荧光定量PCR法扩增MAGE-1、MAGE-3及AFP基因联合检测肝癌病人外周血微小转移

目的探讨MAGE-1、MAGE-3和AFP mRNA联合检测肝癌病人外周血中微小转移的临床意义。方法应用荧光定量聚合酶链反应（FQ-PCR）检测86例原发性肝癌病人肝癌组织和外周血中MAGE-1、MAGE-3和AFP mRNA的表达。结果86例原发性肝癌病人肝癌组织中MAGE-1、MAGE-3和AFP mRNA的阳性率分别为34．9％（30／86）、60．5％（52／86）和69．8％（60／86）；所有病人肝癌组织至少表达其中一种mRNA。外周血中阳性率分别为14．0％（12／86）、20．1％（18／86）和33．7％（29／86）。52．3％（45／86）的病人外周血至少检测到一种mRNA，较单一AFP mRNA检测阳性率明显提高（P〈0．05）。25例肝炎肝硬化病人及28例健康志愿者中未见阳性。阳性率与肿瘤TNM分期、门脉瘤栓、远处转移有关（P〈0．05）。与肿瘤大小、数目、分化程度、血清AFP水平无关（P〉0．05）。结论MAGE-1、MAGE-3结合AFP mRNA联合检测肝癌血行微小转移较单一AFP mRNA检测灵敏度高。     

复方中药对小鼠B16黑素瘤细胞酪氨酸酶与MITF mRNA表达的调节

目的：探讨复方中药＂白癜冲剂＂治疗白癜风的分子学作用机制。方法：采用定量逆转录-聚合酶链式反应（QRT-PCR）方法,检测＂白癜冲剂＂组（实验组）与无中药组（对照组）小鼠B16黑素瘤细胞株酪氨酸酶与MITFmRNA表达量。结果：实验组黑素瘤细胞酪氨酸酶与MITFmRNA表达量比对照组大（P〈0.05）。结论：复方中药＂白癜冲剂＂能上调黑素细胞酪氨酸酶与MITFmRNA表达水平,可能是其治疗白癜风的作用机制之一。     

肾细胞癌及尿路移行细胞癌组织中黑色素瘤抗原基因mRNA的表达

目的　评价肿瘤特异性黑色素瘤抗原 (MAGE) 1、 3基因作为肾细胞癌及尿路移行细胞癌组织中的免疫学检测和免疫治疗、基因治疗分子标志物的可行性。　方法　肾细胞癌组织标本 18例 ,尿路移行细胞癌组织标本 2 6例 ,相应的癌旁组织 10例 ,采用RT PCR方法对MAGE 1、MAGE 3基因进行测定。　结果　 18例肾癌组织中MAGE 1mRNA阳性表达 10例 (5 6 % ) ,MAGE 3mRNA阳性表达11例 (6 1% ) ,MAGE 1及MAGE 3同时表达 8例 (44 % ) ;2 6例尿路移行细胞癌组织中MAGE 1mRNA阳性表达 16例 (6 2 % ) ,MAGE 3mRNA阳性表达 15例 (5 8% ) ,MAGE 1及MAGE 3同时表达 12例 (46 % )。癌旁组织 10例均不表达MAGE 1及MAGE 3。　结论　MAGE 1、MAGE 3基因有可能作为肾癌及尿路移行细胞癌组织免疫学检测的分子标志物 ,并且具有作为免疫治疗、基因治疗特异性靶位的潜在价值。     

Prognostic significance of isolated HMB45 or Melan A positive cells in Melanoma sentinel lymph nodes

The detection of micrometastases (defined as groups of malignant cells) in the sentinel lymph node (SLN) is an important prognostic tool in melanoma. The use of immunohistochemistry with melanocytic markers such as HMB45 and Melan A increases the detection rate of micrometastases but there are also cases with isolated immunohistochemically positive cells (IPC). To determine the prognostic significance of isolated HMB45 and/or Melan A positive cells in melanoma SLN, we compared the clinical course of 47 patients with IPC to 308 patients with negative SLN and to 122 patients with micrometastases. The mean follow-up was 38.1 months. By Kaplan-Meier analyses, relapse free survival and overall survival of patients with IPC were similar to SLN negative patients, whereas patients with micrometastases had a significantly worse relapse free survival and overall survival. In the 47 patients with IPC, 6 relapses (12.8%) and 3 melanoma-related death (6.4%) occurred, in the SLN negative patients 36 relapses (11.7%) and 17 melanoma-related deaths (5.5%), in the patients with micrometastases 46 relapses (37.7%) and 29 melanoma-related deaths (23.8%). Prognosis of patients with IPC in SLN did not correlate with type of positive staining (HMB45, Melan A, or both), capsular involvement, number of cells, presence of cytologic atypias of IPC, or tumor penetrative depth. In conclusion, with short-term follow-up IPC in melanoma SLN are without prognostic significance.     

Multiple malignant melanomas in association with neurofibromatosis type 1.

We present a case of multiple primary malignant melanomata occurring over a six year period in a 63-year-old Caucasian man with neurofibromatosis type 1. There is doubt regarding a definite association between these two diseases despite a number of case reports and clear, potential pathological mechanisms. This case not only strengthens support for an association but also highlights the great difficulties that arise in the management of cutaneous melanomata in patients with neurofibromatosis.     

HMB-45/melan-A and smooth muscle actin-positive clear-cell epithelioid tumor arising in the ligamentum teres hepatis: additional example of clear cell 'sugar' tumors

HMB-45-positive clear-cell epithelioid tumor arising in the ligamentum teres hepatis of a 13-year-old Japanese girl is described. The well-defined tumor was completely removed and measured 9 x 7 x 6 cm. Cut sections showed a tan-white, homogeneous appearances with no hemorrhage or necrosis. The tumor was composed of nests or sheets of polygonal or oval-shaped cells rich in clear or finely granular cytoplasm. Capillary network was well developed, and sinusoid vessels were often seen with occasional perivascular hyalinization. There was moderate nuclear atypia but mitotic figures were absent. Periodic acid-Schiff stain showed a large amount of glycogen digested by diastase. Immunohistochemical stains for smooth muscle actin, Melan-A, and HMB-45 were positive in most of the tumor cells. Stains for vimentin, muscle actin, and HAM56 were focally positive, whereas stains for desmin, cytokeratin, epithelial membrane antigen, S-100, CD34, CD68, CD99, neurofilament proteins, and estrogen/progesterone receptors were negative. Ultrastructurally, the cytoplasm contained a considerable number of mitochondria, monoparticipate or membrane-bound glycogen, and longitudinally oriented thin filaments with focal condensations and subplasmalemmal densities. The histopathology of the present case, originally interpreted as epithelioid leiomyoma, was consistent with clear cell "sugar" tumors. The present case may indicate ubiquitous distribution of clear cell "sugar tumors" of which histogenesis remains unknown but is presumed to be of perivascular epithelioid cell origin.     

MAGE-1基因在肝细胞肝癌中的表达及临床意义

目的:研究MAGE-1基因在肝细胞肝癌组织中的表达,结合临床资料分析,探讨MAGE-1基因与肝细胞肝癌(HCC)患者临床指标及转移与复发的关系,为MAGE-1基因编码蛋白用于HCC患者免疫治疗提供依据。方法:用RT-PCR的方法对31例HCC患者癌组织及相应癌旁组织MAGE-1基因表达进行检测,随机对6例RT-PCR扩增产物中目的基因片段进行DNA测序以证实其为MAGE-1基因,所有患者均测定并统计AFP、AFU、抗HCV、HBsAAg、AFPmRNA、肿瘤直径等临床指标。结果:31例HCC患者肝癌组织中MAGE-1基因表达的阳性率64.5％(20/31)明显高于癌旁组织2％(1/31),P<0.01。HCC患者肝癌组织中MAGE-1基因表达的阳性率与患者AFP、AFU、抗HCV、乙肝标志物、AFPmRNA、肿瘤直径等临床指标均无关,P>0.05。结论:MAGE-1基因在HCC患者肝癌组织中特异高表达,HCC患者肝癌组织中MAGE-1基因表达的阳性率与HCC患者AFP、AFU、AFPmRNA肿瘤转移、复发均无关。     

Feasibility of integration of modalities in melanomas and sarcomas.

Chemotherapy was administered in the immediate postoperative period to seventy patients (52 with melanomas and 18 with sarcomas) after a total of eighty-seven major operations, with no morbidity or mortality traceable to the chemotherapy. There was no apparent interference with wound healing or what would be considered a normal postoperative course. Fourteen of these patients (5 with melanomas and 9 with sarcomas) received a combination of radiation anc chemotherapy initiated in the postoperative period, and it was tolerated well. This combination appears to be safe, provided the field of radiation is not so large that is may add significantly to the myelosuppressive effect of chemotherapy and the dosage of concomitantly administered radiopotentiating agent(s) is reduced. Sixteen patients had Bacillus Calmette-Gúerin (BCG) immunotherapy in the immediate postoperative period without complications. This policy of a tight interweaving of modalities is safe, has the theoretic advantage of an earlier concerted attack on microscopic residual tumor, and appears particularly promising in sarcomas.     

Anti-alpha 1-antichymotrypsin staining of 194 sarcomas, 38 carcinomas, and 17 malignant melanomas. Its lack of specificity as a tumour marker

A polyclonal commercially available antiserum to alpha 1-antichymotrypsin (alpha 1ACT) was reacted with 194 sarcomas, 38 carcinomas, and 17 malignant melanomas; 61 of 194 sarcomas, 14 of 38 carcinomas, and 10 of 17 malignant melanomas reacted positively. Thirteen categories of sarcomas were examined. All but one group (haemangiopericytomas) showed positively staining tumour cells. Malignant fibrous histiocytomas, angiosarcomas, and Kaposi's sarcomas showed positive staining in over 70% of tumours. All categories of carcinomas examined, with the exception of transitional cell carcinomas, stained positively. The results of this study suggest that the use of alpha 1ACT antiserum is of little value in the differential diagnosis of either sarcomas or carcinomas, nor can it be used as a specific histiocytic marker.     

Induction of MAGE-3 expression in lung and esophageal cancer cells

BACKGROUND: Although MAGE-3 has been detected in approximately 40% of lung and esophageal cancers, expression of this cancer testis antigen appears to be below the threshold for immune recognition in patients with these malignancies. The aim of this study was to determine if the demethylating agent, 5-Aza-2'-deoxycytidine (DAC) and if the histone deacetylase inhibitor Depsipeptide FR901228 (DP) could enhance MAGE-3 expression in lung and esophageal cancer cells. METHODS: Eleven lung and esophageal cancer lines and cultured normal human bronchial epithelial (NHBE) cells were exposed to normal media (NM), DAC, DP, or combination DAC/DP at varying concentrations and exposure durations. MAGE-3 expression was evaluated by quantitative RT-PCR (TaqMan) and immunohistochemistry techniques. Trypan blue exclusion techniques were used to examine the proliferation of cancer cells after drug exposure. RESULTS: Relative to untreated controls, MAGE-3 expression was enhanced 32-fold (range 3.9 to 110) by DAC alone (0.1 micromol/L x 72 h), 2.1-fold (0.4 to 4.2) by DP alone (25 ng/mL x 6h), and 57-fold (4.6 to 209) by sequential DAC/DP exposure. Increased MAGE-3 mRNA copy numbers coincided with enhanced protein levels in these cells. MAGE-3 expression persisted after drug exposure. Flow cytometry confirmed the presence of functional HLA class I expression in these cells. Sequential DAC/DP treatment mediated pronounced growth inhibition in cancer cells but not NHBE. CONCLUSIONS: Sequential DAC/DP treatment may be a novel strategy to simultaneously augment MAGE-3 expression and induce growth arrest in thoracic malignancies.     

应用巢式RT-PCR联合检测原发性肝癌病人外周血AFPmRNA和MAGE-1mRNA的表达

目的 寻找一种敏感的方法以早期发现原发性肝癌的复发与转移。方法 应用逆转录聚合酶链反应分别在术前、术后1周和术后2个月检测37例行根治性切除的原发性肝癌病人外周血中MAGE-1 mRNA和AFPmRNA的表达，并随访所有病人1年。结果 外周血中MAGE-1 mRNA和AFPmRNA的阳性率和肿瘤的病理分期相关；肿瘤病期越晚，外周血中微转移灶的检出率越高。1年的随访中，有9例病人出现复发，MAGE-1和(或)AFPmRNA的阳性率为77．8％(7／9)，其中6例外周血中MAGE-1mRNA和(或)AFPmRNA持续阳性，2例由阴性转为阳性。另有10例病人MAGE-1mRNA和(或)AFPmRNA由术前阳性术后转为阴性，随访1年中无复发。结论 联合检测外周血MAGE-1和AFPmRNA的表达是早期发现肝癌术后复发、评估预后一项敏感可信的指标。     

Clinical Significance of Melanoma Antigen-Encoding Gene-1 (MAGE-1) Expression and its Correlation with Poor Prognosis in Differentiated Advanced Gastric Cancer

Background

Melanoma antigen-encoding gene-1 (MAGE-1), a cancer/testis antigen, has been reported to be expressed in various types of cancer. We investigated the clinicopathological features and prognostic significance of MAGE-1 expression in advanced gastric cancer (AGC).

Methods

Immunohistochemical staining for MAGE-1 was performed on surgical specimens obtained from 135 patients with AGC.

Results

Positive expression of MAGE-1 detected in cytoplasm was observed in 44 of 135 cases (32.6%) in primary tumors and 26 of 96 (27.1%) in lymph node metastases. In noncancerous gastric tissues, apparent MAGE-1 expression was not detected. MAGE-1 in primary tumor was correlated with advanced age (P < 0.001), macroscopic infiltrated type (P = 0.035), and presence of vascular invasion (P = 0.027). The 5-year cancer-specific survival rates of AGC patients with positive MAGE-1 expression were significantly lower than those of patients with negative MAGE-1 (positive: 31.6%, negative: 57.6%, P = 0.038). On multivariate analysis, MAGE-1 expression was not an independent prognostic predictor of AGC (P = 0.064). In differentiated AGC patients, MAGE-1 expression was correlated with advanced age (P = 0.003), macroscopic infiltrated type (P = 0.009), and presence of lymph node metastasis (P = 0.033). The cancer-specific survival rates of differentiated AGC patients with positive MAGE-1 were significantly lower than those of patients with negative MAGE-1 (P = 0.003). Positive MAGE-1 expression was an independent prognostic factor of differentiated AGC patients on multivariate analysis (P = 0.031).

Conclusions

These findings suggest that MAGE-1 protein expression can serve as a predictive marker of poor prognosis in differentiated AGC patients.     

Surgical management of primary cutaneous melanomas of the hands and feet.

OBJECTIVE: The purpose of the study was to investigate the surgical management of cutaneous melanomas of the hands and feet. SUMMARY BACKGROUND DATA: Prior studies suggest that patients with melanomes > 1-mm thick should be treated with excision with a 2-cm margin and undergo elective lymphadenectomy in selected circumstances. These recommendations are based primarily on data from melanomas of the trunk and extremities. Melanomas of the hands and feet are less common and less well studied. They pose a surgical challenge because primary wound closure often is difficult, and the incidence and management of regional node metastases are unclear. METHODS: Charts of patients with melanomas of the hands or feet treated at the Massachusetts General Hospital between 1980 and 1994 were reviewed retrospectively. Local recurrence rates and the incidence of regional node metastases were analyzed as a function of histology, margin of excision, and microscopic thickness of the melanoma. RESULTS: Data from 116 patients (39 men, 77 women) with melanomas of the hands (n = 26) and feet (n = 90) were evaluated. Pathologic diagnoses were: acral lentiginous melanoma (48 patients); subungual melanoma (13 patients), and skin of dorsum of the hand or foot (n = 55). Digital amputation was required in all 13 patients with subungual melanoma to maintain local control; still, nodal metastases developed in 46% of patients within 1 year. Seventy-one percent of patients with acral lentiginous melanoma presented with lesions > or = 1.5 mm, and nodes or systemic disease or both developed in 56% of patients. Acral lentiginous melanoma lesions < 1.5-mm thick were treated principally by excision with a 1-cm margin; a local recurrence or metastases did not develop in any of the patients. None of the patients with melanomas on the dorsum of the hand or foot < 1.5-mm thick had a local recurrence, but regional or systemic disease developed in > 50%. Local control in patients with lesions > 1.5-mm thick frequently required skin grafting or amputation. The majority of patients with melanomas > or = 1.5 mm in thickness undergoing elective lymph node dissection had histologically positive nodes for melanoma. CONCLUSIONS: Melanomas of the hands and feet < 1.5-mm thick have a low incidence of nodal metastases and are treated effectively with wide excision of the primary with a 1-cm margin. Thicker melanomas are associated with a > 50% rate of regional or systemic failure. In the absence of metastatic disease, these individuals should undergo local excision with a 2-cm margin and intraoperative lymphatic mapping followed by lymphadenectomy if the sentinel node is positive.     

Loco-regional renewal of malignant melanomas. I. Local recurrence satellites and in-transit nodes.

183 patients with malignant melanoma treated during the period 1972-1987 were studied for the incidence of loco-regional metastasis. All cases were followed up for 5 years following initial treatment. In the "high risk" groups of patients--the setting being based on well-known prognosticators such as tumour invasion, thickness, clinicopathological type and anatomic site--there were significantly more recurrences (p < 0.01) than in the patients with thin lesions. About 91 per cent of the recurrences--except pure model ones to be discussed in an other paper--were discovered within 30 months following tumour excision. Wide excision in itself had no effect upon frequency and incidence of loco-regional renewal. While local recidivae were sufficiently controlled by simple re-excision, most of the "in-transit" metastases need an additional regional lymph node dissection because of the increased rate of micrometastases in the lymph nodes. Survival and treatment modalities are also discussed.     

人源Dickkopf1原核表达产物对体外培养的黑素细胞的抑制作用

目的:观察人源Dickkopf1原核表达产物(DKK1蛋白)对体外培养的黑素细胞的抑制作用。方法:采用MTT法测定细胞增殖情况;光学显微镜观察细胞形态学变化;氢氧化钠裂解法测定黑素合成;多巴氧化法测定酪氨酸酶活性。结果:DKK1蛋白对黑素细胞的增殖有抑制作用,能使细胞数明显减少(P<0.05);光学显微镜观察到DKK1蛋白作用的黑素细胞胞体较大,形态略呈多角形,树突较粗短;并能使黑素合成显著下降(P<0.05);使酪氨酸酶活性显著减弱(P<0.05)。结论:DKK1蛋白能抑制黑素细胞增殖、黑素合成、酪氨酸酶活性,为DKK1蛋白应用于治疗色素增多性疾病奠定实验基础。