粒细胞巨噬细胞集落刺激因子对人卵巢细胞株生长的影响

目的Ｌ：了解粒细胞巨噬细胞集落刺激因子（ＧＭ－ＣＳＦ）对人卵巢癌细胞株生长的影响。方法：采用ＸＴＴ比色法及流式细胞仪检测ＧＭ－ＣＳＦ对卵巢癌细胞株ＳＫＯＶ３、３ＡＯ及ＣＡＯＶ３细胞生长的影响。结果：用ＸＴＴ法检测ＧＭ－ＣＳＦ对ＳＫＯＶ３、３ＡＯ及ＣＡＯＶ３细胞的生长均无明显影响;流式细胞仪检测也未发现ＧＭ－ＣＳＦ对ＳＫＯＶ３、３ＡＯ及ＣＡＯＶ３的增殖指数有明显影响。结论：在卵巢癌化疗过程中应用ＧＭ     

粒细胞巨噬细胞集落刺激因子对人卵巢癌细胞株生长的影响

目的：了解粒细胞巨噬细胞集落刺激因子（ＧＭ ＣＳＦ）对人卵巢癌细胞株生长的影响。方法：采用ＸＴＴ比色法及流式细胞仪检测ＧＭ ＣＳＦ对卵巢癌细胞株ＳＫＯＶ３、３ＡＯ及ＣＡＯＶ３ 细胞生长的影响。结果：用ＸＴＴ法检测ＧＭ ＣＳＦ对ＳＫＯＶ３、３ＡＯ 及ＣＡＯＶ３ 细胞的生长均无明显影响;流式细胞仪检测也未发现ＧＭ ＣＳＦ对ＳＫＯＶ３、３ＡＯ及ＣＡＯＶ３细胞的增殖指数有明显影响。结论：在卵巢癌化疗过程中应用ＧＭ ＣＳＦ是比较安全的,对其潜在的促进肿瘤细胞增殖的顾虑也许是不必要的。     

卵巢癌抗独特型抗体6B11scFv和人粒细胞巨噬细胞集落刺激因 …

目的 构建和表达模拟人卵巢癌抗原的抗独特型单链抗体６ＢｌｌｓｃＦｖ和人粒细胞－巨噬细胞集刺激因子的融合蛋白,以提高抗独特型抗体的免疫原性,为进一步用于卵巢癌的免疫治疗提供基础。方法 采用基因工程技术,用编码连接肽的ＤＮＡ连接６ＢｌｌｓｃＦｖ和ｈＧＭ－ＣＳＦ ｃＤＮＡ,并对大肠杆菌用温度进行诱导表达;     

巨噬细胞集落刺激因子对卵泡颗粒黄体细胞性激素分泌的影响及其受体的测定

目的　了解人卵泡颗粒黄体细胞中巨噬细胞集落刺激因子 (M CSF)受体的表达 ,M CSF在体外对人卵泡颗粒黄体细胞产生雌、孕激素的影响。方法　采用免疫细胞化学染色法 ,测定 2 0例行卵母细胞浆内单精子注射治疗患者的卵泡颗粒黄体细胞的M CSF受体 ;采用酶免疫分析法 ,测定卵泡颗粒黄体细胞在M CSF(浓度分别为 0、10、2 5、5 0、10 0、2 5 0ng/ml)单独及与促卵泡激素 (FSH ,浓度75IU/ml)联合作用 72h后的上清液中雌二醇 (E2 )和孕酮 (P)的浓度。结果　约 80 %的颗粒黄体细胞膜上M CSF受体阳性。在无FSH存在时 ,M CSF空白对照颗粒黄体细胞培养上清液中E2 浓度为(2 185± 189)pmol/L ,P浓度为 (315 7± 4 0 1)nmol/L ;M CSF为 10～ 10 0ng/ml时 ,E2 浓度在 (2 789± 36 5 )～ (42 82± 318)pmol/L之间 ,P浓度在 (42 5 6± 5 95 )～ (7789± 82 8)nmol/L之间。在有FSH作用时 ,M CSF空白对照的E2 和P浓度分别为 (5 0 4 5± 4 86 )pmol/L和 (86 6 7± 92 3)nmol/L ;M CSF为 10～ 10 0ng/ml时 ,E2 浓度在 (6 5 6 7± 6 73)～ (8373± 935 )pmol/L之间 ,P浓度在 (10 999± 985 )～ (14 990±115 8)nmol/L之间。M CSF促颗粒黄体细胞E2 和P分泌的作用随浓度增加而增强 (P均 <0 0 5 ) ;M CSF与FSH共同作用下颗粒黄体     

妊高征患者血浆巨噬细胞集落刺激因子的测定

妊高征是严重威胁母儿安全的妊娠并发症,其病因和发病机理至今未定论。近年来研究发现,巨噬细胞集落刺激因子（ＭＣＳＦ）有调节胎盘发育和功能的作用。本研究采用酶联免疫吸附试验（ＥＬＩＳＡ）测定妊高征患者、正常晚孕妇女及健康非孕妇女血浆ＭＣＳＦ的水平,以...     

雌激素对卵巢癌细胞株体外生长的影响

目的 探讨１７－β雌二醇（Ｅ２）对卵巢癌细胞株体外生长和细胞周期的影响。方法 采用四甲基偶氮唑蓝（ＭＴＴ）比色法和流式细胞术（ＦＣＭ）的方法,观察了卵巢癌细胞株ＣＡＯＶ３和ＯＣＶＡＲ３在加入含有不同浓度１７－β Ｅ２培养液后细胞活性和细胞周期的变化;同时采用免疫组化方法,测定癌细胞在加入１７－β Ｅ２前后雌激素受体表面的影响。结果 （１）浓度为０．２５～５．００ｎｍｏｌ／Ｌ的１７－β Ｅ２,可轻度     

超促排卵周期中巨噬细胞集落刺激因子的测定

目的　探讨巨噬细胞集落刺激因子 (M CSF)对卵泡发育、排卵、卵子受精及胚胎质量的影响。方法　用酶联免疫吸附试验方法测定 3 6例超促排卵患者的卵泡早期、卵泡中期和取卵日血清中M CSF水平 ,以及取卵日卵泡液中M CSF水平。结果　超促排卵周期中 ,血清M CSF水平呈逐渐升高趋势 ,取卵日达高峰。取卵日卵泡液M CSF水平 [(12 4 5 .7± 4 3 .6)kU/L]显著高于同日血清M CSF水平 [(983 .6± 2 9.6)kU/L]。在找到卵子、受精卵及直径≥ 16mm和容积≥ 2ml的卵泡中 ,其卵泡液M CSF水平分别为 (13 2 7.3± 4 2 .8)kU/L、(13 5 6.2± 3 4.7)kU/L和 (12 97.6± 3 3 .7)kU/L ;而未找到卵子、未受精及直径 <16mm和容积 <2ml卵泡的卵泡液M CSF水平分别为 (10 86.7± 2 8.3 )kU/L、(1175 .4± 3 7.3 )kU/L和 (10 3 8.4± 2 5 .9)kU/L ,3者间比较 ,差异均有极显著性 (P <0 .0 1,P <0 .0 5 ,P<0 .0 5 )。但是 ,胚胎分级 3～ 4分的卵泡液M CSF水平 [(12 71.8± 3 7.3 )kU/L]与胚胎分级 1～ 2分的卵泡液M CSF水平 [(13 12 .6± 5 1.2 )kU/L]比较 ,差异无显著性 (P >0 .0 5 )。结论　M CSF通过参与卵泡发育、成熟而影响卵子受精 ,但不影响卵裂及胚胎质量     

糖皮质激素对人卵巢癌细胞系增殖的抑制作用

糖皮质激素对人卵巢癌细胞系增殖的抑制作用卢建张金山牟瀚舟许沈华卵巢含有性激素和糖皮质激素受体。但人们对甾体激素在卵巢癌发生、发展中的作用了解尚不多。ＨＯ－８９１０为一人卵巢癌细胞系，有研究发现，性激素对ＨＯ－８９１０细胞的增殖过程无明显作用［１］。本...     

survivin反义RNA对卵巢癌细胞株SKOV3生长的抑制作用

目的观察survivin反义RNA对卵巢癌细胞株SKOV3的生长抑制作用及其对裸鼠皮下种植瘤成瘤能力的影响。方法构建能在真核细胞中稳定表达survivin反义RNA的载体pcDNA3-SVVas,应用基因重组技术,将pcDNA3-SVVas经脂质体lipofectam ine 2000介导转染卵巢癌细胞株SKOV3(SKOV3/SVVas),同时以空载体pcDNA3转染SKOV3细胞(SKOV3/neo)作为对照;绘制细胞生长曲线,采用免疫组化、RT-PCR和流式细胞仪检测SKOV3/SVVas、SKOV3/neo和SKOV3细胞survivin mRNA及蛋白的表达和细胞凋亡率。将24只裸鼠随机分为3组,每组8只,分别将SKOV3/SVVas、SKOV3/neo、SKOV3细胞接种于裸鼠皮下,观察成瘤情况及肿瘤体积变化。结果与SKOV3细胞比较,SKOV3/SVVas细胞增殖能力明显降低;SKOV3/SVVas细胞survivin蛋白阳性细胞率及survivin mRNA表达量分别为(37.5±1.0)%、0.407±0.022,与SKOV3细胞的(81.2±0.4)%、0.793±0.042和SKOV3/neo细胞的(80.4±0.8)%、0.734±0.039比较,差异均有统计学意义(P<0.01);SKOV3/SVVas细胞凋亡率显著增加,为(27.4±9.6)%,与SKOV3和SKOV3/neo细胞比较,差异有统计学意义(P<0.05)。SKOV3/SVVas组裸鼠成瘤率降低,出现目检可见肿瘤的平均时间延长为(14.0±1.0)d,与SKOV3/neo组的(6.1±0.8)d和SKOV3组的(5.8±0.9)d比较,差异有统计学意义(P<0.01);与SKOV3、SKOV3/neo组比较,SKOV3/SVVas组裸鼠肿瘤生长速度缓慢,肿瘤体积的差异均有统计学意义(P<0.01)。结论稳定表达的survivin反义RNA能够有效抑制SKOV3细胞生长,降低survivin的表达,诱导SKOV3细胞凋亡;转染survivin反义RNA的SKOV3/SVVas细胞在裸鼠皮下的成瘤能力降低。     

CO_2对人卵巢癌细胞株生长转移相关因子表达及顺铂敏感性的影响

目的:探讨体外模拟腹腔镜二氧化碳(CO2)气腹环境对人卵巢癌细胞株SKOV3生长转移相关因子表达的影响及CO2气腹环境作用后SKOV3对顺铂(DDP)敏感性的变化。方法:将体外培养的处于对数生长期的SKOV3细胞分为4组,即CO2组、DDP组、CO2+DDP组及对照组。用实时荧光定量PCR(Real-timePCR)法及免疫细胞化学法检测各组细胞的PCNA、VEGF和CD44v6mRNA和蛋白的表达。结果:(1)CO2气体与顺铂间无交互作用(P0.05);(2)CO2处理组肿瘤细胞的PCNA、VEGF和CD44v6mRNA和蛋白表达均明显升高(P0.01);(3)DDP组与对照组、CO2+DDP组与CO2组比较,DDP处理组肿瘤细胞的PCNA、VEGF和CD44v6mRNA和蛋白表达均明显降低(P0.01,P0.01)。结论:CO2环境能促进人卵巢癌细胞株SKOV3生长转移相关因子表达;DDP对肿瘤细胞生长转移因子的表达有抑制作用;CO2环境作用后,肿瘤细胞对DDP敏感性的变化不明显。     

Renal cell carcinoma producing granulocyte colony-stimulating factor.

Severe neutrophilia caused by renal cell carcinoma secreting granulocyte colony-stimulating factor (G-CSF) is a rare manifestation of renal cancer. A 70-year-old woman presented with a 2-year history of severe anemia, severe neutrophilia and elevated serum G-CSF, which completely resolved after radical nephrectomy. Histologic study revealed a histologically high-grade, stage pT1N0M0 renal cell carcinoma. Serum G-CSF level was elevated preoperatively and returned to normal postoperatively. Immunohistochemical study of the tumor tissue using anti-G-CSF monoclonal antibody revealed positive staining in the cancer cells. Careful follow-up of white blood cell count and physical examination for neck lymph node enlargement led to the timely identification of tumor recurrence 17 months after surgery, which resulted in prompt and successful salvage immunotherapy. In this case, G-CSF appeared to contribute to the leukocytosis, as both serum G-CSF level and white blood cell count closely correlated with the clinical tumor growth. White blood cell count should be closely monitored as an indicator of disease activity in patients with G-CSF-producing renal cell carcinoma.     

Prognostic relevance of granulocyte colony-stimulating factor in ovarian carcinomas

Purpose  

Granulocyte colony-stimulating factor (G-CSF) producing tumours were found associated with poor prognosis. Unfortunately, this finding is based on several case reports only. Thus, we investigated the expression of G-CSF in the tumour cells and the tumour stroma in a large collective of patients with ovarian cancer with long-term follow-up.     

Granulocyte colony-stimulating factor on the growth of human ovarian carcinoma cells in vitro

Granulocyte colony-stimulating factor (GCSF) is a member of a family of glycoprotein hormones that stimulates the proliferation and differentiation of hematopoietic cells in vivo and in vitro. In ovarian cancer chemotherapy, GCSF is used clinically to build up bone marrow function after severe cytotoxicity to granulocytes by chemotherapy. Little is known about the effects of these cytokines on the growth of cancer cells. To study the influence of GCSF on the proliferation of ovarian cancer cells in vitro, five established ovarian cancer cell lines (OC-89-VGH, OC-117-VGH, OCPC-2-VGH, NIH-OVCAR-3, and NIH-SK-OV-3) were treated with the concentration ranging from 0.1 pg to 10 ng GCSF for 5 days and compared with vehicle control. In addition, we also examined the effect of GCSF (0. 01-1.0 ng) on eight primary cultures of fresh ovarian cancer tissues. Cell viability after treatment was measured by Cell Titer AQueous assay and expressed as a percentage of untreated control cultures. A decrease in cell growth (75-85%) was observed in OC-89-VGH, OC-117-VGH and OCPC-2-VGH cell lines while NIH-OVCAR-3 and NIH-SK-OV-3 cells showed minimal growth stimulation. However, all dose-response curves were nonsignificant, suggesting indirect effects. In the eight fresh tumor primary cultures treated with GCSF, no statistical significant difference in growth was observed. In conclusion, our data suggest that GCSF has little or no growth-modulatory effect on human ovarian carcinoma in vitro. Therefore, the clinical use of GCSF is unlikely to have a direct effect on tumor growth. Copyright Copyright 1999 S. Karger AG, Basel     

Interferon-induced changes in human ovarian carcinoma cell lines

Metcalf KS, Selby PJ, Trejdosiewicz LK, Southgate J. Interferon-inducedchanges in human ovarian carcinoma cell lines. Int J Gynecol Cancer 1997; 7 : 355–363.
Interferon α (IFNα) may have a clinical role in maintenance therapyof advanced epithelial ovarian cancer following chemotherapy. To investigatethe action of IFN, we havestudied the direct effects of IFNα and IFNγ on cell surface antigenexpression in four human ovarian cancer cell lines (Caov-3, SK-OV-3, SW626 andNIH OVCAR 3). All celllines demonstrated responses to the IFNs, indicating that IFN response elementshad been retained. The effects of IFNα and IFNγ on cell surfacephenotype were investigatedby analytical flow cytometry for a panel of 15 cell surface antigens, whichincluded tumor-associated antigens, growth factor receptors, MHC antigens,adhesion molecules andepithelial mucins. As single agents or in combination, the effects of theinterferons on cell surface phenotype were heterogeneous, with up-regulation ofMHC class I being the mostconsistent observation. IFNγ, alone or with IFNα, also up-regulatedexpression of MHC class II and ICAM-1. IFNα caused down-regulation of cellsurface expression of theCA125 antigen in NIH OVCAR 3 cells by inducing antigen shedding. This haspotential clinical implications for tumor monitoring during IFN therapy inovarian cancer.     

Characterization of two new human ovarian carcinoma cell lines

Two permanent cell lines, A7 and A10, were established from ascitic effusions obtained from patients with ovarian carcinomas. Cell growth characteristics, karyotyping, and morphological examinations by light and electron microscopy revealed the neoplastic nature of the lines. Desmosomes were found indicating an epithelial origin. Both lines showed G6PD type B. Ascites seemed to be a good source for cultivation of ovarian carcinoma cells.     

Characterization of two human ovarian carcinoma cell lines.

Two permanent cell lines, A7 and A10, were established from ascitic effusions obtained from patients with ovarian carcinomas. Cell growth characteristics, karyotyping, and morphological examinations by light and electron microscopy revealed the neoplastic nature of the lines. Desmosomes were found indicating an epithelial origin. Both lines showed G6PD type B. Ascites seemed to be a good source for cultivation of ovarian carcinoma cells.     

Nitric oxide-dependent apoptosis in ovarian carcinoma cell lines.

OBJECTIVE: In a recent study, we found different profiles of inducible nitric oxide synthase (iNOS) gene expression in the ovarian carcinoma cell lines OVCAR-3, HOC-7, and 2774 following stimulation by proinflammatory cytokines. The present study was performed to determine whether nitric oxide (NO) synthesis correlates with programmed cell death in these cells. METHODS: NO-Dependent apoptosis was detected by DNA fragmentation analysis and fluorescence-activated cell sorter analysis. RESULTS: NO formation in response to interferon gamma (IFN-gamma), interleukin-1beta (IL-1beta), and tumor necrosis factor alpha (TNF-alpha) was correlated with programmed cell death in the investigated cells. DNA fragmentation was most prominent in OVCAR-3 (34.17 +/- 1.81%), less pronounced in HOC-7 (12.86 +/- 0.45%), and undetectable in 2774 (4.54 +/- 0.40%) cells. The rate of apoptosis correlated with the amount of NO formation in cytokine-treated cells. Moreover, coincubation of OVCAR-3 and HOC-7 with the specific iNOS inhibitor aminoguanidine suppressed apoptosis induced by IFN-gamma, IL-1beta, and TNF-alpha. CONCLUSION: Our data indicate that in OVCAR-3 and HOC-7 cells, NO synthesis induced by IFN-gamma, IL-1beta, and TNF-alpha is correlated with the degree of apoptotic cell death. In clinical situations, this might in part explain the benefit of cytokine application in ovarian carcinoma patients (e.g., documented for IFN-gamma).     

The effect of amifostine on the in vitro cytotoxicity of chemotherapeutic agents in three epithelial ovarian carcinoma cell lines.

OBJECTIVES: Amifostine protects against a spectrum of toxicities induced by chemotherapy without affecting tumor cell kill. This is supported by clinical data and in vivo animal studies. However, there is a paucity of data on its effect on the tumor cytotoxicity of several chemotherapeutic agents used in recurrent epithelial ovarian cancer. This study compares in vitro cytotoxicity before and after addition of amifostine. METHODS: Three epithelial ovarian carcinoma cell lines (SKOV3, 420, 429) were exposed to cis-platinum, paclitaxel, doxorubicin, etoposide, 5-fluorouracil, bleomycin, 4-epidoxorubicin, 4-HC (activated cyclophosphamide), vincristine, actinomycin D, mitomycin C, and topotecan. Cells were pretreated with either 0 or 1.2 mM amifostine. Tumor cell kill after 6 days of incubation was measured using the ATP cell viability assay. Paired samples Student's t statistic was used to test the difference in mean ATP levels between drug-treated samples with and without pretreatment with amifostine. RESULTS: SKOV3 was sensitive to paclitaxel, actinomycin D, 4-epidoxorubicin, and vincristine. Cell line 420 was sensitive to paclitaxel, etoposide, and 5-fluorouracil. Cell line 429 was sensitive to paclitaxel and 5-fluorouracil. There was no significant difference in the mean ATP levels between drug-treated samples with and without pretreatment with amifostine for each of the sensitive drugs in all three cell lines. Similarly, there was no significant difference in the mean ATP levels in cis-platinum and 4-HC treated samples with and without pretreatment with amifostine. CONCLUSIONS: These results show that at the cellular level amifostine did not protect epithelial ovarian carcinoma cells against tumor cell kill.