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1.
目的研究探讨葡萄膜黑色素瘤体外化疗对6种药物的敏感性及其影响机制。方法葡萄膜黑色素瘤体外原代与传代培养,抗HMB45免疫组织化学估计肿瘤细胞纯度,MTT法检测对5-氟脲嘧啶、顺铂、噻替哌、阿霉素、长春新碱和足叶乙甙6种化疗药物的敏感性,蛋白印迹法检测耐药基因bcl-2和LRP的表达,分析其表达程度与耐药性的关系。结果体外药敏试验显示6种化疗药物临床常规剂量,对肿瘤细胞的抑制率均不能达到50%,蛋白印迹结果显示bcl-2在体外培养的肿瘤细胞中均有较高表达,而LRP表达在不同类型的肿瘤细胞中不同,表达的高低与耐药程度呈正相关。结论葡萄膜黑色素瘤体外对临床常用6种化疗药物具有较高耐受性,LRP和bcl-2均与其天然耐药性有关,LRP可能起主要作用。 相似文献
3.
目的:探讨蛋白酶活化受体2(protease-activated receptor 2,PAR2)在葡萄膜黑色素瘤(uveal melanoma,UM)中的表达,以及沉默人UM 细胞系M23中PAR2基因对细胞增殖和侵袭的影响。 方法:选取2012-02/2017-12在我院行手术治疗且资料完整的UM患者45例45眼,选取同期因眼部外伤行眼球摘除且葡萄膜正常的患者30例30眼,实时荧光定量PCR术检测UM和正常脉络膜组织中PAR2基因表达,培养M23细胞并分为PAR2干扰组、阴性对照序列组和空白组,实时荧光定量PCR技术检测细胞中PAR2基因表达,MTT法检测细胞增殖能力,Transwell法检测细胞迁移和侵袭能力。 结果:UM组织中PAR2 mRNA相对表达量为1.73±0.13,正常脉络膜组织中PAR2 mRNA相对表达量为1.06±0.10,差异有统计学意义(t=23.732,P<0.01); UM组织中PAR2 mRNA相对表达量与病理学类型、巩膜浸润、视盘受累和眼外生长有关,差异有统计学意义(P<0.05); PAR2干扰组细胞中PAR2 mRNA相对表达量低于阴性对照序列和空白组,差异有统计学意义(P<0.05); PAR2干扰组细胞24、48、72和96h时吸光度A值低于阴性对照组和空白组,差异有统计学意义(P<0.05); PAR2干扰组迁移细胞数和侵袭细胞数低于阴性对照序列组和空白组(P<0.05)。 结论:PAR2在UM组织中呈高表达,且与肿瘤高转移风险有关,特异性沉默M23细胞中PAR2基因表达可有效抑制细胞增殖、迁移及侵袭。 相似文献
4.
Case report: We report a case of choroidal melanoma metastatic to the liver diagnosed by fine-needle aspiration.The biopsy sample was immunostained for COX-2 and c-kit. Comments: Accurate diagnosis and identification of potential therapeutic targets are important for subsequent therapy and can be achieved by radiologically guided fine-needle aspiration biopsy. 相似文献
5.
AIMS: Overexpression of c-myc protein has independent prognostic significance in a variety of primary and metastatic cutaneous melanomas which suggests a possible role for this gene in melanomagenesis. We have therefore examined the importance of this oncogene in uveal melanoma and studied the coexpression of two other gene products, Bcl-2 and p53, which might contribute to its effect. METHODS: The percentage of cells positive for nuclear c-myc expression was estimated by flow cytometric analysis of nuclei extracted from paraffin blocks. The expression of Bcl-2 and p53 protein was assessed by immunohistochemistry. A total of 71 tumours were studied and the results compared with survival with a mean follow up period of 6 years. RESULTS: c-myc was expressed in > 50% of the cells by 70% of the tumours, and was independently associated with improved survival in a Cox multiple regression-model. Although Bcl-2 was expressed by the majority of the cells in 67% tumours, it was without effect on prognosis. None of the cases studied showed convincing positivity for p53. Analysis of coexpression showed that the best survival was seen in c-myc+/Bcl-2+ tumours and the worst in c-myc-/Bcl-2-tumours. CONCLUSION: The finding of improved rather than reduced survival in c-myc positive tumours is at variance with skin melanoma. There was no evidence to suggest that c-myc was modulated by upregulation of Bcl-2 or p53 inactivation/mutation. Although Bcl-2 is unlikely to have any effect on tumour growth or metastasis, it could contribute to the general lack of susceptibility to apoptosis in these tumours. 相似文献
7.
AIMS: To investigate the current referral pattern and delays in treatment of patients with primary uveal melanoma. METHODS: 184 consecutive Finnish patients with uveal melanoma diagnosed between July 1994 and June 1999 were eligible, and 159 were enrolled (inclusion rate, 86%). Their mean age was 60 years (range 14-87). The dates of visits to dispensing optician, physician, ophthalmologist and ocular oncologist, the presence of symptoms, and reason for consultation were determined by structured telephone interview. Time intervals to treatment planning and treatment were calculated. RESULTS: 139 patients (87%) had symptoms at presentation and 44 patients (28%) had been seen by an ophthalmologist less than 2 years previously. The median height of the tumour was 6 mm (range, 1.0-17.0) and its largest basal diameter 11 mm (range 2.5-22.0) at diagnosis. Melanoma developed from a previously detected presumed naevus in 13 patients (8%). When the first contact was a dispensing optician (15%) the median time to treatment planning was 22 days (range 1-1156). When a physician other than an ophthalmologist (19%) was contacted the delay was 68 days (range 0-1283) and when an ophthalmologist (65%) was seen it was 34 days (range 1-1426). These differences were not significant (p=0.32). The chance of being referred at first visit was 89%. Median time to treatment was not associated with symptoms (p=0.16) and tumour volume (p=0.29), but it was significantly different between patients who were and were not referred at first visit (140 days v 34 days; p<0.001) and between those treated by ruthenium and iodine brachytherapy (59 days v 33 days; p=0.009). CONCLUSIONS: Analysis of delays in management indicates that earlier treatment could be achieved if dilated fundus examinations were performed without exceptions, all suspicious naevi were referred for a second opinion, and if the patients with melanoma were referred to the ocular oncology service concurrently with staging examinations done at the regional hospital. 相似文献
8.
目的 探讨黑色素抗原转录子Brn-2在人脉络膜黑色素瘤细胞系中的表达及其对T细胞可识别抗原-1(melanoma antigen recognized by T cells 1,MART-1)启动子活性的影响。方法 采用RT-PCR和Western blot检测人脉络膜黑色素瘤细胞系92-1、92-2、Me1285和Ocm3 细胞中Brn-2的mRNA和蛋白表达水平。将含不同长度MART-1基因启动子序列和荧光素酶报告基因的表达质粒pGL3-Luc构建成融合表达载体pGL3-p286-Luc及pGL3-p2956-Luc,与pEV-Brn-2融合表达质粒共转染脉络膜黑色素瘤细胞系细胞。分别测量四种细胞系中p286组、p2956组、p286+Brn-2组、p2956+Brn-2组细胞荧光素酶表达变化,并观察Brn-2对上述细胞MART-1基因启动子活性的影响。结果 人脉络膜黑色素瘤细胞系92-1、92-2、Me1285、Ocm3细胞系均可检测到Brn-2 mRNA和蛋白的表达,Brn-2蛋白电泳带大致在相对分子质量47 000处。pGL3-p2956-Luc及pGL3-p286-Luc重组质粒经ApaI和NheI双酶切可切出2956 bp和286 bp 2个条带,pEV-Brn-2重组质粒经EcoRI和BamHI双酶切可切出1329 bp条带。pEV-Brn-2重组质粒分别对人脉络膜黑色素瘤细胞系Ocm3、Mel285、92-2、92-1进行磷酸钙法转染,Western blot检测可见四种细胞系均表达Brn-2蛋白。缺乏Brn-2时,所有细胞的MART-1的p286均显示出活性,MART-1阳性细胞系(92-1、92-2、Ocm3)p286活性高于阴性细胞系(Mel285)。p2956活性不同细胞系表现不同,92-1、92-2中较高,Ocm3中其活性与细胞密度相关,MART-1阴性细胞系中p2956近乎无活性。共转染Brn-2后明显下调92-1、92-2、Ocm3中启动子p286及p2956活性(P<0.05);阴性细胞系Mel285中,Brn-2对启动子活性无影响(P>0.05)。结论 Brn-2表达于人脉络膜黑色素瘤细胞,并下调阳性细胞系MART-1启动子活性。 相似文献
9.
目的:探讨高迁移率族蛋白A1(HMGA1)蛋白在葡萄膜黑色素瘤(UM)组织中的表达,以及抑制该基因表达对细胞增殖和侵袭的影响。 方法:选取2014-02/2019-08在我院接受手术治疗的UM患者53例53眼,同期留取因外伤摘除眼球的正常葡萄膜组织34例34眼。采用免疫组织化学法检测组织中HMGA1蛋白表达。将体外培养的人UM细胞系M23分为HMGA1下调组、阴性对照组和空白组,分别转染HMGA1干扰序列、阴性对照序列和不作任何处理,采用实时荧光定量PCR术检测HMGA1 mRNA表达,CCK-8法检测细胞增殖能力,Transwell法检测细胞迁移和侵袭能力。 结果:UM组织中HMGA1蛋白阳性表达率为77%,高于正常葡萄膜组织中的29%(P<0.001); 与未发生巩膜浸润、未累及睫状体和未发生眼外生长相比,HMGA1蛋白在发生巩膜浸润、累及睫状体和发生眼外生长的组织中阳性表达率升高(均P<0.05)。与阴性对照组和空白组相比,HMGA1 mRNA在HMGA1下调组细胞中相对表达量降低,且HMGA1下调组细胞培养24、48、72、96h时吸光度OD值降低,迁移细胞数和侵袭细胞数均明显减少(均P<0.05)。 结论:UM组织中HMGA1蛋白阳性表达率升高,下调M23细胞中HMGA1表达可减少细胞增殖,抑制细胞迁移和侵袭。 相似文献
10.
Uveal melanomas are the commonest ocular tumour of adults and are characterized by reproducible alterations of chromosomes 1, 3, 6 and 8. These alterations are of prognostic relevance and have also be shown to correlate to high risk and low risk metastatic categories of uveal melanoma as defined by micro-array analysis. It is, however, possible that a catalogue of relevant genetic alterations, involving gene rearrangement rather than amplification, have as yet eluded identification. To address this point we examined 14 primary uveal melanomas, using 24 colour multiplex fluorescence in situ hybridization (M-FISH). All tumours were karyotyped following G-Banding, and M-FISH was performed to confirm and clarify the identity of abnormal chromosomes. M-FISH data were obtained from all tumours and was able to establish the nature of most abnormalities not fully characterized by cytogenetics. Abnormalities of chromosome 6 were far more frequent than previously indicated, in approximately 70% of cases, indicating they have been substantially underrepresented in past studies of uveal melanoma. Spindle melanomas were found to have novel rearrangements affecting in particular chromosomes 6, 15 and 18, suggesting that juxtaposition of genes through translocational events may play a role in the development of some uveal melanomas. In conclusion, this study is the largest of primary uveal melanoma analysed by M-FISH and indicates that alterations of chromosome 6 have previously been underestimated. Furthermore spindle melanomas are prone to rearrangements affecting chromosomes 6, 15 and 18, which may relate to early changes in uveal melanoma development or associate with those melanomas of a more differentiated status. 相似文献
11.
目的 研究LncRNA MT1JP对葡萄膜黑色素瘤细胞迁移和侵袭的影响及机制.方法 实时荧光定量PCR测定Ln-cRNA MT1JP在葡萄膜黑色素瘤细胞系SP6.5、M23及正常视网膜色素上皮细胞系ARPE-19中的表达,将SP6.5细胞分成3组,即MT1JP组、siMT1JP组和NC组,分别转染pcDNA3.1-MT1JP、pcDNA3.1-siMT1JP及pcDNA3.1空载体,采用细胞划痕实验和Transwell实验测定3组划痕愈合率和侵袭细胞数,Western blot测定P53蛋白表达水平.结果 葡萄膜黑色素瘤细胞系SP6.5和M23中LncRNA MT1JP的相对表达量分别为0.20±0.02和0.31±0.01,均显著低于正常视网膜色素上皮细胞系ARPE-19的1.0(均为P<0.01).siMT1JP组划痕愈合率为61.5%±3.7%,MT1JP组为10.6%±1.7%,NC组为20.0%±2.1%,3组间两两相比差异均有统计学意义(均为P<0.01).在200倍视野下,siMT1JP组侵袭细胞数为(75.6±6.8)个,MT1JP组侵袭细胞数为(10.3±2.6)个,NC组为(29.50±3.9)个,3组间两两相比差异均有统计学意义(均为P<0.01).siMT1JP组P53蛋白相对表达量为0.41±0.04,显著低于NC组的1.0(P <0.01);MT1JP组P53蛋白相对表达量为5.73±0.62,显著高于NC组的1.0(P<0.01).结论 LncRNA MT1JP抑制葡萄膜黑色素瘤细胞的转移和侵袭,其机制可能与上调P53蛋白表达有关. 相似文献
12.
AIM: To detect how BRCA-associated protein 1 (BAP1) regulates cell migration in uveal melanoma (UM) cells.
METHODS: Wound healing and transwell assays were performed to detect UM cell migration abilities. Protein chip, immunoprecipitations and surface plasmon resonance analyses were applied to identify BAP1 protein partners. Western blot and calpain activity assays were used to test the expression and function of calpastatin (CAST).
RESULTS: CAST protein was confirmed as a new BAP1 protein partner, and loss of BAP1 reduced the expression and function of CAST in UM cells. The overexpression of CAST rescued the cell migration phenotype caused by BAP1 loss.
CONCLUSION: BAP1 interacts with CAST in UM cells, and CAST and its subsequent calpain pathway may mediate BAP1-related cell migration regulation. 相似文献
13.
Purpose: Our objective was to introduce immunomagnetic separation (IMS) in ocular research by evaluating the possibility of detecting tumour cells in bone marrow (BM) and peripheral blood (PB) samples and validating the captured cells as melanocytic cells. Methods: Mononuclear cell (MNC) fractions isolated from BM and PB in uveal melanoma patients were examined for tumour cells using our IMS method. Sheep‐anti‐mouse IgG antibody‐coated super paramagnetic particles were conjugated to an anti‐melanoma antibody. Microscopy of the magnetic fraction isolated from MNCs was performed to identify and count the number of bead‐rosetted cells. The finding of at least two rosettes with coated beads in a 20‐μl fraction of a sample was registered as a positive test. The melanocytic nature of the tumour cells was ascertained with a double labelling procedure using fluorescent microparticles. Results: Using IMS in a study of 328 patients, tumour cells were at initial diagnosis found in BM and PB in 29.9% and 1.6% of cases, respectively. In positive samples, a median of 56 tumour cells (range 2–500) were identified. The captured cells were documented to be of melanocytic origin by the simultaneous binding of fluorescent beads coated with another melanoma‐associated antibody. Conclusions: The IMS method was sensitive and efficient in the detection of occult melanoma tumour cells in BM. The validity of the immunomagnetic technique was strengthened by verifying the melanocytic characteristics of the isolated cells. The IMS procedure identifies intact, vital tumour cells, permitting further molecular characterization, an advantage which makes this method attractive for extended use. The clinical relevance of the findings will be further investigated in follow‐up studies with repeated sampling and characterization of the isolated tumour cells. 相似文献
14.
Purpose: To determine and compare cyclo‐oxygenase‐2 (COX‐2) expression in photon‐radiated and non‐radiated malignant uveal melanomas and to analyse the correlation between COX‐2 expression and prognosis. Methods: Immunohistochemical staining for COX‐2 was performed on 21 uveal melanomas that were endoresected after prior stereotactic radiotherapy with photons and on 22 tumours that were treated by endoresection without prior radiotherapy. COX‐2 staining was further analysed in respect to cell type, maximal prominence, time interval between radiotherapy and surgery, apoptotic index (AI), proliferative index (PI) and the development of metastatic disease. Results: There was no difference in COX‐2 expression between radiated and non‐radiated melanomas ( P > 0.15). COX‐2 staining correlated with neither the tumour prominence ( P > 0.40) nor the AI or the PI (both P > 0.35). Tumours with high COX‐2 expression were significantly more likely to develop metastasis ( P = 0.022). Conclusion: Radiotherapy with photons does not induce COX‐2 expression in malignant melanomas of the uvea. But high COX‐2 expression may be a marker for poor prognosis. 相似文献
15.
目的探讨IL-1β、IL-6反义寡核苷酸脂质体抑制后发性白内障的可行性和有效性。方法将20只健康新西兰大白兔双眼行透明晶状体囊外摘出术,均设左眼为实验眼,术后前房立即注射IL-1β、IL-6反义寡核苷酸脂质体,右眼为对照眼,术后前房注射单纯脂质体。分别于术前1d、术后1d、3d、1周、2周、1月、2月和3月抽取房水,用ELISA双抗体免疫夹心法检测IL-1β、IL-6的含量,定期裂隙灯检眼镜观察后囊混浊情况,术后3月行组织病理学检查。结果(1)术后3月实验眼与对照眼发生晶状体后囊膜混浊的眼数分别为16眼及20眼,差异有极显著意义(P=0.002<0.01);(2)实验眼和对照眼房水中IL-1β的浓度均值分别为(8.570±1.686)×10-9g·L-1、(11.900±3.873)×10-9g·L-1,有显著性差异(P=0.002<0.01),IL-6的浓度均值分别为(12.530±4.068)×10-9g·L-1、(16·600±6·636)×10-9g·L-1,亦有显著性差异(P=0.033<0.05);(3)光镜和电镜下实验眼晶状体后囊膜细胞增生不活跃,未发现眼内毒性反应。结论前房注射IL-1β、IL-6反义寡核苷酸脂质体对后发性白内障可能有一定的抑制作用。 相似文献
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目的:基于癌症基因组图谱(TCGA)数据库分析SLC52A2与葡萄膜黑色素瘤(UM)的相关性,初步探讨SLC52A2对UM患者预后的影响及可能的机制。方法:通过TCGA数据库下载收集80例UM患者的临床信息资料和SLC52A2的mRNA表达数据,根据SLC52A2表达量采用中位数法将80例患者分为SLC52A2高、低表达组,分析SLC52A2的表达水平与患者临床病理特征及预后的关系。对患者的年龄、性别、临床分期、病理分期、SLC52A2的mRNA表达进行Cox单因素和多因素分析,寻找UM预后因子。GSEA富集分析预测SLC52A2在UM中可能的调控通路。结果:SLC52A2低表达患者生存预后优于SLC52A2高表达患者(P<0.05)。SLC52A2高、低表达组患者的年龄、性别、临床分期、病理分期均无差异(P>0.05)。Cox多因素分析表明,SLC52A2高表达是UM患者预后的危险因素。结合SLC52A2表达和临床病理学特征开发的列线图预测模型,可较为准确地预测UM患者的生存概率。SLC52A2高、低表达组Th2细胞、Treg细胞的浸润丰度分别比较有差异(均P<0.... 相似文献
17.
目的:分析葡萄膜恶性黑色素瘤患者血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)的表达水平是否与葡萄膜恶性黑色素瘤的转移和预后有关。 方法:采用ELISA法检测葡萄膜恶性黑色素瘤患者和正常对照组中外周血VEGF水平,并采用RT-PCR和Western-blot检测葡萄膜恶性黑色素瘤患者肿瘤组织中VEGF mRNA和VEGF蛋白表达水平。对葡萄膜恶性黑色素瘤患者进行随访,用Kaplan-Meier法分析基线VEGF水平与患者术后转移的关系。 结果:葡萄膜恶性黑色素瘤患者外周血VEGF水平较健康者升高。就诊时肿瘤发生转移的患者VEGF mRNA和VEGF蛋白表达水平均较未发生转移的患者升高。在随访过程中,血清VEGF≥700pg/mL的患者,肿瘤发生转移的风险较高。 结论:葡萄膜恶性黑色素瘤患者中VEGF的表达水平与葡萄膜恶性黑色素瘤的转移密切相关。 相似文献
18.
目的 探讨E2 6转录因子 1(E2 6transformation specific 1,Ets 1)在不同类型的葡萄膜黑色素瘤中的表达及其与预后的相关关系。方法 以原位杂交和免疫组织化学法检测 78例葡萄膜黑色素瘤中Ets 1mRNA和蛋白的表达 ,并按WHO 1980年的标准进行分型 :梭型细胞型、类上皮细胞型和混合细胞型。结果 78例中 ,梭型细胞型占 2 1例 ,类上皮细胞型占 34例 ,混合细胞型占 2 3例。Ets 1mRNA和蛋白在 3种类型的葡萄膜黑色素瘤中均有表达 ,但表达强度随梭型细胞型 ,混合细胞型和类上皮细胞型依次递增。回访37例患者 ,其中梭型细胞型 18例 ,平均生存时间为 (78.33± 2 4 .6 9)月 ;混合细胞型 10例 ,平均生存时间 (6 1.4 4± 2 0 .4 6 )月 ;类上皮细胞型 9例 ,平均生存时间 (36 .76± 12 .19)月。患者生存时间与Ets 1表达强度呈负相关。结论 Ets 1可能在葡萄膜黑色素瘤的转移、浸润中发挥重要作用 ,Ets 1的检测可作为葡萄膜黑色素瘤恶性程度的参考指标。 相似文献
19.
Uveal melanoma (UM) is the most common intraocular malignancy. Approximately 50% of UM patients die of metastases, which mainly arise from primary tumors with loss of an entire chromosome 3 (monosomy 3). To identify cell lines with monosomy 3 that may serve as a model system for UM with high metastatic potential, we determined the chromosome 3 status of previously established and frequently used UM cell lines by microsatellite analysis (Mel202, Mel285, Mel290, 92-1, OMM-1, OCM-1, OCM-3, OCM-8) and cytogenetic analysis (Mel202, Mel285, OCM-8). We found that none of these cell lines has monosomy 3. Therefore we established and characterized two novel cell lines, UPMM-1 and UPMM-2 that are both developed from primary uveal melanoma tissue samples with monosomy 3. The cell line UPMM-1 has retained the chromosome 3 status of the primary tumor. In UPMM-2 chromosome 3 has undergone duplication (isodisomy) and is present on the background of a hypotetraploid karyotype. Our data suggest that, UPMM-1 may serve as a model system to study the mechanisms underlying the metastatic potential of uveal melanomas with monosomy 3. 相似文献
20.
目的 探讨石蒜碱对葡萄膜黑色素瘤细胞的作用,并分析其可能机制。方法 取B16F10葡萄膜黑色素瘤细胞株,按石蒜碱浓度为0μmol·L -1(对照组)、1.560μmol·L -1、3.125μmol·L -1、6.250μmol·L -1、12.500μmol·L -1、25.000μmol·L -1进行分组培养,24 h后,CCK-8法检测各组B16F10细胞的存活率;RT-PCR分析各组B16F10细胞VEGF-A mRNA的表达,ELISA检测各组B16F10细胞VEGF-A蛋白表达;流式细胞仪检测各组B16F10细胞的细胞周期及凋亡情况。结果 与对照组相比,1.560μmol·L -1、3.125μmol·L -1、6.250μmol·L -1、12.500μmol·L -1、25.000μmol·L -1石蒜碱作用于B16F10细胞24 h后,能够呈浓度依赖性... 相似文献
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