[雙語翻譯]--醫(yī)學(xué)外文翻譯--食管癌和賁門癌患者中mage， bage和gage基因的表達(dá)（原文）

1、MAGE, BAGE, and GAGE Gene Expression in Patients with Esophageal Squamous Cell Carcinoma and Adenocarcinoma of the Gastric CardiaAnnalisa Zambon, Ph.D.1Susanna Mandruzzato, Ph.D.1Anna Parenti, M.D.2Beatrice Macino, Ph.D.

2、1Piero Dalerba, M.D.1Alberto Ruol, M.D.3Stefano Merigliano, M.D.3Giovanni Zaninotto, M.D.3Paola Zanovello, Ph.D.11 Department of Oncology and Surgical Sciences, Oncology Section, University of Padova, Padova, Italy.2 Dep

3、artment of Oncology and Surgical Sciences, Pathology Section, University of Padova, Padova, Italy.3 Department of Medical and Surgical Sciences, Clinica Chirurgica 4, University of Padova, Padova, Italy.Presented at the

4、33rd Congress of the European Society for Surgical Research (ESSR), Padova, It- aly, April 22–25, 1998 and at the International Society for the Diseases of the Esophagus (ISDE) Seventh World Congress, Montreal, Quebec, C

5、an- ada, September 1–4 1998.Supported by the Associazione Italiana per la Ricerca sul Cancro (AIRC, Milan, Italy) and the Regione Veneto (grant 657/02/96, Ricerca Sani- taria Finalizzata). S.M. was supported by a post- d

6、octoral fellowship from ICGEB, Trieste, Italy; and B.M. was supported by a personal grant from AIRC.The authors are grateful to Dr. G. L. De Salvo and M. Epifani for statistical analysis and to P. Segato for help in arti

7、cle preparation.Address for reprints: Alberto Ruol, M.D., Clinica Chirurgica 4, University of Padova, Via Giustiniani, 2, 35128 Padova, Italy; Fax: ?39.049.8213152; E-mail: aruol@ux1.unipd.it.Received June 28, 2000; revi

8、sion received Janu- ary 10, 2001; accepted January 19, 2001.BACKGROUND. The MAGE, BAGE, and GAGE gene families code for distinct, tumorspecific antigens that are recognized by cytotoxic T lymphocytes in the context ofHLA

9、 molecules. The purpose of this study was to analyze MAGE, BAGE, and GAGEgene expression in the two major histologic types of esophageal carcinoma, squa-mous carcinoma (ESCc) and adenocarcinoma (CAc), and to correlate th

10、eir expres-sion patterns with the principal prognostic parameters and long term survival.METHODS. Gene expression was analyzed in surgical samples from 24 patients withESCc and 24 patients with CAc by reverse transcripta

11、se-polymerase chain reactionamplification (RT-PCR). None of the patients had received preoperative chemo-therapy or radiotherapy, and all were followed until death or for a minimum of 4years.RESULTS. Sixteen ESCc samples

12、 (67%) and 9 CAc samples (37.5%) expressed at leastone of the genes under study. The expression of each MAGE gene in the twohistologic types was not significantly different, with the exception of MAGE-4,which was express

13、ed more in ESCc samples than in CAc samples. BAGE and GAGEexpression was rather low and, in every case, was associated with the expression ofat least one MAGE gene.CONCLUSIONS. In the group as a whole, and in both ESCc a

14、nd CAc subgroups, nosignificant correlation emerged between the expression of any gene and prognosticparameters, such as pathologic tumor, lymph node, or disease stage. Nevertheless,BAGE or GAGE expression was related si

15、gnificantly to a poor prognosis, whereasthe expression of MAGE genes (in the absence of BAGE and GAGE expression) wasrelated significantly to a good prognosis. Cancer 2001;91:1882–8.© 2001 American Cancer Society.KE

16、YWORDS: esophageal carcinoma, tumor antigens, MAGE, BAGE, GAGE.I n recent years, numerous human tumor antigens that are recog- nized by autologous cytotoxic T lymphocytes (CTLs) have been identified.1 An important catego

17、ry of these so called T-cell-defined tumor antigens consists of normal gene products that are not ex- pressed in most body tissues, with the exception of male germline cells and placenta, and are activated in a number of

18、 different tumors: Antigenic peptides encoded by the MAGE, BAGE, and GAGE gene families are prototypes of this category of shared tumor antigens.2–5Although they initially were described in melanoma, these genes have bee

19、n found to be expressed in a substantial number of solid tumors in various organs, such as lung, breast, bladder, head and neck, esoph- agus, and stomach, as well as in several tumor cell lines2; because of their strict

20、tumor specificity, they are of particular interest for cancer immunotherapy.1882© 2001 American Cancer Societydeoxynucleotides (dNTPs), 1 ?L of a 500-?g/mL solu- tion of oligo(dT)12–18 primers, 20 units of RNaseOUT

21、(GIBCO BRL), 2 ?L of 0.1 M 1,4-dithiothreitol, and 200 units of Moloney murine leukemia virus reverse tran- scriptase (GIBCO BRL); the reaction was incubated at 42 °C for 60 minutes and then diluted to 40 ?L with wa

22、ter. Two microliters of the cDNA mixture were used for each polymerase chain reaction (PCR) amplifica- tion in a 50-?L reaction volume containing 1 ?L of each primer (40 ?M), 1 ?L each of 2.5 mM dNTP, 1.5 mM MgCl2, and 2

23、 units Taq DNA polymerase (Pro- mega, Madison, WI) in Buffer A, which was supplied by the manufacturer. The primers used in this study to ensure specificity for each gene were described previ- ously.3,4,9 Thirty-two ampl

24、ification cycles were run: 1 minute at 94 °C and 3 minutes at 72 °C for MAGE-1 and MAGE-3; 1 minute at 94 °C, 2 minutes at 68 °C, and 2 minutes at 72 °C for MAGE-2 and MAGE-4; 1 minute at 94 

25、6;C, 2 minutes at 70 °C, and 2 minutes at 72 °C for MAGE-6; 1 minute at 94 °C, 2 minutes at 62 °C, and 2 minutes at 73 °C for BAGE; and 1 minute at 94 °C, 2 minutes at 55 °C, and 2 minu

26、tes at 72 °C for all of the GAGE genes. Cycling was concluded with a final extension step of 15 minutes at 72 °C. To verify RNA integrity in each sample, 23 cycles (for 1 minute at 94 °C, 2 minutes at 68 &

27、#176;C, and 2 minutes at 72 °C) were run with primers specific for ?-actin. After amplifica- tion, PCR products were analyzed by agarose gel elec- trophoresis.Statistical Analysis Statistical analyses were performed

28、 using the SAS sta- tistical package (SAS, Inc., Cary, NC). Differences be- tween groups were assessed by chi-square analysis, Fisher exact test, or Student t test, as indicated. All P values ? 0.05 were considered signi

29、ficant. Survival was measured from the date of surgery to death or last date of follow-up. Survival rates and standard errors were calculated by the Kaplan–Meier method, includ- ing deaths from all causes, except the two

30、 hospital deaths that were related to postoperative complica- tions. The statistical significance of differences in sur- vival was analyzed by the log-rank test, with P ? 0.05 considered significant. The prognostic impor

31、tance of clinical variables was evaluated by a Cox proportional hazard regression using multivariate analysis.RESULTS MAGE-1, MAGE-2, MAGE-3, MAGE-4, MAGE-6, BAGE, and GAGE gene expression was evaluated in 48 surgi- cal

32、specimens, of which 24 specimens were ESCc, and 24 specimens were CAc (Table 1), and in 5 samples of normal esophageal tissue adjacent to the tumor. Sixty-seven percent of the ESCc tumors and 37.5% of the CAc tumors expr

33、essed at least one of these genes. Figure 1 shows the different patterns of MAGE-BAGE- GAGE gene expression in eight representative CAc samples together with appropriate controls. Table 2 shows that the pattern of MAGE g

34、ene expression was very similar in both ESCc and CAc samples, except for MAGE-4, which was expressed significantly more in the ESCc samples (58% vs. 25%, respectively; P ? 0.019). Two ESCc samples (8%), but none of the C

35、Ac samples, expressed BAGE (P ? 0.149). GAGE gene ex- pression was observed in four ESCc samples (17%) and in three CAc samples (13%; P ? 0.683). None of the genes under study was expressed in normal esoph- ageal tissue

36、samples (data not shown). We investigated the association between MAGE, BAGE, and GAGE gene expression, and the clinico- pathologic parameters listed in Table 1. In the group of patients as a whole and in both ESCc and C

37、Ac subgroups, no significant correlation emerged be- tween the expression of any of these genes, and pa- tient gender and age, histologic tumor type and grad- ing (pT, pN, and pStage), and type of resection, with the sin

38、gle exception of the GAGE gene, which was expressed significantly more in tumor samples ob- tained from patients who underwent R1–R2 resection (P ? 0.027; data not shown). MAGE, BAGE, and GAGE gene expression appeared to

39、 be clustered in a subset of the tumors examined, because most lesions either did not express any gene (48%; 23 of 48 patients) or simultaneously coexpressed three or more genes (35%; 17 of 48 patients) (Table 3). BAGE a

40、nd GAGE genes always were coexpressed with one or more members of the MAGE family. The overall 1-year, 3-year, and 5-year actuarial survival rates of patients surviving esophagectomy (n ? 46 patients) were 85%, 41%, and

41、24%, respectively. The 1-year, 3-year, and 5-year actuarial survival rates after R0 resection (n ? 37 patients) were 89%, 51%, and 30%, respectively. Table 4 summarizes the univar- iate analysis of the prognostic value o

42、f several clinico- pathologic parameters. In the first set of survival analyses, the samples were divided into two cohorts: those expressing one or more of the genes studied and those showing no gene expression. In the e

43、ntire group of patients and in both ESCc and CAc subgroups, which were evaluated sep- arately, no significant association was found between the above two cohorts and survival (data not shown). A similar analysis was carr

44、ied out for each of the MAGE, BAGE, and GAGE genes separately: Only BAGE or GAGE expression was related significantly to a poor prognosis (Table 4). Finally, we grouped the patients according to the1884 CANCER May 15, 20