良、恶性卵巢肿瘤C－Ha－rasl位点的等位基因型与杂合性缺失

为了探讨Ｃ－Ｈａ－ｒａｓｌ位点的等位基因型与杂合性缺失在卵巢癌发生中的意义，应用Ｓｏｕｔｈｅｒｎ印迹技术，在１７例良、恶性卵巢肿瘤患者中研究Ｃ－Ｈａ－ｒａｓｌ位点的多态性和等位片段缺失。结果表明，在ＢａｍＨＩ酶切、Ｃ－Ｈａ－ｒａｓｌ探针杂交的放射自显影图上显示７．８ｋｂ（Ｌ）和６．８ｋｂ（Ｓ）两个等位片段，基因型ＳＳ的个体在恶性卵巢肿瘤组的比例（７／１１）显著高于良性肿瘤组（２／６）（Ｐ＜０．０１）；在４例正常组织的ＬＳ杂合子患者中，１例患者的卵巢癌细胞出现等位片段杂合性丢失（ｌｏｓｓｏｆｈｅｔｅｒｏｚｙｇｏｓｉｔｙ，ＬＯＨ）和重排，并且有高度恶性的表型。本结果提示：等位片段Ｓ的钝合子和ＬＯＨ后的半合子（ｈｅｍｉｚｙｙｏｔｅ）较等位片段Ｌ纯合子和ＬＳ杂合子有更大的恶性卵巢肿瘤的易感性。     

Molecular pathogenesis of adenoma and differentiated adenocarcinoma of the stomach

Advances In molecular biology have revealed a consistent set of genetic aiterations that may correspond to multistep tumor development. The pathogenesis of adenoma and differentiated adenocarcinoma of the stomach are reviewed from a genetic perspective with reference to the colorectal adenoma-carcinoma sequence. The sequential accumulation of genetic alterations characteristic of the colorectal adenoma-carclnoma sequence does not occur between sdenoma and differentiated adenocarclnoma of the stomach, although adenomatous polyposis coli (Am) mutation in adenoma, and p53 mutation and loss of heterozygosity (LOH) of DCC (deleted In colorectal cancer) gene in carcinoma are prevalent genetlc alterations. Allelotype, LOH and micro-satelllte anatyses have revealed several chromosomal regions of ddetlon, as well as genetic Instability, that accumulate during the development and progression of differentiated adenocarcinomas. However, these alterations are rarely found In adenomas of the stomach. These findings suggest that the adenoma-carcinoma sequence Is relatively rare in gastric carcinogenesis, and that most differentiated adenocarcinomas of the stomach develop through a de novo pathway.     

COLLISION TUMOR OF THE STOMACH WITH CARCINO-SARCOMA AND TUBULO-PAPILLARY ADENOCARCINOMA

The present case is an autopsy case of a 66-year-old male who developed a collision tumor in the stomach which was originated from carcinosarcoma of Borrmann type II in the gastric antrum and from papillary adenocarcinoma of Borrmann type III in the corpus. The carcinosarcoma consisted of tubular carcinoma and sarcomatous transformation of stroma from the carcinoma.     

CLEAR CELL ADENOCARCINOMA OF THE FEMALE URETHRA A Case Report

A case of urethral clear cell adenocarcinoma (mesonephric carcinoma) in a 62-year-old woman is reported. The patient consulted our hospital because of acute urinary retention. Fine needle aspiration cytology showed a few atypical cells; loose cluster cells with finely vacuolated cytoplasm, large nuclei and prominent nucleoli, and small cluster cells with finely vacuolated, delicate cytoplasm, small pale nuclei and small nucleoli. Cytologic diagnosis was adenocarcinoma and no diagnosis of clear cell adenocarcinoma was obtained. The neoplasia was localized at the posterior wall of the urethra and deep muscular layer of the vaginal wall. Histologically, the tumor revealed mainly small, elongated glands consisting of single-layered cuboidal or columnar cells with scanty cytoplasm and a focally hobnail appearance. In addition, a small cystic lesion resembling nephrogenic adenoma was observed. This finding raises the possibitily that clear cell adenocarcinoma may be a malignant counterpart of nephrogenic adenoma. ACTA PATHOL JPN 38: 217–223, 1988.     

人胶质母细胞瘤等位基因谱分析的研究

目的 探讨胶质母细胞瘤（glioblastoma,GBM)发病的分子遗传学机理，确定GBM的发生发展主要和哪些染色体或染色体区域有关，哪些染色体区域上可能存在与GBM相关的肿瘤抑制基因（tumor suppressor gene,TSG)。方法 应用聚合酶链反应技术，采用荧光标记的引物和377型DNA序列自动分析仪，对21例GBM的所有22对常染色体上共计382个微卫星位点进行了杂合性丢失（loss of heterozygosity,LOH)分析，相邻2个微卫星位点之间的平均遗传学距离为10cM。结果 在所有被检测的染色体臂上都观察到LOH，其中以染色体10q、10p、9p、17p和13q的LOH率最高（＞50％），这些染色体臂上已知的肿瘤抑制基因PTEN、DMBT1、p16、p53和Rb所在区域LOH率都较高；14q、3q、22q、11p、9q、19q上也存在较高的LOH率（＞40．5％）；首次发现多个共同微小丢失区域：9p22-23、10p12.2-14、10q21.3、13q12.1-14.1、13q14.3-31、17p11.2-12、17p13、3q24-27、11p12-13、14q31-32.3、14q21-24.1、22q13.2-13.3、4q35、4q31.1-31.2、6qtel、6q16.3。结论 GBM存在复杂的遗传学异常，涉及多条染色体臂。以10q、10p、9p、17p和13q的异常与GBM发生发展的关系最为密切。除了已知的肿瘤抑制基因PTEN、DMBT1、p16、p15、p53和Rb外，首次所发现的多个微小共同丢失区域上可能存在GBM相关的多个未知TSG。     

复发前后多形性胶质母细胞瘤的等位基因谱分析

目的：研究多形性胶质母细胞瘤（glioblastoma multiforme,GBM)复发前后的分子遗传学变化，了解基因组范围内哪些染色体区域可能与GBM复发有关。方法：应用聚合酶链反应技术为基础的杂合性丢失（loss of heterozygosity,LOH)分析法，采用荧光标记的引物和377型DNA序列自动分析仪，检测了1例复发前后GBM所有22对常染色体上多达382个微卫星位点。相邻2个微卫星位点之间的平均距离为10cM。结果：对原发肿瘤标本的等位基因谱分析显示，染色体9p21上D9S157位点和10q21.3-26.3上D10S537、D10S185、D10S192、D10S597、D10S587、D10S217位点存在LOH。对复发肿瘤标本的研究显示，不但9p21和10q21.3-26.3上LOH的范围扩大，而且在其它多条染色体上也出现了LOH（包括1q,7p,7q,21q,20p,20q,10p,19p,19q)。结论：染色体9p和10q可能在该例GBM的发生中起着重要作用。尽管该病例复发前后的病理诊断相同，复发后GBM存在着更广泛的分子遗传学异常改变，可能伴随着更多肿瘤抑制基因的失活。     

THE FREQUENCY AND MECHANISM OF LOSS OF HETEROZYGOSITY ON CHROMOSOME 11q IN BREAST CANCER

Loss of heterozygosity (LOH, allele loss) occurs frequently on the long arm of chromosome 11 in breast cancer. Seventy-one paired tumour/normal DNA samples from breast cancer patients under 50 years old were studied for allele loss at four microsatellite loci on 11q: D11S29 (11q23.3), NCAM (11q22–q23), D11S968 (11qtel), and D11S1313 (11qcen). The maximum frequency of LOH (≈35 per cent) was found at the D11S29 and NCAM loci. This result is consistent with previous studies and the frequency of allele loss is moderate to high compared with the usual baseline of 0–20 per cent. In most of the cases studied, LOH on chromosome 11q could be accounted for by one of two mechanisms. Either chromosomal non-disjunction had occurred, or sequences stretching from the telomere at least as far as NCAM had undergone deletion or mitotic recombination. These results suggest that a putative tumour suppressor gene is most likely to exist near 11q22–q23. There was a very low frequency of microsatellite instability in the tumours. An association was found between lack of progesterone receptor (PgR) expression and LOH at NCAM , suggesting that deletion of sequences on 11q may prevent high levels of PgR expression in some cases.     

Genetic pathways in the evolution of breast ductal carcinoma in situ

The patterns of allelic loss in 28 cases of pure ductal carcinoma in situ (DCIS) and 25 cases of DCIS associated with invasive ductal carcinoma (IDC) were compared, in order to define whether pure DCIS represented an earlier stage than DCIS associated with IDC in the progression of breast carcinoma. To this aim, the polymerase chain reaction (PCR) was performed on microdissected normal and neoplastic breast tissue, formalin-fixed and paraffin-embedded. Fifteen microsatellite markers were examined, on chromosomes 1p, 3p, 7q, 11q, 12p, 13q, 16q and 17q, mostly focused on regions altered in breast cancer. Loss of heterozygosity (LOH) was greater in pure DCIS than in the DCIS component associated with IDC for 11 out of 15 markers. The difference was statistically significant for D13S260 and D17S800 (p=0.008 and p=0.01, respectively). DCIS associated with IDC showed a lesser degree of alteration than the synchronous IDC component for ten out of 15 markers. In contrast, LOH at D11S1816 and D16S318 was lower in pure DCIS than in DCIS associated with IDC and even greater in the IDC component. These results confirm that DCIS is a possible but not an obligate precursor of invasive breast cancer and suggest that pure DCIS and DCIS associated with IDC may be genetically distinct. The evolution from DCIS to IDC may follow multiple pathways and not a linear model.     

中国鲎素和正丁酸钠对人胃腺癌BGC-823细胞形态与超微结构的影响

目的以正丁酸钠为平行对照,比较观察鲎素和癌细胞诱导分化物对人胃癌细胞形态与超微结构变化的影响.方法采用光镜、扫描电镜与透射电镜,观察经2.0 mg/L鲎素、2.0 mmol/L正丁酸钠和1.0 mg/L鲎素＋1.0 mmoL/L正丁酸钠组合处理的人胃腺癌BGC-823细胞.结果光镜观察显示,鲎素和正丁酸钠以及鲎素＋正丁酸钠处理的BGC-823细胞形态均较为一致,细胞体积增大、趋于扁平铺展状态,细胞核质比例减小,细胞核形状较圆整,核仁数量减少.扫描与透射电镜观察可见经鲎素、正丁酸钠及其组合处理的BGC-823细胞均出现细胞表面微绒毛稀少,细胞边缘丝状伪足减少,片状伪足增多;细胞核形态较规则,核内异染色质减少,常染色质增多：细胞质内线粒体增多,结构较为一致,高尔基复合体呈较为典型发达状态,并出现粗面内质网增多和多聚核糖体减少等变化.结论鲎素具有与正丁酸钠等诱导分化物相似的改变人胃癌细胞形态与超微结构恶性表型特征的作用,和与诱导分化物协同加成的诱导分化效果.     

LOH at the sites of the DCC, APC, and TP53 tumor suppressor genes occurs in Barrett's metaplasia and dysplasia adjacent to adenocarcinoma of the esophagus

Barrett's esophagus carries a 30- to 100-fold increased risk of adenocarcinoma, which is thought to develop via a metaplasia-dysplasia-carcinoma progression. A common genetic abnormality detected in Barrett's adenocarcinoma is loss of heterozygosity (LOH) at the sites of known or putative tumor suppressor genes, of which there are at least 9 associated with esophageal adenocarcinoma. The aim of this study was to identify at which histological stage of carcinogenesis LOH at these sites occur. Microdissection of multiple paraffin-embedded tissue blocks from 17 esophagogastrectomy specimens of adenocarcinoma arising in Barrett's esophagus yielded areas of metaplasia, low-, intermediate- and high-grade dysplasia, and carcinoma. LOH analysis of microdissected tissues was performed using a double polymerase chain reaction technique with 11 microsatellite primers shown previously to have LOH in at least 30% of esophageal adenocarcinomas. Identical LOH was detected in premalignant and malignant tissues in 4 of 17 patients, and was located at 5q21-q22 (D5S346 primer), 17p11.1-p12 (TCF2 primer), 17p13.1 (TP53 primer), 18q21.1 (detected in colon cancer tumor suppressor gene [DCC] primer), and 18q23-qter (D18S70 primer). These results suggest that LOH at the sites of the DCC, adenomatous polyposis coli (APC), and TP53 tumor suppressor genes occur before the development of adenocarcinoma in Barrett's esophagus, and so merit further study as potential biomarkers of neoplastic progression in patients with Barrett's esophagus undergoing endoscopic and histological surveillance.     

THE PATTERN OF K-rasMUTATION IN PULMONARY ADENOCARCINOMA DEFINES A NEW PATHWAY OF TUMOUR DEVELOPMENT IN THE HUMAN LUNG

Codon 12 of the K-ras oncogene was screened for mutations in 65 surgically-resected primary pulmonary adenocarcinomas and in 32 tissue foci of alveolar atypical hyperplasia (AAH) by a polymerase chain reaction (PCR)-based method. Mutations in either position 1 or position 2 of codon 12 were detected in 16 tumours (25 per cent). When analysed by site of origin, mutations were seen in 9/26 (35 per cent) parenchymal and in 0/12 bronchial adenocarcinomas (P<0·02). K-ras mutations were seen in five AAH lesions from four patients. DNA sequencing showed that the great majority of mutations in both adenocarcinomas and AAH were G–T transversions. These findings provide support for the classification of pulmonary adenocarcinomas into bronchial and parenchymal subtypes and also provide molecular evidence to support the importance of AAH in the development of parenchymal cancers. © 1997 by John Wiley & Sons, Ltd.     

MALIGNANT PLEOMORPHIC ADENOMA (MALIGNANT MIXED TUMOR) OF THE TRACHEA. Report of a Case

Malignant pleomorphic adenoma arising in the trachea has not been reported in the literature. We report here a case of malignant pleomorphic adenoma (malignant mixed tumor) occurring in the trachea of a 65-year-old woman. The tumor metastasized to the lung and the chest wall 11 years after complete resection of the primary tumor, which was a polypoid submucosal tumor, 1.3 cm in diameter. Light microscopic examination of the primary and metastatic tumors showed the presence of epithelial and stromal elements, consisting of grandular structures, foci of squamous metaplasia and a myxochondroid stroma. Many tumor cells showed myoepithelial cell features by electron microscopy, and immunoreactivity for S-100 protein and GFAP was also seen in many of them. These findings were consistent with those of pleomorphic adenoma. However, the epithelial elements were cytologically atypical with prominent mitotic figures. Infiltration of the tumor cells into the surrounding soft tissue was also seen. No foci of benign pleomorphic adenoma were found in the primary tumor. These findings indicate that this tumor was not a carcinoma ex pleomorphic adenoma, but a true malignant pleomorphic adenoma (true malignant mixed tumor) of the trachea. ACTA PATHOL JPN 38: 1215∼1226, 1988.     

胃腔内促性腺激素释放激素类似物对大鼠胃和十二指肠生长抑素细胞功能的影响

目的　研究胃腔内促性腺激素释放激素类似物 (GnRH A)对大鼠胃和十二指肠生长抑素免疫阳性细胞及血液、胃液中生长抑素含量的影响。　方法　应用免疫组织化学ABC和ELISA方法分别检测大鼠胃和十二指肠生长抑素阳性细胞的密度及血液、胃液中生长抑素的含量。　结果　胃腔注射GnRH类似物后 ,胃壁内单位面积生长抑素阳性细胞数为 2 6 6± 3 893,与对照组 4 8 3± 6 0 19相比具有差异 (P <0 0 1) ,十二指肠单位面积生长抑素阳性细胞数为 5 1 7± 2 2 14 ,与对照组 5 8 5± 4 4 5 4相比具有差异 (P <0 0 1)。血液中生长抑素ELISA检测的A值为 0 15± 0 0 18,与对照组 0 2 7± 0 0 36相比具有差异 (P <0 0 1) ,胃液中A值为 0 32± 0 0 4 9,与对照组 1 0 2 2±0 36 9相比具有差异 (P <0 0 1)。　结论　外分泌的GnRH对大鼠胃和十二指肠中生长抑素的分泌起显著的抑制作用。     

原发性胃癌中19p部分微卫星多态位点杂合性缺失分析

目的 筛选胃癌19p部分微卫星多态位点的杂合性缺失（loss of heterozygosite,LOH)频率,以初步确定19p上与胃癌相关基因连锁最密切的微卫星多态位点。方法 采用聚合酶链反应-单链长度多态（polymerase chain reaction-single strand length polymorophism,PCR-SSLP)-银染法选取19p上9对微卫星多态标记（D19S424,D19S216,D19S406,D19S413。D19S221,D19S226,D19S411,D19S883,D19S886）,对43例原发性胃癌的杂合性缺失情况进行了分析。结果 43例中22例至少在1个位点发生LOH,总缺失率为48.88%,这9个位点的LOH频率分别为29.63%,11.53%,33.33%,8.57%,13.15%,8.00%,6.45%,6.89%,10.71%,在D19S886也同时出现微卫星不稳定性（microsatellite instability,MSI)17.85%。结论 提示19p上的LOH缺失频发区域可能涉及与人类原发性胃癌发生发展相关基因的存在。     

Microsatellite analysis of breast carcinoma and corresponding local recurrences

Local recurrence is a serious complication of breast carcinoma that reduces quality of life and influences prognosis. The aim of this study was to determine whether local recurrences of breast carcinoma are genetically related to the primary tumours. Forty cases of locally recurrent breast carcinomas (median onset: 3.6 years after primary surgery) were analysed: 22 patients had undergone breast-conserving therapy and 18 mastectomy. Eighteen microsatellites on chromosomes 2p, 3p, 5q, 10q, 11p, 11q, 13q, 17q, 17p, 18p were amplified by PCR using fluorescent-labelled primers, automatically detected after polyacrylamide gel electrophoresis and analysed for loss of heterozygosity (LOH) or microsatellite instability (MSI). Follow-up data were available for 39 cases with a median value of 89 months. All LOH and MSI found in the primary tumours were also present in the corresponding recurrences, indicating that they are genetically related to the primary tumours and not secondary malignancies in the same breast. MSI was found in three cases, of which one harboured MSI at more than two loci. The median value of LOH per case was significantly higher in the recurrent (four per case) compared to the primary tumours (two per case; p < 0.001, Mann-Whitney test), reflecting the genotype of tumour progression. Early local recurrence was associated with specific LOH for TP53.15 (p = 0.018, log-rank test) in the primary tumours. LOH on D13S1699 or D17S855 was associated with lymph node metastases (p = 0.024 and p = 0.019, respectively; chi-square test). In addition, tumour grade, lack of oestrogen or progesterone receptor expression, young patient age and early appearance of local recurrence significantly correlated with poor survival. The development of local recurrence despite clear resection margins may result from residual DCIS distant from the invasive carcinoma, homing of circulating tumour cells, or genetically altered, histologically normal breast tissue not immediately adjacent to the invasive carcinoma.     

胃裸区的冠状断层解剖学研究

目的：为给胃底部和食管腹段疾病的现代影像学诊断和外科治疗提供实用的形态学依据。方法：用成人躯干部连续冠状断层标本30例,研究了胃裸区在连续冠状断层上的典型表现及胃裸区内的结构。结果：（1）胃膈韧带右层与小网膜的后层相续,左层与膈脾韧带右层及胃脾韧带后层相续,向下左、右层靠拢而续为胃胰襞左部。在冠状断层上寻找胃裸区的最佳层面在A11（下腔静脉前份层面（和A12（下腔静脉中份层）。（2）胃裸区居胃膈韧带的左、右层之间,其存在率为100％。胃裸区内除有迷走神经后干,血管、淋巴结外,左肾上腺、左肾上极及胰体上份亦可进入其内。结论：（1）胃为腹膜间位器官。（2）国膈韧带很短且两层间的距离较宽,加上胃裸区较大,胃底后壁的活动度较小,在施行胃底和食管腹段手术构解胃底后壁时,对胃裸区内的结构应给与足够的重视。     

胰岛淀粉样多肽在人胎胃的定位研究

目的 探讨胰岛淀粉样多肽(IAPP)在人胎胃的表达和定位。方法 免疫组织化学SABC法对12例11～20周人胎胃体部IAPP免疫反应(IR)细胞进行定位研究。结果 人胎11周,胃黏膜中已有IAPP-IR细胞,主要定位于胃底腺。14～20周,胃黏膜中IAPP-IR细胞数量逐渐增多,免疫反应显色也逐渐加深。经与HE染色的邻片比较,部分IAPP-IR细胞与壁细胞定位一致。结论 IAPP-IR细胞在人胎儿期已存在于胃体部。     

白血病染色体易位断裂点区域微卫星的不稳定性和杂合性丢失

目的探讨白血病染色体微卫星不稳定性（ＭＳＩ）及杂合性丢失（ＬＯＨ）。方法用白血病易位断裂点区域附近的多个微卫星位点检测３０例不同类型白血病基因的不稳定性及杂合性丢失。结果２１例白血病存在ＭＳＩ或者ＬＯＨ。其中１５例急性白血病中１１例示１个以上位点有ＭＳＩ；６例慢性粒细胞性白血病中４例患者在３个以上位点有ＭＳＩ；１５例急性白血病中２例有ＬＯＨ，比其他白血病发生率高。结论白血病和骨髓增生异常综合征均存在较高的基因不稳定性，但这些基因变化与不同类型的白血病的染色体易位断裂点没有特殊联系     

人胚肺成纤维细胞与肺腺癌细胞共同培养时增殖率的变化

用一种新的细胞培养技术，选用人肺腺癌细胞及人胚肺成纤维细胞，将两者这一定顺序定位培养在同一容器内，用原位计数法分别测定两种细胞在１２，３６，６０，８４小时培养的细胞数法计算两种在不同时相的增殖率，观察人肺腺癌细胞和成纤维细胞的相互影响。