具生殖系嵌合能力的C57BL/6J小鼠胚胎干细胞系的建立

目的:建立能实现种系传递的C57BL/6J小鼠胚胎干细胞系。方法:从C57BL/6J小鼠3.5d的囊胚中分离培养内细胞团。经体外适宜培养建系后,将C57BL/6J胚胎干细胞(ES)注入ICR小鼠受体囊胚腔,制备嵌合体小鼠。结果:成功地建立了3个C57BL/6J小鼠胚胎干细胞系,该C57BL/6JES细胞呈集落状生长,正常稳定的核型率>80%,具高水平的磷酸酶活性,并表达ES细胞特殊性细胞表面标记SSEA-1,不表达SSEA-3和SSEA-4;体内外的分化实验也证实mC57ES1具向三胚层组织分化的能力。经显微注入ICR小鼠囊胚腔中,产生4只嵌合体小鼠,经证实其中1只为种系嵌合小鼠。结论:建立了能实现种系传递的C57BL/6J小鼠胚胎干细胞系,该系的ES细胞可用于今后制备转基因动物和基因敲除动物。     

Fischer 344大鼠上皮性卵巢癌动物模型的建立及其生物学特性

目的 :在具有正常免疫功能的Fischer3 44大鼠体内建立卵巢上皮癌模型并探讨其生物学特性。方法 :体外培养近交系Fischer3 44大鼠卵巢上皮低分化腺癌细胞株NuTu 1 9,采用细胞计数和MTT法观察细胞生长情况及其对顺铂 (cDDP) ,5 氟尿嘧啶 (5 FU)和足叶乙甙 (VP 1 6)的敏感性。将对数生长期的NuTu 1 9按 1× 1 0 7、1× 1 0 6和 1× 1 0 5细胞 /只接种于雌性Fischer3 44大鼠腹腔内 ,建立卵巢癌腹腔转移模型 ,观察成瘤率、肿瘤生长、播散情况及动物生存期 ,并对腹腔肿瘤进行病理学检查和原代组织培养 ,确定肿瘤的存在及性质。结果 :在体外培养条件下NuTu 1 9细胞生长迅速 ,其倍增时间约 1 8.9h。NuTu 1 9对cDDP和VP 1 6等卵巢癌常用化疗药物高度敏感。将不同数量的NuTu 1 9细胞接种大鼠腹腔后 ,成瘤率为 1 0 0 % ,迅速发生腹腔播散并产生大量血性腹水 ,其生物学行为与人类卵巢上皮癌相似。肿瘤细胞负荷增加 ,动物生存期明显缩短 (P <0 .0 1 )。模型动物肿瘤组织病理学检查证实胸腹腔多脏器肿瘤播散 ,腹腔肿瘤原代培养证实肿瘤组织的细胞形态和生物学特性与NuTu 1 9细胞一致。结论 :在具有正常免疫功能的大鼠体内建立卵巢上皮癌动物模型 ,有助于了解卵巢癌生物学及免疫学特性 ,可作为良好的肿瘤模型用于卵巢     

三种人卵巢癌动物模型的生物学特性比较

目的:建立人卵巢癌裸小鼠皮下移植瘤,腹水瘤和原位移植瘤模型,观察比较其生物学特性,为卵巢癌的理论研究和选择临床模型选择提供参考。方法:利用卵巢癌细胞株SW626先制备裸小鼠皮下移植瘤模型,再将瘤组织条植入裸小鼠一侧卵巢内,建立人卵巢癌的原位移植模型,将该细胞悬液注入小鼠腹腔建立腹水瘤模型。用组织病理学,电镜和流式细胞仪DNA含量及染色体核型分析鉴定模型,并比较3种模型在生物学特性上的差异。结果:3种模型的瘤细胞与细胞株在形态和结构上一致,腹水瘤模型自然生存率明显短于其它两种模型,后两者无明显差异。在生物学特性上各具特点,其中原位移植瘤模型中肿瘤的生长和转移完全模拟人卵巢癌的临床过程。结论:3种模型从不同层面展现了人卵巢癌的生物学特点,其中原位移植瘤模型是研究卵巢癌的转移基础和评定抗癌药物药效更客现的模型。     

人卵巢癌裸鼠原位移植瘤模型的建立及其生物学特性研究

随着肿瘤实验动物学的发展，人们对肿瘤模型的仿真性日益重视。人肿瘤原位移植瘤模型可以客观地再现人类肿瘤的临床发展过程，自然模拟肿瘤在人体内的浸润和转移过程。我们利用卵巢癌细胞株SW626成功地建立了人卵巢癌裸鼠的原位移植瘤模型，并对其生物学特性进行了研究，现报道如下。     

人绒毛膜癌鼠肺转移模型的建立及其生物学特性的初步观察

目的 建立1种较为符合临床特征的人绒毛膜癌(绒癌)肺转移动物模型,初步观察其生物学特性,并探索成功建立模型的合适的细胞浓度、细胞数量.方法 将40只5～6周龄重症联合免疫缺陷(SCID)小鼠随机分为4组,每组各10只,人绒癌细胞系JEG-3分别以1×107个细胞/ml×0.1 ml(A组)、5×106个细胞/ml×0.2 ml(B组)、1×106个细胞/ml×0.1 ml(C组)注入SCID小鼠尾静脉,另设对照组.接种后每3天观察1次小鼠状态,称量1次小鼠的体质量,绘制小鼠体质量变化曲线;当小鼠处于濒死状态时,用小动物成像CT(Micro CT)系统检查肺部有无转移灶、转移灶数目及大小.Micro CT检查后,解剖SCID小鼠,取肺部转移灶进行细胞的原代培养并行病理取材,HE染色、病理检查,免疫组织化学方法检测人绒毛膜促性腺激素β亚单位(β-hCG)的表达情况;原代培养细胞行免疫组织化学检测β-hCG的表达情况,并行染色体核型分析.结果 A组小鼠接种时全部死亡;C组接种时死亡2只,存活的8只在(30.0±2.0)d时,处于濒死状态,进行解剖未发现肺转移灶.B组接种时死亡3只,存活的7只在(18.0±2.0)d时,处于濒死状态;Micro CT检查,肺转移率为5/7,肺转移灶直径1.5～3.5 mm;病理检查证实为绒癌.原代培养的细胞行人绒癌标志物β-hOG染色,呈阳性表达;原代培养的细胞染色体核型分析为单体、三体和多体人源性染色体,细胞染色体数目19～128条,染色体众数为70～79条,符合恶性肿瘤的遗传学特征.结论 通过尾静脉注射JEG-3细胞可以成功建立人绒癌SCID小鼠肺转移模型,5×106个细胞/ml×0.2 ml为建立模型的较为合适的细胞浓度及容积.     

顺铂耐药卵巢癌细胞鼠肾包膜下动物模型的建立及其临床意义

目的 :建立顺铂耐药卵巢癌细胞鼠肾包膜下动物模型 ,探讨其临床意义。方法 :采取肾包膜下纤维蛋白细胞凝块移植法 ,用环磷酰胺免疫抑制小鼠建立顺铂耐药卵巢癌模型。结果 :移植后 7d肿瘤体积明显增大 ,顺铂耐受细胞组与非耐受细胞组体积无明显差异 (P >0 .0 5) ;移植后细胞形态学特征无改变 ;在用顺铂治疗后 ,耐药细胞组肿瘤体积较非耐药细胞组体积大 (P <0 .0 5)。结论 :该动物模型可作为短期体内实验模型 ,能保持细胞的耐药性     

兔移植性子宫内膜肿瘤模型的建立及其生物学特性观察

目的：建立子宫内膜VX2肿瘤模型并观察其生物学特性。方法：采用子宫内膜分离组织包埋法在44只兔的子宫内膜制备VX2肿瘤模型,24只在1～6周处死,观察其生长及转移特性,20只用于观察模型自然生存期。结果：肿瘤原位移植成功率为100％;1周内肿瘤局限于子宫内膜,1～2周侵犯肌层并发生淋巴转移,2～3周浸润子宫浆膜并向同侧宫角蔓延,3～4周发生宫颈转移,4～5周发生肺转移;荷瘤兔的自然生存时间为50．6±8．1d,其转移灶与原发灶组织学类型一致,均为鳞癌。结论：此模型移植成功率高,生物学特性稳定,其转移模式与人子宫内膜肿瘤转移方式类似,为子宫内膜肿瘤的实验研究提供了较理想的大动物模型。     

人子宫内膜癌裸鼠原位移植模型的建立及生物学观察

目的 :建立人子宫内膜癌的原位移植模型 ,观察其生物学特性。方法 :利用子宫内膜癌细胞株AN3CN制备裸鼠皮下移植瘤模型 ,然后 ,将瘤组织条植入裸鼠子宫腔内 ,建立人子宫内膜癌的原位移植模型 ,用组织病理学检查、电镜观察和流式细胞仪DNA含量及染色体核型分析鉴定模型。结果 :原位移植模型的瘤细胞与AN3CN在形态和结构上一致。结论 :小鼠肿瘤的生长和转移完全模拟人子宫内膜癌的临床过程 ,为研究子宫内膜癌转移的理论和药物治疗提供了较客观的模型     

人侵蚀性葡萄胎裸鼠皮下移植瘤模型的建立及生物学特性的研究

目的 :建立人侵蚀性葡萄胎裸鼠皮下移植瘤模型 ,探讨其生物学特性。方法 :将人侵蚀性葡萄胎的新鲜组织块种植于裸鼠背部皮下 ,在无特殊病原菌 (SPF)条件下饲养裸鼠 ,原代生长后鼠间传代 ,观察移植瘤的生长特点 ,观察其组织形态学和超微结构 ,检测血中人绒毛膜促性腺激素 ( β -hCG)水平 ,利用流式细胞仪 (FCM)测定DNA含量 ,分析染色体核型。结果 :经 19个月建成人侵蚀性葡萄胎裸鼠皮下移植模型HIM - 980 1,瘤株生长稳定 ,已传代 54代。自第 2代起 ,移植成瘤率均为 10 0 % ,移植瘤潜伏期及传代间期短 ,生长速率快 ,向周围组织浸润 ,光镜和电镜下的形态学特征与患者肿瘤组织一致 ,DNA检测和染色体核型分析为非整倍体。结论 :HIM -980 1是首次建立成功的人侵蚀性葡萄胎裸鼠皮下移植瘤模型 ,为深入研究此病提供了理想的材料和动物体内瘤株。     

血管内皮生长因子C及其受体3mRNA在上皮性卵巢癌组织的表达及意义

目的 探讨血管内皮生长因子C（VEGF-C）及其受体3（VEGFR-3）mRNA在上皮性卵巢癌组织的表达及与癌周淋巴管生成及淋巴转移的关系。方法 2003-04—2004-06第三军医大学附属西南医院采用RT-PCR方法，检测VEGF-C及VEGFR-3mRNA在良、恶性上皮性卵巢肿瘤组织的表达及组织化学淋巴管染色并计数，分析其与VEGF-C mRNA表达及淋巴转移的关系。结果 VEGF-C mRNA和VEGFR-3 mRNA在卵巢良、恶性肿瘤的表达差异有显著性。VEGF-C mRNA表达阳性及有淋巴结转移组的癌组织淋巴管密度分别大于VEGF-CmRNA表达阴性及无淋巴结转移组的癌组织，二者差异均有显著性。结论 VEGF-CmRNA在上皮性卵巢癌组织的表达上调导致了癌周淋巴管生成，促进了肿瘤的淋巴道转移。     

Preoperative 18F-FDG PET/CT in the management of advanced epithelial ovarian cancer

Objective

The introduction of 18-FDG-PET/CT during preoperative evaluation of patients with epithelial ovarian cancer (EOC) has led to an increase of the detection of extra-abdominal metastases. However, the clinical impact of this upstage remains unclear.

Methods

Patients with suspected advanced EOC underwent 18-FDG-PET/CT within two weeks prior to debulking surgery.

Results

Between 2006 and 2011 95 patients met the inclusion criteria. Based on the concordance or the discrepancy of clinical and PET/CT stage, patients were divided into 3 groups (A: clinical and PET III; B: clinical III and PET IV; C: clinical and PET IV). Twenty-five patients were upstaged from FIGO stage III to stage IV by PET/CT. The proportion of patients who achieved a residual tumor < 1 cm in group B and C was similar, whereas it was significantly lower compared to group A. Similarly, complete response to adjuvant chemotherapy was achieved more frequently in patients in group A. PFS was similar in the three groups (17, 17 and 12 months in group A, B and C), as well as OS (51, 41 and 35 months).

Conclusions

PET/CT is able to detect distant metastases in EOC patients. The presence of extra-abdominal disease probably indicates a more aggressive disease which also shows a lower response to standard chemotherapy. However, upstaged patients have a similar prognosis compared to stage III patients, probably because intra-abdominal disease is more likely to lead patients to death. This might also explain why residual tumor is the most important prognostic factor for advanced EOC patients.     

Aurora-A/STK15在上皮性卵巢癌组织中的表达及其与预后的关系

目的探讨Aurora-A/STK15在上皮性卵巢癌组织中的表达变化及其与预后的关系。方法应用显微切割法从石蜡切片上提取总RNA,实时定量PCR和免疫组化方法检测80例原发性卵巢上皮性癌和29例正常卵巢组织中Aurora-A/STK15mRNA及蛋白水平的表达,分析其作为卵巢癌标志物的可行性以及与预后的相关性。结果上皮性卵巢癌组织中Aurora-A/STK15mRNA的表达明显高于正常卵巢组织[（60.5±74.2）对（2.19±1.4）,P〈0.01],但mRNA的表达水平与卵巢癌的分期和分级以及生存时间无关,P〉0.05;上皮性卵巢癌Aurora-A/STK15蛋白的阳性表达率也明显高于正常卵巢组织（87.3%对6.9%,P〈0.05）,蛋白表达水平随病理分级和手术分期增加而增加,与病理分级密切相关（P〈0.05）,而与手术分期的相关性无统计学意义（P〉0.05）;在行理想肿瘤细胞减灭术并行术后辅助泰素化疗的病例组,Aurora-A/STK15蛋白高表达预后好,总的生存时间较低表达病例生存时间长（P〈0.05）;在除泰素以外的铂类药物化疗组,Aurora-A/STK15蛋白高表达则与不良预后相关,高表达者总的生存时间短（P〈0.05）。结论Aurora-A/STK15在上皮性卵巢癌中存在异常表达,且与卵巢癌的分化程度以及手术联合辅助化疗的预后密切相关,有望成为卵巢癌个体化治疗疗效的预测因子。     

Age contrasts in clinical characteristics and pattern of care in patients with epithelial ovarian cancer

OBJECTIVE: The objective of this study was to document and highlight aspects of ovarian cancer treatment that pertain, especially, to elderly women. METHODS: Data were collected retrospectively from all epithelial ovarian cancer patients who were diagnosed in the Gynecologic Oncology Unit at Meir Hospital, Kfar-Saba, Israel, from January 1994 to December 1998. RESULTS: The study group comprised 143 patients (<70 years n = 97, > or =70 years n = 46). Both groups presented with the same distribution of stages. The elderly group had fewer primary debulking surgical interventions (54.3%) than the younger group (84.5%) (P = 0.001)). Age was not a limiting factor in achieving optimal debulking in those patients who did undergo surgery (older 53%, younger 54%). Almost 92% of the younger patients entered a first-line chemotherapy regimen compared to 65.2% of the older patients (P = 0.001). The elderly patients were more likely to receive neoadjuvant chemotherapy (43.3.3% vs 13.4%, P < 0.01) and hematological toxicity was significantly more common (75% vs 36.3%; P = 0.001), although no significant difference was noted between the groups in Grade 3-4 patients (> or =70 years, 62.5% vs <70 years, 45.5%; P = 0.2). The elderly patients were more likely to have dose reductions and treatment delays compared to the younger patients (60% vs 22.4%; P < 0.001, and 46.6% vs 19.1%; P = 0.004, respectively) and they had similar overall response rates (RR) and complete response (80% vs 87.6% and 60% vs 71.9%, respectively). CONCLUSIONS: Older women who present with the same distribution of stages as their younger counterparts are likely to be treated more conservatively than younger ovarian cancer patients. In this study, however, when surgery was performed, the optimal tumor debulking rates were similar in each group. Although high morbidity, most often hematological toxicity, occurs in elderly patients following chemotherapy, the overall RR compared favorably with that of younger patients.     

Aberration of the PI3K/AKT/mTOR signaling in epithelial ovarian cancer and its implication in cisplatin-based chemotherapy

Objective

This study was to investigate the role of the PI3K/AKT/mTOR signaling in epithelial ovarian cancer development and its mechanism in cisplatin-based chemotherapy.

Study design

Western blot and RT-PCR were used to determine the expression of PI3K-p85 subunit at protein and mRNA levels in normal and cancerous ovarian epithelium. SKOV3/DDP cells and SKOV3/MCA (multicellular aggregates) were constructed as chemo-resistant models. The role and mechanism of AKT specific inhibitor or shRNA in different models before and after cisplatin treatment were determined by multiple cellular and molecular approaches such as cell growth assay, flow cytometry, and western blot.

Results

PI3K-p85 subunit was detected in 33 out of 39 epithelial ovarian cancer specimens at protein level, but not detected in normal ovarian epithelium. A significant over-expression of PI3K-p85 subunit at mRNA level was observed in tumor tissues, and an increasing trend in advanced stage was also observed. Elevated activation of the AKT/mTOR/Survivin signaling was detected in SKOV3/DDP cells and SKOV3/MCA. Down-regulation of AKT by triciribine or shRNA transfection could attenuate cisplatin resistance through mTOR/Survivin signaling.

Conclusions

The PI3K/AKT/mTOR signaling was involved in epithelial ovarian cancer development and cisplatin-based chemotherapy, and down-regulation of AKT could be an effective adjuvant antitumor therapy.     

Expression of the c-Met in advanced epithelial ovarian cancer and its prognostic significance

The purpose of this study was to evaluate the prognostic significance of c-Met expression in advanced cases of epithelial ovarian carcinoma. Paraffin-embedded tissues from 41 stage IIIC primary ovarian adenocarcinoma were stained immunohistochemically for c-Met expression. The expression of c-Met was correlated with conventional clinicopathologic parameters and with overall survival of the patients. c-Met expression was found in 60.9% of cases. This clinicopathologic study showed that epithelial ovarian carcinomas with c-Met expression had higher histologic tumor grade and were more frequently associated with para-aortic lymph node metastasis (P < 0.05). In multivariate analysis, c-Met expression remained as a statistically significant predictor for survival with histologic grade. The patients with stage IIIC epithelial ovarian cancers whose tumors expressed c-Met were more likely to have high-grade tumors, have more para-aortic lymph node involvement, and have a significantly worse overall survival than those whose tumors were c-Met negative. In conclusion, c-Met expression might be a potential prognostic marker for patients with advanced-stage epithelial ovarian cancers.     

The epidemiology of epithelial ovarian cancer: a review

Banks E, Beral V, Reeves G. The epidemiology of epithelial ovarian cancer: a review. Int J Gynecol Cancer 1997; 7: 425–438.
This paper presents a review of the scientific literature investigating the epidemiology of epithelial ovarian cancer. Epithelial ovarian cancer is an important cause of cancer death among women in industrialized countries, and incidence increases with age. It is more common among women of low parity and among those with a family history of the disease. Oral contraceptive use, hysterectomy, oophorectomy, and sterilization have all been shown to protect against epithelial ovarian cancer, while the role of other reproductive and environmental factors, such as infertility, fertility drugs and hormone replacement therapy, is less clear. The evidence to date is discussed in this review.     

TrkB及其配体BDNF在卵巢上皮癌中的表达及其临床意义

目的检测酪氨酸激酶B（tyrosine kinase,TrkB）及其配体脑源性神经营养因子（brainderived neurotrophic factor,BDNF）在卵巢上皮癌（EOC）中的表达,并探讨其临床意义。方法选择中山大学附属肿瘤医院病理存档的石蜡标本108例,其中10例正常卵巢、17例良性卵巢肿瘤、21例卵巢交界性肿瘤和60例卵巢上皮癌,采用免疫组化链霉素抗生物素蛋白-过氧化物酶法（SP法）检测TrkB及其配体BDNF蛋白的表达水平,分析其与临床病理因素之间的关系。结果 TrkB和BDNF蛋白在正常卵巢组织和良性卵巢上皮性肿瘤中无表达;在交界性和卵巢上皮性癌中的阳性表达率分别为19.0%、71.7%和14.0%、51.7%（P〈0.05）。TrkB和BDNF蛋白在EOC组中的表达水平与其FIGO分期及病理分级有关,在Ⅲ期＋Ⅳ期、低分化组（G3）比Ⅰ期＋Ⅱ期、高中分化（G1、G2）组表达率高（P〈0.05）。TrkB和BDNF在卵巢上皮癌中的表达水平呈正相关（r=0.428,P〈0.05）。结论卵巢上皮癌中存在TrkB和BDNF的异常高表达。TrkB和BDNF可能协同作用促进了卵巢上皮癌的发生、发展,并有望成为卵巢上皮癌基因治疗的新靶点。     

Ifosfamide/mesna plus adriamycin as salvage therapy of advanced epithelial ovarian cancer

Linasmita V, Wilailak S, Srisupundit S, Tangtrakul S, Bullangpoti S,Israngura N. Ifosfamide/mesna plus adriamycin as salvage therapy of advancedepithelial ovarian cancer. Int J Gynecol Cancer 1997; 7:388–391.
The objective of this study was to determine the efficacy and toxicity ofifosfamide/mesna plus adriamycin as salvage therapy for epithelial ovariancancer (EOC). From October1992 to September 1995, patients with advanced EOC who had their recurrentdisease after platinum and cyclophosphamide received ifosfamide(IFX)/mesna andadriamycin (ADR). IFX wasadministered as a 24-hour infusion at a dose of 1.2 g/m² daily forthree days and 1.2 g/m² of mesna were added to the infusion solutionand run for the subsequent 24 hours after completion of the IFX. ADR was given as a singleintravenous bolus at a dose of 50 mg/m² on day 1. Measurementsinclude objective response,duration of response and toxicities to the therapy regimen. There were 20patients who received the therapy regimen, a total of 54 cycles: their medianage was 50 years, range25–71. Of the 17 evaluable patients, two achieved clinical response (1CR,1PR) for an objective response rate of 11.8%. However, among 14 patientswith platinum-refractorydisease, the response rate was only 7.1% (95% CI 0.2%,33.9%) (1CR, 0PR). At the time of the report, the patient is alivewithout evidence of disease. Eightpatients (47.0%) had stable disease. Median time to progression was fivemonths, range two to six. Alopecia and microscopic hematuria were the commontoxicities. In conclusion,the combination of ifosfamide/mesna and adriamycin has tolerable toxicities,but its efficacy as salvage therapy in platinum-refractory epithelial ovariancancer is modest.