微卫星不稳定在遗传性非息肉病性结肠直肠癌诊治中的作用

遗传性非息肉病性结肠直肠癌（hereditary nonpolyposis colorectal cancer,HNPCC）是最常见的遗传性结肠直肠癌。最新研究表明,微卫星不稳定性（microsatellite instablility,MSI）与HNPCC的发病机制有关,HNPCC病人MSI阳性率高,可作为一种筛查方法,MSI阳性的HNPCC病人,其预后判断也较好;同时,MSI也可指导HNPCC病人的临床治疗。现就MSI在HPNCC中的发病机制、筛选诊断以及预后判断的重要作用作一阐述。     

微卫星不稳定状态对散发性直肠癌预后的影响

目的 研究汉族人散发性直肠癌的微卫星不稳定状态(microsatellite instability,MSI)及其与临床病理学和预后的关系.方法 应用PCR扩增患者肿瘤组织和对应正常组织的DNA,分析微卫星不稳定状态,并采用单因素、多因素分析评估预后.结果 MSI分为高频率微卫星不稳定(microsatellite instability.high,MSI-H)、低频率微卫星不稳定(microsatellite instability-low,MSI-L)、微卫星稳定(microsatellite stability,MSS).93例直肠癌患者中,MSI-H 11例(12%),MSI-L和MSS分别为23例(25%)和59例(63%).与MSI-L和MSS相比,大多数MSI-H为高分化肿瘤、患者年龄小、术前CEA水平高、总的生存率高.结论 在汉族人群散发性直肠癌中,MSI-H发生率高,且MSI是一个相对较好的预后预测指标.     

多重荧光PCR方法对结直肠癌进行微卫星不稳定检测及其临床价值

目的探讨多重荧光PCR方法检测结直肠癌微卫星不稳定（MSI）及其临床意义。方法将2004-2005年间进行手术治疗的110例结直肠癌患者建立队列，以多重荧光PCR方法检测结直肠癌MSI，并对MSI和微卫星稳定（MSS）结直肠癌患者的临床病理特点进行比较。结果多重荧光PCR方法扩增出所有患者的5个微卫星序列。其中MSI．H10例（8．1％），MSI-L13例（11．8％），MSS为87例（79．1％）。共检测BAT．26变异9例（8．2％）、BAT．25变异11例（10．0％）、D2S123变异11例（10．0％）、D5S346变异6例（5．5％）和D17S250变异8例（7．3％）。MSI-L、MSI．H和MSS组结直肠癌患者年龄比较，差异有统计学意义（P〈0．05）；其他临床病理特点差异无统计学意义（P〉0．05）。结论多重荧光PCR方法检测MSI结果稳定，宜于临床应用；MSI和MSS结直肠癌患者临床病理特点比较未见差异。     

胃癌病人微卫星不稳定性发生与DNA倍体的关系

微卫星不稳定性（microsatellite instability．MSI）作为表征基因组不稳定性的一个指标．具有判断预后和指导治疗的重要临床价值。但对胃癌中MSI的研究报道尚不一致，我们通过检测已发现MsI在胃癌病人的胃液中有较高阳性率，且与预后相关。为进一步比较胃癌组织与胃液中MsI的阳性表达率，我们对60例病人的胃癌组织MSI两位点和DNA倍体类型进行了检测。并探讨其与胃癌临床病理参数、DNA倍体类型之间的关系。     

散发性胆管癌患者染色体3p21.3区段微卫星位点不稳定性分析

目的研究散发性胆管癌患者染色体3p21．3区段的微卫星不稳定性（MSI）及杂合性缺失（LOH），探讨染色体3p21．3区段遗传不稳定性与散发性胆管癌发生发展的关系，定位该区段上散发性胆管癌相关肿瘤基因。方法用聚合酶链反应一单链构象多态性分析（PCR—SSCP）方法检测24例散发性胆管癌患者染色体3p21．3区段上D3S1568、D3S1621、D3S1578和D3S1289四个微卫星位点的MSI和LOH发生率，分析其与临床病理因素之间的关系。结果24例散发性胆管癌组织中，4个微卫星位点的MSI和LOH平均发生率分别为7．23％和15．63％。其中D3S1621位点的LOH最高（45．83％，11／24），并与TNM分期、是否伴有局部／淋巴结转移相关（P〈0．05）。结论染色体3p21．3区段133S1621位点高频率杂合性缺失，提示3p21．3区段定位有散发性胆管癌的候选抑癌基因，并在散发性胆管癌的发生发展过程中发挥重要作用。     

微卫星不稳定、血管内皮生长因子表达情况与大肠癌患者病理特征及预后相关性分析

目的探讨分析微卫星不稳定(MSI)、血管内皮生长因子(VEGF)表达情况与大肠癌患者病理特征及预后相关性。方法回顾性分析本院107例行大肠癌根治术治疗的大肠癌患者临床病理资料,采用免疫组织化学方法检测107例患者手术标本肿瘤组织内MSI及VEGF表达情况,并比较MSI、VEGF表达与患者病理特征、预后相关性。结果 MSI与患者淋巴结转移与浸润深度相关(P<0.05);VEGF表达与患者血行转移、淋巴结转移、TNM分期及浸润深度相关(P<0.05);高微卫星不稳定性(MSI-H)大肠癌VEGF的表达显著减少。结论MSI与大肠癌患者淋巴结转移与浸润深度相关,而VEGF表达与大肠癌患者血行转移、淋巴结转移、TNM分期与浸润深度相关,MSI-H患者其生存及预后情况显著优于MSS及MSI-L患者;VEGF阴性患者其生存情况显著优于VEGF阳性患者。MSI-H大肠癌与微卫星稳定性(MSS)大肠癌可能存在两种不同的新生血管形成的途径,MSI-H肿瘤较低的VEGF表达或许可解释为何MSI-H大肠癌有较低的侵袭性。     

微卫星不稳定对散发性结直肠癌患者预后的影响

目的 探讨微卫星不稳定(MSI)对散发性结直肠癌预后的影响.方法 收集2004年8月至2006年9月南京中医药大学第三附属医院手术治疗并具有完整随访资料的134例结直肠癌病例,根据MSI检测结果 将其分成MSI组和微卫星稳定(MSS)组.采用单因素和多因素预后分析来评估MSI的预后价值.结果 134例患者中MSI组26例,MSS组108例.两组患者术后复发率分别为7.7%(2/26)和35.2%(38/108),差异有统计学意义(P=0.006);两组患者5年生存率分别为92.3%和63.5%,差异亦有统计学意义(P=0.016).经多因素分析,MSI为结直肠癌患者的独立预后因素(P=0.029).结论 微卫星不稳定是影响散发性结直肠癌患者预后的重要因素.

Abstract：

Objective To investigate the role of microsatellite instability (MSI) in Chinese sporadic coloretal cancer. Methods A total of 146 patients with colorectal cancer were treated surgically from August 2004 to September 2006 in the Third Affiliated Hospital of Nanjing University of Traditional Chinese Medicine. Data were collected prospectively. Univariate and multivariable analyses were performed for parameters such as age, gender, tumor location, differentiation, MSI, tumor type, lymph node metastasis,TNM stage, and survival. Results Follow-up was available in 134 patients including telephone call and office visit. MSI(P=0.029), tumor type (P=0.000), TNM stage (P=0.000) were independently associated with survival on Cox regression model. There were 26 patients with MSI, and the 1-, 3-, and 5-year survival rates were 100% , 92.3% , and 92.3% , respectively. The remaining 108 patients had microsatellite stable tumor, and the 1-, 3-, and 5year survival rates were 96.3% , 72.2% , and 63.5% , respectively. The difference was statistically significant(P=0.016). Conclusion Microsatellite instability is an important factor associated with patient survival in Chinese sporadic colorectal cancer.     

转化生长因子βⅡ受体、胰岛素生长因子Ⅱ受体、凋亡诱导蛋白BAX和转录因子E2F4基因移码突变与胃癌微卫星不稳定性的关系

目的探讨胃癌组织中与细胞增殖、凋亡和转录有关的基因移码突变与微卫星不稳定性（MSI）的关系。方法采用酚．氯仿一异戊醇法从胃癌及切缘正常石蜡组织中提取DNA。组织切片中肿瘤成分不足50％时采用显微切割方法。以聚合酶链反应一单链构象多态性（PCR-SSCP）、直接DNA测序法检测MSI及转化生长因子BII受体（TGFβRⅡ）、胰岛素生长因子Ⅱ受体（IGFⅡR）、凋亡诱导蛋白BAX和转录因子E2F4基因的移码突变。按照MSI的发生频率将胃癌患者分为3组：出现≥2个位点的MSI为高MSI,出现1个位点的MSI为低MSI,无MSI发生为微卫星稳定。结果61例胃癌患者中,12例（19．7％）为高MSI,11例（18．0％）为低MSI,38例（62．3％）为微卫星稳定。TGFβRⅡ、IGFⅡR、BAX和E2F4移码突变检出率分别为12例（19．7％）、3例（4．9％）、4例（6．6％）和10例（16．4％）。12例高MSI胃癌中,有10例TGFβRⅡ基因突变,3例IGFⅡR基因突变,4例BAX基因突变,10例E2F4基因突变;其移码突变率与高MSI发生密切相关。微卫星稳定组的肿瘤中未发现这些基因的突变。结论TGFβRⅡ、IGFⅡR、BAX和E2F4基因编码区重复序列是MSI发生的靶点,这些基因的移码突变在MSI胃癌的发生和发展中起了重要作用。     

胃癌微卫星不稳定性及其与临床病理生物学的关系

目的探讨微卫星不稳定性 (MSI)发生与胃癌临床病理生物学行为的关系及其临床意义.方法采用酚氯仿异戊醇法从胃癌组织切片中提取 DNA.组织切片中肿瘤成分不足 50%时采用显微微切割方法.以 PCR为基础的单链构像多态性方法分析胃癌组织分别在 BAT26、 BAT40、 D2S123、 D5S346和 D17S250 5个位点的微卫星不稳定性.结果按照微卫星不稳定性的发生频率将胃癌患者分为 3组 出现 2个或 2个以上微卫星位点不稳定性为高 MSI组( MSI-H);出现 1个位点不稳定性为低 MSI组( MSI-L);无 MSI发生为微卫星稳定组( MSS). 61例胃癌患者中, 12例( 19.7%)为 MSI-H, 11例( 18.0%)为 MSI-L, 38例( 62.3%)表现为 MSS. MSI-L组与 MSS组患者在胃癌各临床病理生物学参数间差异无显著性意义( P >0.05),但这两组与 MSI-H组比较,在肿瘤的发生位置、 Lauré n分型、淋巴结是否转移和肿瘤分期方面差异有显著性意义( P< 0.05). MSI-H型胃癌多发生于胃窦部,多见于淋巴结转移阴性、肠型和较早期胃癌,其发生与患者年龄、性别和肿瘤大小无明显的相关性.结论 MSI-H型胃癌与 MSI-L或 MSS型胃癌具有不同的临床病理学行为, MSI-H型胃癌与肿瘤位置、淋巴结是否转移、 Lauré n分型及肿瘤分期密切相关.微卫星不稳定性可能是胃癌发生的另一分子机制.     

微卫星不稳定结直肠癌hMLH1和hMSH2及hMSH6种系突变与hMLH1启动子甲基化检测

目的探究微卫星不稳定（MSI）结直肠癌患者的hMLH1、hMSH2和hMSH6种系突变特征和hMLH1启动子甲基化状态。方法对前瞻性收集的34例MSI结直肠癌患者检测其hMLH1、hMSH2和hMSH6种系突变，并研究其肿瘤的hMLH1启动子甲基化状态。结果34例MSI结直肠癌中。共检测到MLH1基因启动子的甲基化19例（55．9％）。19例MSI—H结直肠癌中检测到MLH1基因的甲基化14例（73．7％）；15例MSI—L结直肠癌检测到MLH1基因的甲基化5例（33．3％）；两组差异有统计学意义（P〈0．05）。全组共发现8个hMSH2和hMSH6基因的突变，其中hMSH6基因突变3个，hMSH2基因突变5个。结论中国人MSI结直肠癌错配修复基因突变（未测到MLH1基因突变）和MLH1基因启动子的甲基化检出率可能有别于国外MSI结直肠癌。     

肿瘤浸润淋巴细胞在结直肠癌中的研究进展

随着肿瘤免疫学的研究进展,人们发现人体免疫系统抗肿瘤的作用更能决定患者的预后。结直肠癌问质中肿瘤浸润淋巴细胞（TIL）与结直肠癌的关系成为目前研究的热点之一。本文总结了近年来关于不同亚型肿瘤浸润T淋巴细胞与结直肠癌的临床病理因素、预后及微卫星不稳定性（MSI）的关系。     

乳腺癌与微卫星不稳定性的相关研究

目的 探索乳腺癌微卫星不稳定性(MSI)与临床特征的相关性.方法 收集60例乳腺癌组织标本,选择5个微卫星多态性标记(位点),DNA的提取,PCR扩增,电泳,EB显色后进行MSI分析.结果 乳腺癌MSI发生率为33.3%,正常乳腺组织MSI发生率为0,乳腺癌MSI与分化程度有关.结论 MSI是乳腺癌发生的早期分子生物学...     

Clinical significance of mutator phenotype and chromosome 17p and 18q allelic loss in gastric cancer

BACKGROUND: Tumour stage is the only reliable prognostic factor for gastric cancer. The molecular anomalies involved in this process have the potential to serve as additional prognostic markers. METHODS: Forty-four gastric cancers, treated by surgery alone, have been analysed for chromosome 17p and 18q allelic loss and for the presence of microsatellite instability (MSI), using microsatellite markers and DNA from paraffin-embedded tumours. RESULTS: Eight cancers showed a MSI-positive (MSI+) phenotype. Among the 36 MSI-negative cancers, chromosome 17p and 18q allelic losses were found in 22 of 34 and 19 of 33 informative cases respectively. Multivariate survival analysis indicated MSI status to be an independent prognostic factor along with the tumour stage. MSI+ cancers were associated with longer patient survival, whereas MSI-negative cancers had a significantly poorer prognosis (P = 0.007), with a median actuarial survival of 24 months. CONCLUSION: MSI status is an independent prognostic factor among gastric cancers at the same stage. Chromosome 17p and 18q status added no additional prognostic information to that of tumour stage. The combined use of tumour stage and MSI status may help in deciding whether patients with advanced gastric cancer require additional therapy other than surgery alone.     

膀胱癌染色体微卫星不稳定性及杂合性丢失

目的　探讨染色体微卫星不稳定性（ ＭＳＩ） 及杂合性丢失（ＬＯＨ） 在膀胱癌早期诊断中的意义。　方法　运用位于４ 、５ 、９ 、２１ 号染色体上的四对微卫星标志,结合ＰＣＲ 银染技术,分别检测２２例膀胱癌患者肿瘤组织及尿脱落细胞染色体ＭＳＩ及ＬＯＨ。　结果　肿瘤及尿液标本ＭＳＩ的发生率分别为２３ ％ 、１９ ％ ,ＬＯＨ 发生率分别为４１ ％ 、１８ ％ ,至少有一个位点出现ＭＳＩ或ＬＯＨ 分别为５９ ％ 及３２ ％ 。不同年龄、性别、临床分期、肿瘤组织分级的患者ＭＳＩ与ＬＯＨ 的发生率差异均无显著性（ Ｐ＞０ ．０５） 。　结论　检查尿液中肿瘤脱落细胞的ＭＳＩ及ＬＯＨ 可能成为膀胱癌筛选及监测复发的辅助方法。     

Chemosensitivity and Survival in Gastric Cancer Patients with Microsatellite Instability

Introduction  Conflicting data exist regarding the relevance of high-frequency microsatellite instability (MSI-H) for predicting the prognosis and benefits of 5-fluorouracil (5-FU)-based chemotherapy. This study investigated the usefulness of MSI as either a prognostic indicator or predictor of distinct clinical attributes regarding the use of adjuvant chemotherapy with 5-FU and its analogues in gastric cancer. Materials and methods  Data and tumor specimens were collected from 240 gastric cancer patients from 1993 to 2002. Five microsatellite loci were analyzed using a high-intensity microsatellite analysis reported previously. A Cox proportional hazard model was used to compare the clinical data and survival as well as any associations between MSI and 5-FU treatment status of patients with MSI or microsatellite stability (MSS) gastric cancers. A 3-(4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2H-tetrazolium bromide (MTT) assay was conducted in 168 cases to investigate chemosensitivity to 5-FU. Results  This analysis identified 22 MSI-H (9.4%), 25 MSI-L (10.7%), and 193 MSS (79.9%) tumors. Gastric cancer with MSI-H tended to have increased likelihood to show higher age, antral location of the tumor, and lymph vessel involvement (P < 0.05). Univariate analyses failed to show any difference between the MSI-H and MSS/MSI-L groups with respect to overall survival. Furthermore, survival after the administration of 5-FU did not correlate with MSI status, and MSI was not associated with 5-FU sensitivity by MTT assay. Conclusion  The results of this study indicate that MSI status has no clear influence on overall survival or response to 5-FU in gastric cancer.     

Notch信号转导途径配体基因JAG1和DLL1在结直肠癌中的表达及临床意义

目的探讨Notch信号传导途径配体基因JAG1和DLL1在结直肠癌的表达及其与临床病理特征和肿瘤预后之间的关系。方法回顾性分析2004年8月至2006年9月在南京中医药大学第三附属医院全国肛肠医疗中心行手术治疗的146例结直肠癌患者的临床及随访资料．建立患者肿瘤标本组织芯片并对JAG1和DLL1基因表达进行免疫组织化学检测。结果146例患者JAG1表达与其肿瘤分化程度有关．DLL1表达在不同肿瘤部位的表达差异有统计学意义（均P〈0．05）;两种基因表达与微卫星不稳定状态之间差异无统计学意义（P〉0．05）。134例（91．8％）患者获随访,随访时间（42．3±13．3）个月。无瘤生存86例（64．1％）,1、3和5年总生存率分别为93％、74％和67％。多因素分析显示,患者预后与TNM分期、病理学类型、微卫星状态、JAG1基因表达有关（P〈0．05）。JAG1基因高表达患者预后优于阴性表达和弱阳性表达的患者（P〈0．05）。结论Notch信号转导途径配体基因JAG1和DLL1表达分别与肿瘤分化程度及肿瘤部位有关．JAG1基因与预后有关。     

Microsatellite instability in gastric cancer is associated with better prognosis in only stage II cancers

BACKGROUND: The assessment of microsatellite instability (MSI) is not included yet in the routine evaluation of patients with gastric cancer, as controversial data exist regarding its prognostic value. METHODS: We determined the clinical significance of MSI in 510 sporadic gastric cancers, using the mononucleotide markers BAT25 and BAT26. The results were compared with the immunohistochemical expression of the mismatch repair proteins Mlh1 and Msh2. RESULTS: MSI was present in 83 (16%) cancers and correlated with better survival (P < .001). Multivariate analysis showed that the MSI phenotype was an independent factor (P = .005) and added prognostic information to TNM stage, location, and age. The relative risk of death for MSI cancer patients was 0.6 (95% confidence interval [CI], 0.4-0.8). Moreover, when grouped according to stage, only stage II cancers showed a significant effect of MSI status on survival (P = .011; hazard ratio = 0.3; 95% CI, 0.1-0.8). MSI also correlated with older age (P = .002), female gender (P < .001), intestinal histotype (P = .011), lower T stage (P = .018), and less lymph node involvement (P < .001). Finally, comparison of the results of immunohistochemical expression of the mismatch repair proteins Mlh1 and Msh2 with microsatellite analysis showed concordant results in 95% of neoplasms, with a sensitivity of 82% and specificity of 98%. CONCLUSIONS: Microsatellite analysis of gastric cancer has clinical utility in determination of prognosis, but should be determined in only stage II neoplasms in a routine clinical setting. Immunohistochemistry may be considered sufficient, although microsatellite analysis is preferable.     

Microsatellite alterations in various sarcomas in Japanese patients

To determine the usefulness of microsatellite analysis in the differential diagnosis of various sarcomas, we investigated microsatellite alterations at 12 microsatellite loci by polymerase chain reaction and electrophoresis in 39 Japanese patients with sarcomas. The sarcomas were: osteosarcoma, Ewing's sarcoma, chondrosarcoma, liposarcoma, leiomyosarcoma, epithelioid leiomyosarcoma, rhabdomyosarcoma, synovial sarcoma, and malignant fibrous histiocytoma. We also examined ten leiomyomas to contrast with leiomyosarcoma. No microsatellite instability (MSI) or loss of heterozygosity (LOH) were found in Ewing's sarcoma, chondrosarcoma, epithelioid leiomyosarcoma, malignant fibrous histiocytoma, and leiomyoma. Only three patients, one each with liposarcoma, leiomyosarcoma, and synovial sarcoma, manifested MSI, whereas, osteosarcoma, liposarcoma, leiomyosarcoma, rhabdomyosarcoma, and synovial sarcoma manifested LOH, with an incidence of 43%, 14%, 86%, 20%, and 75%, respectively. Interestingly, three patients showed unusual patterns of LOH, probably due to intratumoral heterogeneity. Kaplan-Meier analysis revealed that LOH on 11p was predictive of poor prognosis in osteosarcoma. The low incidence of MSI indicates that MSI is not necessary for neoplastic transformation in sarcomas. However, the very high incidence of LOH in leiomyosarcoma indicates that microsatellite analysis may serve for the differential diagnosis of leiomyosarcoma versus leiomyoma. Microsatellite analysis may also predict prognosis in osteosarcoma. Received for publication on Aug. 14, 1998; accepted on Dec. 21, 1998