C57BL/6小鼠B16黑色素瘤细胞脾转移机制

 目的　研究C57BL／6小鼠B16黑色素瘤细胞脾转移与血行转移的相关性。方法　细胞悬液接种法制备C57BL／6小鼠B16黑色素瘤细胞的荷瘤模型，HE染色法观察各组织器官中转移瘤细胞的有无及其生长状态，伊文思蓝尾静脉注射法观察蓝染部位与肿瘤转移阳性部位间的相关性。结果　接种成瘤率100 ％。12只小鼠中有9只出现脾转移，均同时伴有不同程度的局部淋巴结转移， 其中3只尚伴有肺转移。脾转移较肺转移出现早、概率高。大体观脾转移阳性部位位于脾背侧段约全长1／4区域。镜下观脾内转移瘤细胞呈散在分布，生长受抑并向成熟分化。伊文思蓝蓝染部位位于脾背侧段约全长1／4区域，与肿瘤转移阳性部位一致。结论　C57BL／6小鼠的B16黑色素瘤细胞脾转移先于其他脏器转移，其转移途径为血行转移。     

恶性黑色素瘤胃转移一例报告

恶性黑色素瘤胃转移很少见,我院发现一例,现报告如下。患者男性,60岁,住院号:33932。该患于半年前出生胃区不适、疼痛,以空腹时为重,同时伴有反酸,无恶心呕吐,饭后有腹胀。曾用胃必治药物治疗二个月无明显好转,近日上述症状加重。既往史:1983年因左脚(足母)趾患恶性黑色素瘤1985年左腹股沟处恶性黑色素瘤传移而在当地医院手术切除。查体:一般状态良好,全身浅表淋巴结无肿大,皮肤、糖膜无异常所见。心肺正常,上腹部有压痛,未触及肿物,肝脾未触及。 X线像见胃窦前壁有3.0×2.5cm大小龛影,周围有环堤,粘膜皱壁破坏中断。诊断为     

恶性黑色素瘤乳腺转移一例

患者女,41岁,因自查发现右乳肿物2个月余就诊于我院.患者于1年前因右足小趾黑痣破溃伴右腹股沟淋巴结肿大入院手术治疗,行右足第4、5趾截趾术、右腹股沟淋巴结清扫术和左腹股沟淋巴结活检术.术后病理报告:右足小趾恶性黑色素瘤,右腹股沟淋巴结1/8(+),左腹股沟淋巴结阴性.术后给予CVD方案[氮烯咪胺(DTIC)+顺铂(DDP)+长春新碱(VCR)]化疗,共6个周期.入院查体结果:右乳肿物无压痛,大小约2.0 cm×1.5 cm,位于外下象限,质硬,表面欠光滑,边界清楚,活动度欠佳;右腋下未触及肿大淋巴结;心、肺、腹及全身检查余无异常.     

MicroRNA与黑色素瘤转移相关性研究进展

黑色素瘤（melanoma）是一种侵袭性强的皮肤肿瘤,近年来随着发病率明显上升,逐渐成为常见恶性肿瘤。其进展迅速,极易转移,临床预后极差,严重危害人类健康。早期黑色素瘤主要通过手术切除治疗,但是对于晚期转移性黑色素瘤,目前治疗手段收效甚微。转移性黑色素瘤对放化疗疗效不明显,     

恶性黑色素瘤鼻咽部转移一例并文献复习

 目的　进一步了解继发性鼻咽部转移瘤的临床生物学特点,避免漏诊误诊。方法　对1例皮肤恶性黑色素瘤鼻咽部转移病例进行分析,并复习相关文献。结果　继发性鼻咽部转移瘤的临床特点和原发性鼻咽部恶性肿瘤极其相似。结论　继发性鼻咽部转移瘤极其罕见,容易误诊为原发性鼻咽部恶性肿瘤,需要通过病史、鼻咽磁共振成像、鼻咽镜、病理活组织检查及免疫组织化学来鉴别与确诊。     

保尔佳对黑色素瘤细胞转移能力及相关细胞行为的影响

考察保尔佳对恶性肿瘤转移的影响及其相关机制。采用细胞微管吸吮实验技术单细胞定量观察不同浓度的保尔佳处理小鼠黑色素瘤细胞不同时间后，瘤细胞对培养牛肺动脉内皮细胞粘附性的改变，同时采用Ｂｏｙｄｅｎ小室法检测经同样处理的瘤细胞的侵袭穿膜能力。采用瘤细胞注射法制作肺转移荷瘤小鼠模型，观察经处理瘤细胞肺转移结节形成数目的改变。保尔佳处理可明显降低瘤细胞与内皮细胞的粘附性，同时抑制其侵袭穿膜能力，这种作用在１０－２～１０μｇ／ｍｌ浓度范围呈剂量依赖性，在１２～４８ｈ范围呈时间依赖性，短时间（３０ｍｉｎ）处理无明显作用。经保尔佳处理的癌细胞，其在肺组织中形成转移结节的数目明显减少。保尔佳通过抑制瘤细胞转移相关性细胞行为—粘附、侵袭，从而起抑制肿瘤转移的作用     

原发性肺恶性黑色素瘤并多脏器转移1例报道

恶性黑色素瘤是起源于成色素细胞的恶性肿瘤,可发生于人体任何部位,皮肤恶性黑色素瘤较多见,而肺原发性恶性黑色素瘤(primary malignantmelanoma of the lung,PMML)较罕见,仅占肺部肿瘤的0.01％[1].国内公开报道符合PMML诊断标准者共33例[2],本院于201 1年收治1例,经皮肺穿刺病理检查确诊为原发性肺恶性黑色素瘤,现探讨该疾病的诊断和治疗,报道如下.     

恶性黑色素瘤伴乳腺转移1例及文献复习

乳腺恶性转移性肿瘤临床少见，占所有恶性乳腺肿瘤的1．79／5～6．6％。常见的乳腺转移性肿瘤有恶性黑色素瘤、肺癌、血液性肿瘤等。本院收治1例并结合国内外1994年～2006年6月报道的临床病理资料完整的39例恶性黑色素瘤伴乳腺转移病例进行分析，报道如下。     

23例恶性黑色素瘤小肠转移临床病例分析

目的：总结恶性黑色素瘤小肠转移的临床病例特点。方法：我院诊治1例,通过中国知网、万方医学网以及 MEDLINE（或 PUBMED）检索到国内外报道的黑色素瘤小肠转移病例共23例,并进行统计分析。收集一般情况、黑色素瘤原发部位、原发肿瘤至小肠转移间隔时间以及临床症状等。结果：23例（男14例,女9例）,平均年龄54.7岁,初次诊断至发现转移平均时间间隔为6.33年（0-20年）。主要临床表现为腹痛16例（69.6%）,黑便/便血7例（30.4%）,肠梗阻或肠套叠14例（60.9%）。肿瘤最大直径平均为5.82cm（2-15cm）。47.8%为小肠多发转移。结论：恶性黑色素瘤小肠转移主要临床表现为腹痛、黑便,易于发生肠梗阻或肠套叠;肿瘤常多发转移,且发现小肠转移距初次诊断时间间隔较长。     

指甲下黑色素瘤术后复发伴肝肺转移1例报告

患者男性，78岁，于1994年无意中发现右拇指指甲中央呈黑褐色，为线条状，无疼痛等不适，未在意。1996年6月发现指甲下有黑色颗粒脱落，指甲中央裂开，甲下有肉芽长出，并逐渐增大。人院体检：右拇指指甲中部呈黑褐色，甲下肉芽组织长出，恶臭，直径约1cm，甲周红肿，甲根部有波动感，腋窝及滑车上淋巴结无肿大。诊断为恶性黑色素瘤。     

Effect of melanoma stem cells on melanoma metastasis

Cancer stem cells (CSCs) are involved in the metastatic process, the resistance of many types of cancer to therapeutic treatments and consequently the onset of recurrences. The CSC concept therefore significantly extends our understanding of melanoma biology. More recently, melanoma stem cells (MSCs) have been described in melanoma as expressing specific biomarkers. These primitive melanoma cells are not only capable of self-renewal and differentiation plasticity, but may also confer virulence via immune evasion and multidrug resistance, and potentially, via vasculogenic mimicry and transition to migratory and metastasizing derivatives. This review will present the specific biomarkers of MSCs, including CD133, ATP binding cassette subfamily B member 5, CD271, CD20 and aldehyde dehydrogenase, which can regulate the transduction of tumor-related signals. These signal molecules can reversely act on tumor cells and regulate tumor angiogenesis, leading to the occurrence of melanoma metastasis. Targeting these specific biomarkers could inhibit the progression of melanoma and may help the development of novel therapeutic strategies for melanoma.     

Timosaponin AIII inhibits melanoma cell migration by suppressing COX‐2 and in vivo tumor metastasis

Melanoma is the leading cause of death from skin disease, due in large part to its propensity to metastasize. We examined the effects of timosaponin AIII, a compound isolated from Anemarrhena asphodeloides Bunge, on melanoma cancer cell migration and the molecular mechanisms underlying these effects using B16‐F10 and WM‐115 melanoma cells lines. Overexpression of COX‐2, its metabolite prostaglandin E₂ (PGE₂), and PGE₂ receptors (EP2 and EP4) promoted cell migration in vitro. Exposure to timosaponin AIII resulted in concentration‐dependent inhibition of cell migration, which was associated with reduced levels of COX‐2, PGE₂, and PGE₂ receptors. Transient transfection of COX‐2 siRNA also inhibited cell migration. Exposure to 12‐O‐tetradecanoylphorbal‐13‐acetate enhanced cell migration, whereas timosaponin AIII inhibited 12‐O‐tetradecanoylphorbal‐13‐acetate‐induced cell migration and reduced basal levels of EP2 and EP4. Moreover, timosaponin AIII inhibited activation of nuclear factor‐kappa B (NF‐κB), an upstream regulator of COX‐2 in B16‐F10 cells. Consistent with our in vitro findings, in vivo studies showed that timosaponin AIII treatment significantly reduced the total number of metastatic nodules in the mouse lung and improved histological alterations in B16‐F10‐injected C57BL/6 mice. In addition, C57BL/6 mice treated with timosaponin AIII showed reduced expression of COX‐2 and NF‐κB in the lung. Together, these results indicate that timosaponin AIII has the capacity to inhibit melanoma cell migration, an essential step in the process of metastasis, by inhibiting expression of COX‐2, NF‐κB, PGE_2, and PGE₂ receptors.     

TNF autovaccination induces self anti-TNF antibodies and inhibits metastasis in a murine melanoma model

TNF is a proinflammatory cytokine involved in the pathogenesis of chronic inflammatory diseases, but also in metastasis in certain types of cancer. In terms of therapy, TNF is targeted by anti-TNF neutralising monoclonal antibodies or soluble TNF receptors. Recently, a novel strategy based on the generation of self anti-TNF antibodies (TNF autovaccination) has been developed. We have previously shown that TNF autovaccination successfully generates high anti-TNF antibody titres, blocks TNF and ameliorates collagen-induced arthritis in DBA/1 mice. In this study, we examined the ability of TNF autovaccination to generate anti-TNF antibody titres and block metastasis in the murine B16F10 melanoma model. We found that immunisation of C57BL/6 mice with TNF autovaccine produces a 100-fold antibody response to TNF compared to immunisation with phosphate-buffered saline vehicle control and significantly reduces both the number (P<0.01) and size of metastases (P<0.01) of B16F10 melanoma cells. This effect is also observed when an anti-TNF neutralising monoclonal antibody is administered, confirming the essential role TNF plays in metastasis in this model. This study suggests that TNF autovaccination is a cheaper and highly efficient alternative that can block TNF and reduce metastasis in vivo and trials with TNF autovaccination are already underway in patients with metastatic cancer.     

Absence of HER2 overexpression in metastatic malignant melanoma

BACKGROUND AND OBJECTIVES: Currently available systemic therapies for malignant melanoma produce low response rates in patients, and more effective treatment modalities are clearly needed. Trastuzumab (Herceptin), the antibody to the HER2 oncoprotein, has had a significant impact on therapy for patients with HER2-overexpressing metastatic breast cancer. This study examined the incidences of HER2 protein overexpression and HER2 gene amplification in metastatic malignant melanoma, which remain unclear in the literature. METHODS: The study evaluated patients with stage III and stage IV malignant melanoma who were treated between 1983 and 1999. Tissue blocks were retrieved and reviewed to confirm the diagnosis. From the 101 cases identified, 49 (31 stage III and 18 stage IV) had sufficient residual tumor sample to enable an assay to be performed. The blocks were tested for HER2 overexpression using the DAKO HercepTest immunohistochemical (IHC) assay. Any sample that tested 1+ or greater for HER2 expression on IHC and a randomly selected subset of HER2-negative samples were tested for the presence of HER2 gene amplification using the Vysis PathVysion fluorescence in situ hybridization (FISH) assay. RESULTS: The median age of the 49 selected patients was 52.2 years, and the male-to-female ratio was 1.23:1 (27 men to 22 women). All of the 49 cases of malignant melanoma were negative for HER2 overexpression by IHC. However, two samples (3%) were found to have a weak level of HER2 expression (1+ level of staining). Subsequent FISH results on the samples that were 1+ on IHC were negative for HER2 gene amplification. FISH results on 21 other randomly selected IHC-negative samples were also negative for HER2 amplification. Flow cytometry failed to show HER2 overexpression in two melanoma cell lines, and treatment of these cells with trastuzumab did not affect their proliferation rate. CONCLUSIONS: We found a low incidence of HER2 expression and no evidence of HER2 protein overexpression or HER2 gene amplification in metastatic malignant melanoma tissues. Therefore, routine testing for HER2 overexpression or HER2 amplification would not be of benefit in this patient population. These results also imply that anti-HER2 therapy with trastuzumab is highly unlikely to provide benefit for patients with metastatic melanoma.     

In vitro and in vivo antitumor effect of 2-methoxyestradiol on human melanoma

2-methoxyestradiol (2ME(2)) is an endogenous metabolite of estradiol with estrogen-receptor-independent antitumor and antiangiogenic activity. We examined the effects of 2ME(2) on the cellular proliferation of 8 human melanoma cell lines. We show that 2ME(2) inhibited cell proliferation by inducing apoptosis and an arrest in the G(2)/M phase, and the mechanism of action involved microtubules, mitochondrial damage and caspase activation. In male SCID mice, 2ME(2) was effective in reducing primary tumor weight and the number of liver metastases after intrasplenic injection of human melanoma cells. In the metastases, we found a significantly higher rate of apoptotic cells after 2ME(2) treatment. These findings on the antitumor effect of 2ME(2) in cell culture as well as in an animal model may have implications for designing alternative treatment options for patients with advanced malignant melanoma.     

The metastatic microenvironment: brain-residing melanoma metastasis and dormant micrometastasis

Brain metastasis occurs frequently in melanoma patients with advanced disease whereby the prognosis is dismal. The underlying mechanisms of melanoma brain metastasis development are not well understood. We generated a reproducible melanoma brain metastasis model, consisting of brain-metastasizing variants and local, subdermal variants that originate from the same melanomas thus sharing a common genetic background. The brain-metastasizing variants were obtained by intracardiac inoculation. Brain metastasis variants when inoculated subdermally yielded spontaneous brain dormant micrometastasis. Cultured cells from the spontaneous brain micrometastasis grew very well in vitro and generated subdermal tumors after an orthotopic inoculation. Expression analysis assays indicated that the brain metastasis and micrometastasis cells expressed higher levels of angiopoietin-like 4, prostaglandin-synthesizing enzyme cyclooxygenase-2, matrix metalloproteinase-1 and preferentially expressed antigen in melanoma and lower levels of claudin-1 and cysteine-rich protein 61 than the corresponding cutaneous variants. The reproducible models of human melanoma metastasizing experimentally and spontaneously to the brain will facilitate the identification of novel biomarkers and targets for therapy and contribute to the deciphering of mechanisms underlying melanoma metastasis.     

Critical role of STAT3 in melanoma metastasis through anoikis resistance

Anoikis is an anchorage-independent cell death. Resistance to anoikis is one of the key features of metastatic cells. Here, we analyzed the role of STAT3 in anoikis resistance in melanoma cells leading to metastasis. When grown under anchorage-independent conditions, significant proportion of cells resisted anoikis and these resistant cells had higher rate of migration and invasion as compared to the cells grown under anchorage-dependent conditions. The anoikis resistant cells also had significantly higher expression and phosphorylation of STAT3 at Y705 than the cells that were attached to the basement membrane. STAT3 inhibitors, AG 490 and piplartine (PL) induced anoikis in a concentration-dependent manner in anoikis resistant cells. Over-expression of STAT3 or treatment with IL-6 not only increased anoikis resistance, but also protected the cancer cells from PL-induced anoikis. On the other hand, silencing STAT3 decreased the potential of cancer cells to resist anoikis and to migrate. STAT3 knock-down cells and PL treated cells did not form tumors as well as failed to metastasize in SCID-NSG mice as compared to untreated anchorage-independent cells, which formed big tumors and extensively metastasized. In summary, our results for the first time establish STAT3 as a critical player that renders anoikis resistance to melanoma cells and enhance their metastatic potential.     

NUAK2: an emerging acral melanoma oncogene

Recent technological advances in cancer genomics make it possible to dissect complicated genomic aberrations of melanomas. In particular, several specific genomic aberrations including 11q13 amplification and KIT aberrations have been identified in acral melanomas. We recently identified NUAK2 at 1q32 as a promising oncogene in acral melanomas and reported its significant roles in tumorigenesis in melanoma cells using both in vitro and in vivo analyses. NUAK2 as a member of the AMPK family has several intriguing aspects both as an oncogene and as a tumor suppressor gene. Here we review genomic aberrations of melanomas focusing on acral melanomas to emphasize the possible roles of NUAK2 in tumorigenesis in general and suggest that NUAK2 has pivotal roles in acral melanomagenesis.