factor receptor,EGFR)、血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)和肺耐药蛋白(lung resistanceprotein,LRP)mRNA在卵巢癌的表达及意义。方法:用逆转录聚合酶链反应(RT-PCR)技术检测15例正常卵巢、13例卵巢良性肿瘤及51例卵巢上皮癌组织中EGFR mRNA、VEGF mRNA和LRP mRNA的表达,并分析它们的相关性。结果:卵巢上皮癌组织中EG-FR、VEGF和LRP阳性表达率显著高于正常卵巢及卵巢良性肿瘤组织(P<0.05);EGFRmRNA表达与卵巢上皮癌手术病理分期有关,Ⅲ~Ⅳ期的阳性表达率及表达强度高于Ⅰ~Ⅱ期(P<0.05);VEGF mRNA表达与卵巢上皮癌手术病理分期及淋巴结转移有关,Ⅲ~Ⅳ期和有淋巴结转移组的表达强度分别高于Ⅰ~Ⅱ期和无淋巴结转移组(P<0.05);LRP mRNA表达与卵巢上皮癌患者年龄、临床分期、分化程度、病理类型及淋巴结转移等临床病理参数无关(P>0.05);EGFR和VEGF基因之间(r=0.460,P<0.05)、EGFR和LRP基因之间表达显著相关(r=0.749,P<0.01)。结论:EGFR与卵巢癌血管生成和化疗耐药产生有关,可能参与了卵巢癌的发生发展,检测卵巢癌EGFR、VEGF和LRP可能对靶向化疗有指导作用。     

A-kinase anchoring protein 3 messenger RNA expression correlates with poor prognosis in epithelial ovarian cancer

OBJECTIVES: Cancer-testis (CT) antigens are expressed in tumors but not normal tissues except the testis and could be targets for vaccine therapy in epithelial ovarian cancer (EOC). A-kinase anchoring protein 3 (AKAP-3) is a novel member of the CT antigen family. The aim of this study was to examine the expression of AKAP-3 in EOC and correlate with clinico-pathologic characteristics. METHODS: One step RT-PCR was performed with RNA from normal and ovarian cancer cell lines and 74 epithelial ovarian tumor tissues. AKAP-3-specific PCR product was amplified. The distribution of AKAP-3 expression and clinico-pathologic variables was analyzed. Survival distributions were estimated, and multivariate analyses were performed. RESULTS: AKAP-3 mRNA expression was demonstrated in 43/74 (58%) of the ovarian cancer specimens. AKAP-3 was expressed in normal testis, but not in other normal tissues. AKAP-3 expression significantly correlated with increased likelihood of residual tumor (P = 0.005), but no increase in the likelihood of recurrence or persistent disease (P = 0.06). Patients with AKAP-3 mRNA expression were found to have a significantly poorer overall survival (median 50 months) compared with patients without AKAP-3 expression (median not reached) (P = 0.007). Multivariate analysis of AKAP-3 expression, residual disease, and response to frontline chemotherapy found response to be the strongest predictor of overall survival (P = 0.012). CONCLUSIONS: Our data demonstrate that AKAP-3 is expressed at high frequency in patients with EOC. Since AKAP-3 demonstrates tumor-restricted expression and appears to be associated with worse overall survival, it could represent an attractive target for antigen-specific immunotherapy in EOC.     

血管内皮生长因子与血小板源性生长因子在卵巢上皮性癌淋巴管形成中的作用

目的 探讨血管内皮生长因子(VEGF)和血小板源性生长因子(PDGF)在卵巢上皮性癌(卵巢癌)淋巴管形成中的作用.方法 RT-PCR技术检测淋巴管内皮细胞核标志物Proxl和淋巴管形成相关因子VEGF-A、VEGF-C、VEGF-D及PDGF-A、PDGF-B、PDGF-C、PDGF-D在卵巢癌细胞株SKOV3、70例卵巢上皮性肿瘤(卵巢良性肿瘤15例、卵巢交界性肿瘤10例、卵巢癌45例)和20例正常卵巢组织中的表达情况.实时定量PCR技术检测卜述90例卵巢组织中Proxl、VEGF-A、-C、-D及PDGF-A、-B、-C、-D的表达水平,并进行相关性分析.结果 (1)Proxl在各种卵巢组织中均有表达,而在SKOV3细胞中无表达;VEGF-A、-C、-D及PDGF-A、-B、-C、-D在SKOV3细胞和各种卵巢组织中均有表达.(2)卵巢癌组织中Proxl(2.2±1.3)、VEGF.A(3.5±1.5)、VEGF-C(19 ±14)、VEGF-D(3.0±1.8)及PDGF-A(3.3±3.3)、PDGF-C(6.9±4.6)的表达水平高于卵巢良性肿瘤和交界性肿瘤(P均＜0.05).(3)Proxl、VEGF-A和PDGF-A在卵巢癌Ⅲ～Ⅳ期(Proxl:2.6±1.3,VEGF-A:4.0± 1.4.PDGF-A:4.1±3.7)、淋巴结转移阳性(Proxl:3.0±1.4,VEGF-A:4.1±1.7,PDGF-A:4.9±4.1)及腹膜转移阳性(Proxl:2.8±0.9,VEGF-A:4.0±1.8,PDGF-A:4.5±4.0)的组织中的表达水平,分别高于Ⅰ～Ⅱ期、淋巴结转移阴性和腹膜转移阴性者(P均＜0.05);VEGF-C、VEGF-D在淋巴结转移阳性卵巢癌组织中的表达水平(VEGF-c:24± 13,VEGF-D:3.9±2.0)高于淋巴结转移阴性者(P均＜0.05).(4)卵巢癌组织中Proxl的表达水平与VEGF-D(r=0.62,P＜0.01)、PDGF-C(r=0.91,P＜0.01)、PDGF-D(r=0.61,P＜0.01)的表达水平呈正相关关系.结论 VEGF-A、VEGF-C和PDGF-A可能通过参与淋巴管形成之外的机制促进卵巢癌的淋巴结转移;VEGF-D可以促进卵巢癌的淋巴管形成及淋巴结转移;PDGF-B与卵巢癌的淋巴管形成及淋巴结转移无关;PDGF-C、PDGF-D参与卵巢癌淋巴管形成,但无促进淋巴结转移的作用.     

Absence of estrogen receptor-beta expression in metastatic ovarian cancer

OBJECTIVE: To evaluate the expression of estrogen receptor (ER)alpha and ERbeta mRNA and protein in normal ovarian tissue and primary and metastatic tumors. METHODS: Estrogen receptor alpha and ERbeta expression was studied in normal ovarian biopsies (n = 9) and primary (n = 8) and metastatic ovarian epithelial cancers (n = 8). Ovarian tissue was collected from surgical samples. Estrogen receptor alpha and ERbeta mRNA expression was compared by coamplification of the mRNA of the ERs. Expression was confirmed at the protein level by Western blot analysis using antibodies specific for each receptor. RESULTS: Among eight primary ovarian cancer samples, three had only ERalpha, two had only ERbeta, and three had both. All eight metastatic ovarian cancer tissues expressed only ERalpha mRNA and protein. Biopsies from normal ovaries had ERalpha and ERbeta mRNA and protein. Two of the ovarian epithelial cancer samples were paired and showed the same results. CONCLUSION: We found varying amounts of ERalpha and ERbeta in normal ovaries, lower levels of ERbeta expression in ovarian epithelial cancer primary tumors, and only ERalpha in metastatic tumors. Our findings indicate that a fundamental difference might exist between primary and metastatic cells, which could be caused by intrinsic or extrinsic factors that regulate ER gene expression.     

The effect of interferon gamma on epidermal growth factor receptor expression in normal and malignant ovarian epithelial cells.

OBJECTIVE: We examined the effect of interferon gamma on proliferation and epidermal growth factor receptor expression in ovarian cancer cell lines and normal ovarian epithelial cells. STUDY DESIGN: The tritiated thymidine incorporation assay was used to assess the effect of interferon gamma on proliferation. Scatchard analysis of anti-epidermal growth factor receptor antibody binding, and Western blotting of immunoprecipitates was used to assess the effect of interferon gamma on epidermal growth factor receptor expression. RESULTS: Although interferon gamma elicited 30% to 40% decreases in proliferation, epidermal growth factor receptor expression was strikingly increased in all four ovarian cancer cell lines. Scatchard analysis indicated that this increase occurred primarily at the cell surface, but total cellular receptor levels also were increased. In contrast, interferon gamma treatment of normal ovarian epithelial cells affected neither proliferation nor epidermal growth factor receptor levels. CONCLUSION: Because the up-regulation of epidermal growth factor receptors by interferon gamma appears to be confined to malignant cells, interferon gamma may facilitate immunotherapy and imaging of ovarian cancers by means of immunoconjugates directed against the epidermal growth factor receptor.     

Expression patterns in isoforms of vascular endothelial growth-factors in tissue samples of vulval cancer T1 N2M0 stage

OBJECTIVES: The aim of the study was estimating the expression of isoforms of mRNA vascular endothelial growth factor: VEGF121, VEGF145, VEGF165, VEGF183, VEGF189, VEGF206, in tissue samples of vulvar cancer, normal and inguinal lymph nodes metastases. MATERIALS AND METHODS: The material for this investigation of expression of mRNA selected genes was 4 tissue specimens from women with vulvar cancer T1N2M0. The method used for quantitative determination of the numbers of mRNA copies of selected genes was the QRT-PCR and QPCR technique, with the use of an ABI PRISM 7700 (TaqMan) sequence detector. The nucleotide sequence for starters and probes for quantitative RT-PCR was designed with the use of the Primer Express Version 1.0 ABI PRISM software. RESULTS: Expression of all mRNA isoforms VEGF was found in the samples. The number of mRNA copies obtained in 1 microgram total mRNA extract of the genes examined in tissue sections with various pathomorphology were clearly differentiated. CONCLUSIONS: Expression profile of aforementioned genes indicated a definitive relationship between vascular endothelial growth factor isoform and vulvar cancer. These findings provide evidence that the expression pattern of mRNA VEGF isoform should find broad applications in clinical and research settings.     

卵巢上皮性癌患者术前血清血管内皮生长因子与CA125水平的相关性及其预测预后的价值

目的分析卵巢上皮性癌（卵巢癌）患者术前血清血管内皮生长因子（VEGF）与CA125水平的相关性,探讨术前血清VEGF水平在卵巢癌患者预后判断中的价值。方法采用酶联免疫吸附试验（EHSA）测定41例卵巢癌患者（研究组）库存的术前血清中VEGF的水平,采用化学发光法测定同一份血清的CA125水平;以同期20例盆腔检查正常的妇女作为对照组。结合随诊资料,分析卵巢癌患者术前血清VEGF水平与CA125水平的相关性,并分析术前血清VEGF水平与患者复发和生存时间的关系。结果（1）研究组术前血清VEGF和CA125水平均明显高于对照组（VEGF中位数分别为415和165ng／L,CA125分别为611和16kU／L）,差异均有统计学意义（P〈0．01）;（2）Spearman等级相关分析显示,卵巢癌患者术前血清VEGF水平与血清CA125水平间无明显相关性（P=0．989）;（3）卵巢癌患者术前血清VEGF水平与其复发相关,复发者术前VEGF水平明显高于无复发者（中位数分别为490和315ng／L,P=0．035）;（4）单因素Kaplan-Meier法分析显示,卵巢癌患者术前血清VEGF水平与其生存时间呈负相关,高血清VEGF水平者的生存时间明显短于低血清VEGF水平者（中位数生存时间分别为18个月和〉35个月,P=0．010）;（5）多因素Cox回归模型分析显示,卵巢癌患者术前血清VEGF水平是与其生存时间有关的独立预后因素（P=0．042）。结论卵巢癌患者术前血清VEGF水平与CA125水平无明显相关性,VEGF水平变化是影响患者预后的独立因素。     

卵巢癌血管内皮生长因子过度表达及肿瘤浸润转移机制的探讨

目的　研究血管内皮生长因子 (VEGF)过度表达在卵巢癌浸润转移中的作用及可能机制。方法　脂质体介导VEGFcDNA转染卵巢癌细胞系CAOV3、COC1 ,并经逆转录聚合酶链反应 (RT PCR)技术、蛋白印迹 (Westernblot)法检测转染VEGFcDNA前后卵巢癌细胞中VEGF、明胶酶A(MMP 2 )mRNA及其蛋白表达水平 ;Boyden小室体外侵袭实验比较VEGFcDNA转染前后卵巢癌细胞穿透人工重组基底膜能力的变化。结果　VEGFcDNA转染后卵巢癌细胞系CAOV3、COC1细胞中VEGFmRNA表达水平分别为 0 98± 0 0 5和 0 87± 0 0 6 ,MMP 2mRNA表达水平分别为 0 95± 0 0 3和 0 82±0 0 2 ,均较转染前 (分别为 0 84± 0 0 3和 0 62± 0 0 4 ,0 71± 0 0 2和 0 61± 0 0 1 )明显增加 (P <0 0 5) ;VEGF、MMP 2蛋白表达水平也呈相同变化趋势 (P <0 0 5) ;VEGFcDNA转染后的CAOV3、COC1细胞的穿透人工重组基底膜的能力分别为 (42 5± 4 1 ) % ,(2 6 8± 2 4) % ,较对照组 [(2 4 7± 1 9) % ,(8 6±1 1 ) % ]明显增加 (P <0 0 5)。结论　VEGF促进卵巢癌浸润、转移的机制可能与诱导MMP 2合成及分泌有关。     

EGFR及MMP-9在上皮性卵巢癌组织中的表达及其与预后的关系

目的:研究表皮生长因子受体(EGFR)及基质金属蛋白酶-9(MMP-9)在上皮性卵巢癌中的表达,分析其与卵巢癌临床病理特征及预后的关系。方法:采用RT-PCR方法检测EGFR及MMP-9 mRNA在上皮性卵巢癌、卵巢良性肿瘤和正常卵巢组织的表达。结果:卵巢癌组织中EGFR和MMP-9 mRNA的阳性表达率分别为64.44%(29/45)、73.33%(33/45),相对表达水平为0.81±0.23、0.89±0.25,均明显高于卵巢良性肿瘤和正常卵巢组织,差异均有显著统计学意义(P均<0.05)。两者在有淋巴结转移的癌组织中的相对表达量均明显高于无淋巴结转移组,Ⅲ～Ⅳ期癌组织中的表达高于Ⅰ～Ⅱ期,差异均有统计学意义(P<0.01)。卵巢癌组织EGFR和MMP-9的表达呈显著正相关。Long-rank分析显示,EGFR和MMP-9共同表达患者的累积生存率明显低于阴性者,差异有显著统计学意义(P<0.05)。结论:上皮性卵巢癌组织中EGFR和MMP-9均呈现高表达,并且与卵巢癌的临床分期、淋巴结转移及患者的生存期密切相关,可作为判断上皮性卵巢癌恶性生物学行为和预后的重要参考指标。     

High expression of insulin-like growth factor binding protein–2 messenger RNA in epithelial ovarian cancers produces elevated preoperative serum levels

The molecular etiology of epithelial ovarian cancer remains unclear. Using microarray expression analysis, we recently reported that expression of the insulin-like growth factor binding protein-2 (IGFBP-2) gene is elevated in advanced epithelial ovarian cancers. The aim of this study was to further delineate the role of IGFBP-2 in the pathoetiology of epithelial ovarian cancer and determine if elevated ovarian cancer IGFBP-2 gene expression is reflected in serum. Relative IGFBP-2 expression was measured using quantitative real-time polymerase chain reaction in 113 epithelial ovarian cancers and 6 normal ovarian surface epithelial samples. Preoperative serum IGFBP-2 levels were measured by radioimmunoassay in 84 women (42 ovarian cancers, 26 benign gynecological conditions, and 10 healthy female controls). Ovarian cancers demonstrated 38-fold higher mean IGFBP-2 expression than normal ovarian epithelium (P < 0.01). Serum IGFBP-2 levels were elevated in women with early- and advanced-stage ovarian cancer compared to controls and patients with benign gynecological conditions (P = 0.05 and P < 0.01, respectively). Epithelial ovarian cancers express high levels of IGFBP-2 relative to normal ovarian epithelium, and this is associated with elevated serum IGFBP-2 levels compared to both normal controls and patients with benign gynecological disease. Our findings provide further support that the insulin-like growth factor pathway plays a significant role in epithelial ovarian cancer pathogenesis. Further, IGFBP-2 may represent an additional serum biomarker with utility in detection and monitoring of epithelial ovarian cancer.     

Relationship of hypoxia-inducible factor-1alpha expression and vascular endothelial growth factor in SKOV-3 ovarian cancer model

OBJECTIVE: To investigate the correlation of hypoxia-inducible factor (HIF)-1alpha and vascular endothelial growth factor (VEGF) or micro-vessel density (MVD). METHODS: The ovarian cancer cell line SKOV3 was transplanted into nude mice to form xenogeneic tumor. Mice were treated with rapamycin 4 mg/kg, sulindac 100 mg/(kg.d) and saline 200 microl respectively. Expression of HIF-1alpha and VEGF proteins and MVD were determined by immunohistochemistry. The mRNA of Glut1 and VEGF was studied by RT-PCR. RESULTS: The positive expression of HIF-1alpha and VEGF was moderate to strong, and MVD was high (31 +/- 8) in control group. In rapamycin treated group, the expression of HIF-1alpha was inhibited to weak positive or negative (P < 0.05), the VEGF protein and mRNA expression decreased (P < 0.05), MVD was also suppressed to a low level (13 +/- 5). There was a significant correlation between HIF-1alpha expression and VEGF expression (by Spearman's coefficient r = 0.8264, P < 0.01), and MVD (r = 0.4842, P < 0.05). CONCLUSIONS: HIF-1alpha protein can regulate VEGF protein and its mRNA expressions. It correlates with MVD in ovarian tumor tissue.