肝素结合表皮生长因子在卵巢癌中的研究进展

肝素结合表皮生长因子(HB-EGF)是表皮生长因子家族成员之一,可通过激活表皮生长因子受体发挥生物学效应。近来研究表明,HB-EGF在卵巢癌中高水平表达并在肿瘤的发生发展中起重要作用,因此成为极具潜力的卵巢癌治疗靶点。系统描述HB-EGF的特性及其特异性抑制剂白喉毒素突变体(CRM197)的研究现状,并讨论CRM197分别和膜型基质金属蛋白酶(MTl-MMP)抑制剂、去整合素-金属蛋白酶(ADAM)抑制剂及紫杉醇联合治疗卵巢癌的发展潜力。     

卵巢癌顺铂耐药细胞系的建立及耐药机制的研究

顺铂 (ｃｉｓｐｌａｔｉｎ ,ＤＤＰ)是一种有效的抗肿瘤药物 ,耐药的产生极大地影响卵巢癌患者对化疗的敏感程度 ,限制了其临床疗效[1] 。本研究采用中等浓度、间歇作用[2 ] 的方法 ,于 1998年 12月到 1999年 5月建立了人卵巢癌细胞株ＳＫＯＶ3对顺铂耐药细胞系ＳＫＯＶ3/ＤＤＰ ,对耐药指数、细胞周期分布、细胞内谷胱甘肽 (ＧＳＨ )含量、拓扑异构酶Ⅱ(ＴＯＰⅡ )含量及活性进行检测 ,从细胞水平、分子生物学水平对其基本特征及耐药机制进行研究。1　材料与方法1 1　研究资料　人卵巢癌细胞株ＳＫＯＶ3购自北京医科大学妇科教研室 ,细胞株…     

交叉反应物质197在卵巢癌耐药中的研究进展

卵巢癌起病隐匿,多数晚期卵巢癌患者会出现复发及对常规化疗耐药现象,严重威胁女性的生命健康。因此,化疗耐药已成为治愈卵巢癌的关键制约因素。肝素结合表皮生长因子(HB-EGF)作为表皮生长因子家族成员,已经被证实是一种有前景的卵巢癌化疗靶点。交叉反应物质197(cross-reacting material 197,CRM197)是一种无毒的白喉毒素变异体,特异性结合并抑制HB-EGF,从而抑制卵巢癌化疗耐药。研究表明,CRM197在多种肿瘤治疗及卵巢癌耐药方面均有显著作用,但是关于CRM197参与卵巢癌耐药的具体机制仍有待进一步研究。本文系统性描述CRM197的特性,并讨论CRM197在肿瘤治疗中的应用及CRM197与顺铂、紫杉醇联合应用在卵巢癌耐药中的研究进展。     

卵巢癌顺铂耐药的分子机制研究进展

顺铂耐药是卵巢癌术后复发和预后不良的重要原因之一.卵巢癌的顺铂耐药机制复杂,除抑癌基因、癌基因、耐药相关基因的改变外,还涉及染色体改变、细胞解毒过程及DNA损伤修复能力的增强以及细胞骨架、热休克蛋白表达异常等,对其分子机制的了解有助于指导卵巢癌临床治疗方案的选择.     

核苷酸切除修复与卵巢癌顺铂耐药

DNA操作修复能力增强是导致卵巢顺铂耐药的主要原因之一。由铂-DNA加合物引起的DNA损伤修复主要通过核苷酸切除修复途径，该签字径涉及多种蛋白，包括损伤识别、切开、切除、修复合成和DNA连接等5个步骤，大量基础和临床研究表明核苷酸切除修复基因表达水平的增加与卵巢癌顺铂耐药相关联。应用基因治疗、化疗增效液晶或联合化疗抑制肿瘤细胞的DNA修复能力将增加卵巢癌患者顺铂敏感性。     

核苷酸切除修复与卵巢癌顺铂耐药

DNA损伤修复能力增强是导致卵巢癌顺铂耐药的主要原因之一,由铂-DNA加合物引起的DNA损伤修复主要通过核苷酸切除修复途径,该途径涉及多种蛋白,包括损伤识别、切开、切除、修复合成和DNA连接等5个步骤.大量基础和临床研究表明核苷酸切除修复基因表达水平的增加与卵巢癌顺铂耐药相关联.应用基因治疗、化疗增效剂或联合化疗抑制肿瘤细胞的DNA修复能力将增加卵巢癌患者顺铂敏感性.     

胰岛素样生长因子1受体与卵巢癌SKOV3细胞顺铂耐药相关性的研究

目的:探讨胰岛素样生长因子1受体(IGF-1R)与卵巢癌SKOV3细胞顺铂耐药的相关性。方法:建立人卵巢癌细胞系SKOV3异种移植瘤模型,其中部分用顺铂处理;由此模型获得细胞分为SKOV3-P组(未化疗组)和SKOV3-R组(化疗组)。采用免疫细胞化学法和流式细胞术检测瘤细胞中IGF-1和IGF-1R的表达差异;MTT法和流式细胞术分别检测不同浓度顺铂(0、1、5、10、15、20μg/ml)和IGF-1R抑制剂NVP-AEW541(0、1、5、10、20μmol/L)单独及联合应用对瘤细胞的增殖及凋亡的影响。结果:SKOV3-P组、SK-OV3-R组细胞中均表达IGF-1和IGF-1R,且SKOV3-R组细胞的表达均显著高于SKOV3-P组细胞(P<0.05);NVP-AEW541对SKOV3-P组、SKOV3-R组细胞均表现增殖抑制,呈现剂量-时间依赖关系;NVP-AEW541联合顺铂作用瘤细胞时抑制作用显著增强,移植瘤细胞的顺铂敏感性显著增加(P<0.05);流式细胞术检测联合用药组细胞(10μmol/LNVP-AEW541+10μg/ml顺铂)凋亡率明显高于对照组和单独用药组(10μmol/L NVP-AEW541),且SKOV3-P组细胞的抑制率和凋亡率均显著高于SKOV3-R组细胞(P<0.05)。结论:IGF-1R参与了化疗耐药过程,通过NVP-AEW541抑制IGF-1R可能逆转卵巢癌细胞的顺铂耐药。     

肝素结合性表皮生长因子与生殖

肝素结合性表皮生长因子（ＨＢ－ＥＧＦ）是近年来发现的属表皮生长因子（ＥＧＦ）超家族中的新成员,其特点是对肝素有较强的亲和性,可与细胞表面的ＥＧＦ受体（ＥＧＦＲ）和硫酸乙酰肝素蛋白多糖（ＨＳＰＧ）两种受体相结合而发挥其重要的生物学作用;其本身也是一种旁分泌,自分泌生长因子和粘附分子。     

脱氧氟尿苷逆转人卵巢癌细胞系顺铂耐药的研究

脱氧氟尿苷(FUDR)是高效低毒的新一代氟尿嘧啶类抗肿瘤药物,尚未见有关FUDR用于逆转肿瘤耐药的报道。本研究采用四甲基偶氮唑蓝(MTT)法、原子吸收分光光度法、免疫组化SP法等方法,分析了FUDR对卵巢癌细胞系SKOV3/DDP顺铂敏感性的影响,并初步探讨其逆转机制,为临床耐药逆转提供理论依据。     

PARP抑制剂在铂耐药上皮性卵巢癌治疗中的应用

PARP抑制剂在上皮性卵巢癌的一线维持治疗中展示出了良好的临床效果,但在铂耐药上皮性卵巢癌的治疗中尚无较多临床数据提供参考。本文对PARP抑制剂在铂耐药上皮性卵巢癌治疗中的应用做一综述,以期提供临床参考。     

Wnt通路抑制因子甲基化与卵巢癌耐药的相关性

目的探讨Wnt通路抑制因子甲基化与卵巢癌耐药的相关性。方法2008年在四川大学华西第二医院采用甲基化特异性PCR(MSP)、RT-PCR检测卵巢癌细胞株SKOV3、OVCAR-8(紫杉醇耐药),A2780及A2780/D(顺铂耐药)中,Wnt通路抑制因子SFRP-1、2、4、5,WIF-1、DKK-3的甲基化状态、mRNA水平;四甲基偶氮唑蓝(MTT)法比较DNA甲基转移酶抑制剂5-氮-2-脱氧胞苷(5-Aza-CdR)处理后,OVCAR-8细胞耐药逆转的情况,并检测WIF-1基因甲基化状态、mRNA表达水平的改变。结果WIF-1基因在OVCAR-8细胞中为高甲基化表达抑制,5-Aza-CdR可使其去甲基化后重新表达,并部分逆转OVCAR-8细胞的紫杉醇耐药。结论WIF-1基因的高甲基化状态与卵巢癌耐药细胞株OVCAR-8的紫杉醇耐药相关,去甲基化后可部分逆转耐药,这为逆转卵巢癌耐药提供了新的研究方向。     

Expression of the opioid growth factor-opioid growth factor receptor axis in human ovarian cancer

Objective

The opioid growth factor (OGF) and its receptor (OGFr), serve as inhibitory axis regulating cell proliferation in normal cells and cancer. We investigated the presence and relative expression of OGF and OGFr in normal human ovarian surface epithelial (HOSE) cells, benign ovarian cysts, and ovarian cancers.

Methods

Surgical samples of 16 patients with ovarian cancer and 27 patients with ovarian benign cysts were obtained intraoperatively. HOSE were collected by scraping the surface of normal ovaries of 10 post menopausal women undergoing hysterectomy and oophorectomy. Semiquantitative immunohistochemistry was used to assess the presence, distribution, and levels of OGF and OGFr. Receptor binding assays measured binding capacity and affinity of OGFr for radiolabeled OGF.

Results

OGF and OGFr were present in HOSE cells, ovarian cysts, and ovarian cancers. Compared to HOSE cells, OGF and OGFr protein levels were reduced 29% and 34% (p < 0.001), respectively, in ovarian cysts, and decreased 58% and 48% (p < 0.001), respectively, in ovarian cancers. Binding assays revealed 5.4 fold fewer OGFr binding sites in cancers than cysts (p < 0.05). Levels of OGF and OGFr were comparable in primary, metastatic, or recurrent ovarian cancers.

Conclusion

We have shown that a native opioid pathway, the OGF-OGFr axis, is present in human ovarian cancer. Importantly, the expression of OGF and OGFr is diminished in human ovarian cancer. As OGF and OGFr normally function in maintaining cell proliferation, therapy to harness OGF/OGFr function could provide a useful biologic-based treatment for human ovarian cancer.     

Increased circulating hepatocyte growth factor (HGF): a marker of epithelial ovarian cancer and an indicator of poor prognosis

Objective

Hepatocyte growth factor (HGF) has been described to be increased in different cancers. In the present study we wanted to investigate whether HGF in serum can distinguish between benign and malignant ovarian tumors, and whether serum HGF levels can predict the outcome in patients with ovarian carcinomas.

Methods

We included 123 consecutive patients appointed for laparotomy due to a pelvic mass. Preoperative levels of serum cancer antigen 125 (CA 125), HGF and HGF activator (HGFA) were quantified with immunological methods. We performed immunohistochemical analyses of HGFα, HGFβ and the receptor c-Met. Five-year survival of patients with advanced disease (stage III and stage IV) was analyzed with the Kaplan-Meier method.

Results

Sixty patients had ovarian carcinomas, 23 borderline tumors, and 40 benign ovarian tumors. Patients with ovarian carcinomas had significantly higher preoperative HGF and CA 125 serum levels than patients with benign ovarian tumors, and borderline tumors. Patients with borderline tumors had significantly higher CA 125 values than benign cases. A combination of CA 125 and HGF increased the specificity in predicting carcinoma. We observed abundant HGFα, HGFβ and c-Met expressions in all ovarian tumors. Patients with advanced disease and preoperative serum HGF values ≥ 2 SD above reference value had a shorter disease-free survival than patients with advanced disease and serum HGF < 2 SD above reference value.

Conclusions

HGF in serum is an indicator of ovarian carcinoma in women with a pelvic mass, and of a poor prognosis in advanced ovarian cancer.     

Expression of glutathione S-transferase π and cisplatin resistance in human ovarian cancer

Glutathione S-transferase π (GSTπ) expression in tumor cells is thought to relate to cisplatin resistance. We attempted to clarify immunohistochemically the correlation between GSTπ expression and clinical response in ovarian cancer. Fifty-nine patients with ovarian cancer underwent initial debulking surgery and received from three to five courses of cisplatin-based chemotherapy after surgery. Immunostaining for GSTπ was performed on formalin fixed sections of the patients' tumor by the streptoavidin-biotin-peroxidase complex method. Positive staining of GSTπ was observed diffusely in nuclei and cytoplasm of cancer cells. Of 59 tumors, 38 (64.4%) were GSTπ positive. Twenty-three of 25 patients (92.0%) who showed no response to chemotherapy had GSTπ positive tumor cells. The predictive value of positive GSTπ staining for drug resistance was 79.3% (23/29). The 5-year survival rate of patients with GSTπ positive tumors was significantly lower than that of those with GSTπ negative tumors by the Kaplan-Meier method ( P < 0.01). The results suggest that overexpression of GSTπ is related to resistance to cisplatin and to prognosis in patients with ovarian cancer.     

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。