Isolation and purification of vascular endothelial growth factor (VEGF) from ascitic fluid of ovarian cancer patients

Vascular Endothelial Growth Factor (VEGF) or Vascular Permeability Factor (VPF) is an angiogenic cytokine expressed by many human and animal tumors. Because of the importance of VEGF in animal tumors, we purified VEGF/VPF from ascitic fluid of ovarian cancer patients with heparin sepharose column. The purified protein gave protein bands of 37 and 26 kD, respectively in 12% SDS PAGE. The specificity of the purified protein was determined with dot blot, trans-immunoblot and ELISA using polyclonal goat anti-VEGF antibody (Santa Cruz Biotechnology). The vasodilatatory effect of the purified protein was confirmed by a vascular permeability assay on mouse. A polyclonal mouse antibody was raised against the purified protein, which recognized the same protein by ELISA, transimmunoblot and dot-blot analysis. It has been also found that the raised polyclonal antibody in mouse- and the commercial VEGF polyclonal antibody (Santa Cruz Biotechnology) both inhibited in vitrocell proliferation of human MCF-7 cell line. This study shows for the first time an effort to purify VEGF from human source.(Pathology Oncology Research Vol 10, No 2, 104–108)     

卵巢癌的免疫治疗

卵巢癌作为妇女死亡的重要因素,一直受到人们的关注.手术和化学治疗目前都不能达到令人满意的效果.随着免疫学的进展,卵巢癌的免疫治疗将成为临床治疗的可选择手段之一.卵巢癌免疫治疗可分为预防性和治疗性免疫两种,同时还可分为腹腔内与全身两种给药方式.本文对卵巢癌的免疫治疗目前的一些进展,临床试验结果及发展中的一些问题与探索进行了综述.     

Influence of Human Ascitic Fluid on the In Vitro Antibacterial Activity of Moxifloxacin

Abstract

We investigated the in vitro influence of HAF on the antibacterial activity of moxifloxacin against Escherichia coli ATCC 10798, Escherichia coli K-12, Proteus rettgeri (Sanelli), Staphylococcus aureus ATCC 25923 Staphylococcus aureus NCTC 1808 and Staphylococcus epidermidis ATCC 12228. Human ascitic fluid was obtained from 6 cirrhotic patients by paracentesis. The interaction effect was evaluated by the checkerboard technique. Our results indicate the ability of human ascitic fluid to reduce minimum inhibitory concentrations of moxifloxacin against Gram-negative bacteria, but not against Gram-positives.     

乳酸脱氢酶水平在卵巢癌诊治中的意义

目的：探讨血清及腹水乳酸脱氢酶（ＬＤＨ）水平在卵巢癌诊治中的意义。方法：检测５０例卵巢癌及５５例其他妇科肿瘤术前血清ＬＤＨ水平,检测２０例腹水或盆腔冲洗液ＬＤＨ水平,并随访ＬＤＨ的变化,数据经计算机处理。结果：卵巢癌患者术前血清ＬＤＨ明显高于子宫恶性肿瘤和卵巢良性肿瘤（Ｐ〈０．００１）,晚期病人高于早期患者（Ｐ〈０．０５）,各病理类型之间无明显差异。卵巢癌患者腹水ＬＤＨ值远高于血清值,与良性肿瘤盆     

Screening for ovarian cancer

Ovarian cancer frequently presents late, when chances for long-term survival are poor. The increased survival advantage for patients diagnosed with early-stage ovarian cancer suggests that screening to detect early-stage disease might have an impact on disease mortality. Attempts are being made to develop effective screening procedures for early ovarian cancer in symptom-free women, using a variety of serum tumor markers, proteomic patterns, and ovarian morphological and vascular features. Two distinct screening strategies have emerged, one utilizing transvaginal scanning as the primary test, and the other involving measurement of the serum tumor marker CA125 as the primary test with transvaginal ultrasonography as the secondary test (multimodal screening). Large randomized trials are now underway to provide definitive data on the impact of screening on mortality and address morbidity, health economics and psychosocial issues.     

The possible role of factor H in colon cancer resistance to complement attack

A soluble complement inhibitor factor H (FH) and its splice variant factor H-like protein (FHL) have been recently discovered to play a major role in malignant cell escape from complement-mediated cytotoxicity in lung-, ovarian- and glia-derived neoplasms. The role of FH in colon cancer has not yet been examined. Here, we studied immunocytochemically FH/FHL expression in tumor samples derived from 40 patients, with both primary colon adenocarcinoma and metastatic foci in the liver. FH/FHL immunoreactivity was present in stroma of both primary and metastatic tumors, in virtually all patients. The cellular immunoreactivity was observed infrequently. Importantly, when analyzed quantitatively, FH/FHL immunoreactivity was significantly increased in liver metastases when compared with the primary sites. In addition, we have analyzed FH and FHL expression in 5 colon cancer cell lines: SW480, SW620, HCT116, HT-29 and Lovo. FH mRNA and FH secretion were observed in SW620 and HT-29 cells, whereas FHL was produced only by HT-29 cell-line. By confocal and electron microscopy, FH immunoreactivity was associated with the plasma membrane and intracellular vesicular structures. Finally, we have analyzed the role of FH in the susceptibility of SW620 colon cancer cells to complement-mediated damage. When FH function was blocked, using specific antibody, the cells became more susceptible to lysis. Taken together, our results suggest an important role of FH/FHL in colon cancer cells defense against complement-mediated cytotoxicity, and in metastatic process.     

吉西他滨加顺铂治疗复发性卵巢癌

目的:评估吉西他滨联合顺铂治疗复发性卵巢癌的疗效及毒性。方法:28例复发性卵巢癌,用吉西他滨1000mg/m2和顺铂35mg/m2,静注,第1、8天,21天为1周期。结果:28例患者总有效率60.7%(95%可信区间41.7%-79.6%),其中CR5例(17.9%),PR12例(42.9%)。中位疾病进展时间5.5个月(2.5~20个月),中位生存期12.5个月。其中16例铂类耐药和12例铂类敏感患者的有效率、中位生存期分别为56.3%和66.7%(P=0.95)、10.5和14.5个月(P=0.003)。毒性主要是白细胞减少和血小板减少。结论:吉西他滨加顺铂是治疗复发性卵巢癌的有效方案,不仅可用于铂类敏感患者,也可用于铂类耐药患者。其毒性可以接受。     

Tumor infiltrating lymphocytes in ovarian cancer

The accumulation of tumor infiltrating lymphocytes (TILs) in ovarian cancer is prognostic for increased survival while increases in immunosuppressive regulatory T-cells (Tregs) are associated with poor outcomes. Approaches that bolster tumor-reactive TILs may limit tumor progression. However, identifying tumor-reactive TILs in ovarian cancer has been challenging, though adoptive TIL therapy in patients has been encouraging. Other forms of TIL immunomodulation remain under investigation including Treg depletion, antibody-based checkpoint modification, activation and amplification using dendritic cells, antigen presenting cells or IL-2 cytokine culture, adjuvant cytokine injections, and gene-engineered T-cells. Many approaches to TIL manipulation inhibit ovarian cancer progression in preclinical or clinical studies as monotherapy. Here, we review the impact of TILs in ovarian cancer and attempts to mobilize TILs to halt tumor progression. We conclude that effective TIL therapy for ovarian cancer is at the brink of translation and optimal TIL activity may require combined methodologies to deliver clinically-relevant treatment.     

Novel target antigens for dendritic cell-based immunotherapy against ovarian cancer

Identification of tumor-specific target antigens has been a major hurdlefor the treatment of malignant disease by vaccination or immunotherapy. A second challenge has been the induction of therapeutically effective immune responses to these ‘self’ antigens. The recent recognition of dendritic cells as powerful antigen-presenting cells capable of inducing primary T-cell responses in vitro and in vivo – in combination with identification of tumor-specific antigens – has generated widespread interest in dendritic cell-based immunotherapy against a wide variety of tumors. In this review, a series of recently identified novel ovarian tumor antigens is discussed, and the potential for therapeutic dendritic cell vaccination targeted against these antigens is assessed.     

顺铂和依托泊苷及六甲蜜胺联合治疗铂类敏感复发性卵巢癌的临床分析

目的:探讨顺铂(DDP)、依托泊苷(Vp16)和六甲蜜胺(HMM)联合化疗治疗铂类敏感复发性卵巢癌的临床疗效。方法:对48例距前次铂类化疗6个月以上复发的卵巢癌患者用DDP+Vp16+HMM化疗,其中21例直接用该方案化疗,27例行再次减瘤术后再用此方案化疗。具体方案:DDP60mg/m2,静脉滴入,d1;Vp1650mg/d,口服,d1～d14;HMM200mg/d,口服,d15～d28。28d为1个疗程。结果:化疗中位疗程数5个。非手术组中19例可评价化疗效果,完全有效6例,部分有效8例,总有效率73.7%(95%CI48.8%～90.9%)。全组42例患者可评价生存期,中位肿瘤非进展生存期(progressionfreesurvival,PFS)和总生存期(overallsurvival,OS)分别为9.5和19.6个月。其中理想手术(15例)、亚理想手术(8例)、非手术(19例)组患者的中位PFS和OS分别为13.5和28.4个月、8.5和14.0个月、8.8和17.3个月。理想减瘤术患者的PFS和OS均明显优于亚理想手术(P值分别为0.0117和0.0083)和非手术的患者(P分别为0.0027和0.0054)。该方案主要毒副反应是恶心呕吐,Ⅲ～Ⅳ度发生率达62.5%,其中10例患者因此而中断化疗。血液学毒性和肾毒性较轻。结论:DDP+Vp16+HMM是治疗铂类敏感复发性卵巢癌的有效方案之一,其主要毒副反应是恶心呕吐,通过控制多数患者可以耐受。理想的再次减瘤术可以改善患者的生存期。     

年轻妇女卵巢上皮癌的临床特点及预后分析

背景与目的:卵巢上皮癌多发生于老年妇女,年轻妇女较少见.有关35岁以下妇女卵巢上皮癌的临床特点、预后因素分析报道较少.本研究旨在探讨年轻妇女卵巢上皮癌的临床特点、治疗、生存率及预后因素分析.方法:回顾性分析1980年1月至2003年12月我院收治的71例≤35岁的卵巢上皮癌患者的临床资料.生存率用寿命表法计算.利用Cox模型分析比较影响预后的因素.结果:71例确诊为卵巢上皮癌患者中位年龄28岁.临床表现为自扪及腹部包块或体检发现腹部包块18例,腹痛、腹胀各11例.肿物最大径平均为13.7 cm,肿瘤位于单侧52例(73.2%),68例(95.7%)行满意细胞减灭术,手术病理分期I期44例(62.O%)、Ⅱ期5例、Ⅲ期18例、Ⅳ期4例.病理类型以浆液性囊腺癌(40例,56.3%)和粘液性囊腺癌(22例,30.9%)为最多.病理分级为高分化42例(59.2%)、中分化18例(25.4%)、低分化11例(15.5%).68例术前或术后进行了以铂类和紫杉醇类为基础的化疗.15例保守手术中(均为I a、G1期患者),12例无瘤生存(80.0%).按寿命表法计算的2年生存率为86.0%.5年生存率为82.0%.Cox模型多因素分析显示病理分级、残留病灶大小是影响年轻妇女卵巢上皮癌预后的因素(P<0.05).结论:35岁以下妇女卵巢上皮癌患者,以单侧多见,以浆液性囊腺癌多见,预后好.部分I a、G1期患者可保留生育功能.病理分级、残留病灶大小是影响35岁以下妇女卵巢上皮癌预后的因素.     

肿瘤细胞裂解物致敏的树突状细胞对大鼠卵巢上皮癌免疫治疗的体内研究

目的：研究树突状细胞（dendritic cells,DC）与肿瘤细胞裂解物致敏的树突状细胞（tumor-associated antigen／dendritic cells,TAA／DC）对荷瘤大鼠的免疫治疗效果。方法：建立大鼠卵巢癌皮下移植瘤模型;于成瘤后2周分别用PBS、DC或TAA／DC进行免疫治疗,比较治疗效果。结果：TAA／DC抑制肿瘤细胞的增殖与转移,对荷瘤大鼠治疗后8周,局部肿瘤无明显增长,P=0．355,无转移灶出现;TAA／DC可明显改善大鼠免疫状况,对荷瘤大鼠治疗后3周,使已经下降的CD8^＋ T细胞百分比又明显上升,P=0．000;TAA／DC对荷瘤大鼠单次免疫治疗的效果具有时限性,在对荷瘤大鼠治疗后8周,CD8^＋ T细胞百分比再次出现明显下降,P=0．000。结论：TAA／DC可改善荷瘤大鼠的细胞免疫状态,抑制肿瘤细胞的增殖与转移。     

Understanding dendritic cell immunotherapy in ovarian cancer

Introduction: Approximately eighty percent of patients with ovarian cancer are diagnosed with advanced disease. Even with cutting edge surgical techniques and the best regimens of standard therapies most patients relapse and die of drug resistant disease within five years of diagnosis. Dendritic cell (DC) immunotherapy can induce anti-tumor T cell immunity in patients and holds great potential in the era of modern anti-cancer treatment.

Areas Covered: This review outlines critical factors regulating the outcome of DC immunotherapy in ovarian cancer, summarizes the important findings in ovarian cancer DC clinical trials, and discusses new directions which may improve the effectiveness of DC immunotherapy.

Expert Commentary: Administration of DC vaccines with other forms of immunotherapy may enhance the efficacy of these treatments, ultimately increasing cures for this disease.     

卵巢上皮癌治疗后远处转移的影响因素

目的：探讨卵巢上皮癌治疗后远处转移的影响因素。方法：对我院1990年1月5日～1995年12月10日收治的40例卵巢上皮癌远处转移者进行回顾分析。结果：初次手术后残余癌灶≤2cm和〉2cm者远处转移率分别为32．5％和60％。全身化疗〈4个疗程和≥4个疗程者转移率分别为62．5％和37．5％，有腹水者和无腹水者的转移率分别为65％和36％P〈0．05。结论：卵巢上皮癌初次治疗后残余癌灶大小、化疗疗程、有恶性腹水者是远处转移的重要因素。     

卵巢上皮癌治疗后远处转移的影响因素

目的:探讨卵巢上皮癌治疗后远处转移的影响因素。方法:对我院1990年1月5日～1995年12月10日收治的40例卵巢上皮癌远处转移者进行回顾分析。结果:初次手术后残余癌灶≤2cm和>2cm者远处转移率分别为32.5%和60%。全身化疗<4个疗程和≥4个疗程者转移率分别为62.5%和37.5%,有腹水者和无腹水者的转移率分别为65%和36%P<0.05。结论:卵巢上皮癌初次治疗后残余癌灶大小、化疗疗程、有恶性腹水者是远处转移的重要因素。     

High tumor tissue concentration of urokinase plasminogen activator receptor is associated with good prognosis in patients with ovarian cancer

The urokinase plasminogen activator (uPA) system is involved in tumor growth and metastasis. We assayed the components of the uPA system in homogenates of 64 primary epithelial ovarian tumors and 5 metastases and evaluated the association of these parameters to prognosis in the 51 malignant cases. The levels of uPA, PAI-2 and the uPA:PAI-1 complex increased with progressive loss of histological differentiation (p(trend) <0.001, <0.05 and <0.001). The level of PAI-1 was higher in poorly than in well/moderately differentiated tumors (p = 0.03). The content of uPAR was lower in benign tumors as compared to borderline malignancies (p = 0.002), invasive primary tumors (p < 0.001), and metastases (p = 0.002). Surprisingly, the level of uPAR was lower in poorly differentiated as compared to both borderline (p = 0.01) and well differentiated malignant tumors (p = 0.005). Also, the level of uPAR was lower in advanced as compared to early stages of the disease (p(trend) = 0.002). The median follow-up time for patients was 5.8 years. High tumor tissue levels of uPAR were associated with longer postoperative survival (HR = 0.4, 95% CI = 0.2-0.8, p = 0.01). In contrast, shorter survival was evident in patients with high tumor levels of uPA from 2 years on after operation (HR = 4.6, 95% CI = 1.2-17, p = 0.02). High tPA levels tended to be associated with shorter overall survival after 2 years (HR = 2.9, 95% 95% CI = 0.9-9.8, p = 0.08). Although high tumor tissue content of uPAR was associated with a less aggressive phenotype characterized by well differentiated histology and longer survival, low content of uPAR in the poorly differentiated tumors and metastases presumably results from increased elimination of uPAR.     

异环磷酰胺联合自体PHA-LAK细胞治疗晚期复发卵巢上皮癌的疗效观察

目的研究异环磷酰胺(ifosfamide,IFO)联合自体PHA-LAK细胞过继性免疫治疗晚期复发卵巢上皮癌的疗效。方法25例复发的晚期卵巢上皮癌患者经IFO1200mg/(m2·d)连用4d,同时于用IFO后0、4、8h分别静脉推注美司钠400mg;化疗间歇期以PHA-LAK细胞治疗4次。联合治疗2个周期以上。结果25例患者中,完全缓解(CR)3例,部分缓解(PR)8例,稳定(NC)7例,进展(PD)7例,有效率(CR+PR)44%(11/25),中位无进展生存期(TTP)21周,中位生存期(OS)47周。毒副反应主要是消化道反应和骨髓抑制。结论IFO联合自体PHA-LAK细胞治疗复发的晚期卵巢上皮癌有效,毒副反应低,可作为晚期复发卵巢上皮癌的选择方案。     

CSD多肽抑制卵巢癌SKOV3细胞的侵袭和迁移

目的:研究caveolin-1脚手架结构域（caveolin-1 scaffolding domain,CSD）多肽对卵巢癌SKOV3细胞迁移和侵袭能力的影响,并探讨其分子机制。方法:用细胞免疫荧光法检测CSD多肽在SKOV3细胞中的渗透情况,划痕实验检测细胞的迁移能力,Transwell细胞侵袭实验检测细胞的侵袭能力,用细胞免疫荧光和蛋白免疫印迹法检测E-cadherin和Vimentin蛋白的表达,蛋白免疫印迹法检测SKOV3细胞中EGFR、PI3K、Akt蛋白的相对磷酸化水平。结果:细胞免疫荧光显示CSD多肽能够顺利进入SKOV3细胞内,与空白对照组相比,CSD多肽能明显抑制SKOV3细胞的迁移能力（P＜0.01）和侵袭能力（P＜0.01）,并且CSD多肽能增加细胞中E-cadherin蛋白水平（P＜0.01）和下调Vimentin蛋白的表达（P＜0.01）;SKOV3细胞用CSD多肽处理后,EGFR、PI3K、Akt蛋白的相对磷酸化水平明显低于空白对照组（P＜0.01,P＜0.05,P＜0.05）。结论:CSD多肽在体外能抑制SKOV3细胞的侵袭和迁移能力,逆转上皮-间质转化表型,EGFR/PI3K/Akt信号通路活性水平的下降可能是其重要的分子机制。     

TM4SF1 蛋白在上皮性卵巢癌组织中的表达及其临床意义

目的 探讨跨膜四超家族成员1（transmembrane 4 super family 1,TM4SF1）蛋白在人卵巢组织中的表达及其与上皮性卵巢癌临床病理因素的关系。方法 采用免疫组化SP法检测16例正常卵巢上皮组织、23例上皮性卵巢良性肿瘤组织及55例上皮性卵巢癌癌组织中TM4SF1蛋白的表达情况,分析其与上皮性卵巢癌临床病理因素及预后的关系。结果 ①TM4SF1蛋白在上皮性卵巢癌癌组织中的阳性表达率（90.90%）高于上皮性卵巢良性肿瘤组织（65.22%）及正常卵巢上皮组织（25.00%）的阳性表达率,差异有统计学意义（P〈0.05）。②TM4SF1蛋白在正常卵巢上皮细胞中的表达主要位于上皮细胞的细胞膜和细胞膜近基底膜处及细胞质中,在卵巢良性肿瘤组织中的表达主要集中在肿瘤细胞的细胞膜近基底膜处及细胞膜其他部位,而在卵巢癌细胞中则主要分布于癌细胞的细胞质中。③Ⅲ～Ⅳ期卵巢癌患者TM4SF1蛋白的阳性表达率高于Ⅰ～Ⅱ期患者,中低分化癌患者TM4SF1蛋白的阳性表达率高于高分化癌患者（P〈0.05）。④TM4SF1蛋白的阳性表达率与组织学类型、腹水量的多少无明显相关（P〉0.05）,而与FIGO分期及组织学分级明显相关（P〈0.05）。⑤Cox比例风险回归模型多因素分析显示TM4SF1蛋白在卵巢癌癌组织中的阳性表达不是影响患者生存的独立预后因素。结论 TM4SF1蛋白在上皮性卵巢癌癌组织中呈高表达,可能与卵巢癌的发生、发展相关。