卵巢癌血清DNA 3号染色体短臂基因杂合性丢失的研究

目的：探讨血清NDA3号染色体短臂（3p14,25)等位基因杂合性丢失与卵巢癌发生,发展的相关性,方法：采用聚合酶链反应并结合二核苷酸重复序列多态性方法,分别对38例卵巢癌及8例卵巢良性肿瘤患者的血清DNA 3p14,25的4个微卫星位点（D3S1029,D3S1228,D3S1300,D3S1481）杂合性丢失进行检测,另外,检测38例中18例患者的组织及相应血清DNA 3p14,25杂合性丢失。:18列卵巢癌血清与肿瘤组织DNA3p14,25基因2个微卫星位点杂合性丢失率之间存明显的相关性（P＜0．05）,38例血清DNA标本中有29例（76％）至少在1个微卫星位点出现杂合性丢失,17例（45％）有2个以上微卫星位点出现杂合性丢失,卵巢癌II期,Ⅲ期,Ⅳ期出现杂合性丢失率分别为1／4,78％,8／9,。卵巢癌病理类型仅在微卫星D3S1029位点的杂合性丢失率比较,差异有显著性（P＝0．0074）。8例良性肿瘤组织及血清均未出现杂合性丢失。结论：卵巢癌患者血清DNA与肿瘤组织DNA3p14,25出现杂合性丢失密切相关,血清3p14,25出现杂合性丢失率与卵巢癌恶性程度有关。     

Evaluation of a region on chromosome 1p in ovarian serous carcinoma that is frequently deleted in uterine papillary serous carcinoma.

OBJECTIVE: The goal of this study was to assess the involvement of chromosome 1p deletion in ovarian papillary serous carcinoma (OPSC) via high-resolution physical mapping to detect a candidate tumor suppressor gene previously implicated in uterine papillary serous carcinoma (UPSC) tumorigenesis. METHODS: Previous studies have demonstrated a high rate of loss of heterozygosity (LOH) within a critical region of chromosome 1p in UPSC, suggesting the presence of a putative tumor suppressor gene important in the development of UPSC. To compare the genetic changes in OPSC with those in UPSC, seven microsatellite repeat polymorphisms were used to evaluate LOH in primary OPSC specimens. Allelic intensity was compared between normal and tumor DNA from microdissected, formalin-fixed, paraffin-embedded specimens. In addition to the same seven 1p markers used for UPSC, three additional non-1p markers for chromosomes 1q, 11q, and 2p were tested to determine a baseline rate of LOH. RESULTS: Overall, 26.6% (8/30) of OPSC (vs 63.2% of UPSC) were characterized by loss at either of the two loci that define the critical region for UPSC. Rates of LOH at each of the 1p loci in the OPSC tumors ranged from 5.6 to 38.9%, which are compatible with background rates of loss for OPSC. LOH at non-1p loci was 29.2%. CONCLUSION: While a tumor suppressor gene on 1p appears to be an important genetic event in the development of UPSC, a similar rate and pattern of loss on 1p are not identified in OPSC. Thus, despite the striking clinical similarities between UPSC and OPSC, tumorigenesis in these carcinomas appears to occur via distinctly different pathways.     

Loss of heterozygosity analysis in uterine cervical adenocarcinoma

OBJECTIVE: Uterine cervical adenocarcinoma (CAC) is a rare form of cervical cancer, constituting only 5-8% of all cervical epithelial malignancies. Loss of heterozygosity (LOH) analysis of CAC was undertaken to identify alterations of chromosomal loci that may play important roles in the development of this tumor type. METHODS: We analyzed loss of heterozygosity (LOH) using a total of 50 markers on 20 chromosomal arms in 37 cases of microdissected CAC DNA. RESULTS: LOH of >40% was observed on 2p (50%), 3p (45%), 9p (45%), 11q (46%), 17p (57%), 17q (44%), 18q (57%), and 19p (44%). LOH of 30-40% was observed on 6p (38%), 6q (40%), and 10q (31%). Overall, mean LOH was 34% and fractional allelic loss (FAL) was 0.34. High-level and low-level microsatellite instability (MSI) was shown in four cases (11%) and six cases (16%), respectively. Frequency of LOH on10q was significantly higher in endometrioid-type than endocervical-type adenocarcinoma (71% versus 20%; P < 0.05). Conversely, 6q LOH was higher in endocervical type than endometrioid type (0% versus 60%; P < 0.05). 19p13.3 has been reported to be frequently deleted in adenoma malignum, a histological subtype of CAC. To define the critical regions of LOH in CAC in general, we further performed deletion mapping of 19p using 13 markers. Unlike adenoma malignum, multiple regions on 19p appeared to be important loci of LOH for CAC. CONCLUSION: CACs develop with frequent LOH of multiple chromosomal arms, which may be related to its aggressive clinical behavior and poor prognosis. LOH of 10q may be unique to endometrioid-type CAC.     

Cervical intraepithelial neoplasia: prognosis by combined LOH analysis of multiple loci

OBJECTIVE: Cervical intraepithelial neoplasias (CIN) show markedly variable clinical behavior. Clinically, it is important to distinguish CIN lesions with different behaviors and identify those likely to persist and progress. The purpose of this study is to explore whether CIN lesions with different clinical behaviors can be stratified by analysis of loss of heterozygosity (LOH) at multiple loci. METHODS: One hundred sixty-four cases of CIN (54 CIN1, 59 CIN2 and 51 CIN3) were screened for LOH at 12 microsatellite markers including 10 from 3p14, 3p21-22, 6p21 and 11q23. LOH was correlated with clinical follow-up data and high-risk HPV infection. RESULTS: In a pilot study of 71 cases of CIN, screening of 12 microsatellite markers identified four (D3S1300, D3S1260, D11S35, and D11S528) at which LOH was significantly associated with disease persistence/progression. These four markers were further investigated in a larger cohort, which brought the total number of cases examined to 164. Combined analysis of LOH at the above four loci permitted the identification of 22-47% of CIN lesions depending on the histological grade, which showed disease persistence/progression. LOH at these loci was significantly associated with HPV16 infection. Bioinformatic analysis identified several candidate genes including the fragile histidine triad gene and progesterone receptor gene that may be the target of deletions. CONCLUSIONS: LOH at D3S1300, D3S1260, D11S35 and D11S528 was significantly associated with cins that showed persistence/progression, and combined LOH analyses at these loci could be used to identify such cases.     

Frequent allelic imbalance of tumor suppressor gene loci in cervical dysplasia.

In addition to human papillomavirus (HPV) infection, loss of heterozygosity (LOH) at tumor suppressor gene loci has been frequently observed in cervical cancer. Thus, it may be assumed that detection and characterization of specific LOH profiles in preneoplastic lesions, in addition to HPV typing, might facilitate assessment of progression risk of cervical dysplasia. In this study, the type and frequency of allelic imbalance (allelic loss or allelic reduction) were analyzed in 24 unrelated cervical lesions using 14 polymorphic microsatellite markers at different tumor suppressor gene loci. No allelic loss was observed in four condylomatous lesions, whereas 2 of 13 (15%) CIN I lesions displayed allelic loss at 3p25 and 5q11-13. In high-grade lesions, however, allelic loss occurred in four of six (66%) cases at multiple chromosomal regions (3p14-25, 5p15, 5q11, 5q21, 11p15, and 17q21). Allelic reduction was observed in 4 of 13 (30%) low-grade lesions and 3 of 6 (50%) high-grade lesions. LOH was confined to lesions infected by high-risk HPV types. These data suggest that chromosomal instability is an early event in cervical carcinogenesis. The detection of LOH on multiple chromosome 3p loci in 50% of high-grade lesions suggests that a specific marker panel encompassing this region might enable better assessment of which lesions are likely to regress, persist, or progress.     

LOH of chromosome 6q compared with LOH of 17q and 18q in ovarian cancers: relationship to p53 expression and clinicopathological findings

In 41 ovarian epithelial tumors (7 borderline and 34 invasive), loss of heterozygosity (LOH) of chromosomes 6q, 17q, and 18q was examined using 4 microsatellite markers: ER (6q 25–1), BRCA1 (17q21), DCC (18q21), and D18S58 (18q23). The LOH was compared with clinicopathological findings, including p53 and ER expression. In borderline tumors, LOH and p53 expression were never found, while in invasive carcinomas LOH and p53 were found in 71% and 59% of cases, respectively. In particular, in invasive carcinomas 6q LOH represented a marker distinguishing two groups of tumors; those with 6q LOH were only of serous histotype and at advanced stages (III/IV). No significant difference was found for any of genes in 5-year survival of the patients. No correlation was found between ER expression and ER LOH, as well as between biological aggressiveness and 17q and/or 18q LOH.
We conclude that p53 and LOH of the investigated loci distinguish borderline from invasive ovarian carcinomas; moreover, the comparison of these results with clinicopathological parameters suggests that the presence of 6q LOH may be a factor accounting for greater biologic aggressiveness independent of the histologic subtype.     

Epigenetic and genetic alternation of PTEN in cervical neoplasm

OBJECTIVES: We studied the role of epigenetic and genetic changes of PTEN in the development of squamous cell carcinoma (SCC) of the uterine cervix and their value as a prognostic factor. METHODS: Ten high-grade cervical intraepithelial neoplasia (CIN-H) and 62 SCC tissues were used in this study. Microdissection was performed before loss of PTEN function through methylation of promoter CpG islands, deletion and mutation were studied. The findings were verified with PTEN protein expression and correlated with clinicopathologic information. RESULTS: PTEN mutation assessed by single-strand conformation polymorphism (PCR-SSCP) was not noted in any of the 62 SCC. Loss of heterozygosity (LOH) was only seen in eight SCC. PTEN promoter methylation was detected in 40% (4/10) of CIN-H and 58% (36/62) of SCC specimens. Loss of PTEN protein expression was associated with methylation of PTEN. PTEN methylation was not related to patient age, tumor grade or stage. Patients with persistent disease or who died of disease had a significantly higher percentage of PTEN methylation than those without evidence of recurrence. Multivariate Cox regression models confirmed PTEN was an important significant predictor both for total and disease-free survival after controlling age, pathologic grade and clinical stage. CONCLUSIONS: PTEN methylation and loss of PTEN expression are early events in the development of cervical cancer and may have prognostic significance.     

Expression of p27, p21, and p16 protein in early squamous cervical cancer and its relation to prognosis

OBJECTIVE: To examine the prognostic significance of the protein expression of the cyclin-dependent kinase (cdk) inhibitors p27, p21, and p16 in early squamous cervical cancer (SCC). METHODS: From 221 [corrected] patients who underwent radical hysterectomy and bilateral lymphadenectomy at our institution for stage IB SCC between 1987 and 1993, tissue blocks were available for this study. Immunohistochemistry using monoclonal antibodies against p27, p21, and p16 was used to examine protein expression. Ten patients who underwent hysterectomy for uterine prolaps served as controls. RESULTS: p21 and p16 expression were increased in SCC and high expression was observed in 20% (44/221) and 43% (94/220) of tumors, respectively. p27 was decreased in SCC and low expression was found in 80% (177/221) of tumors. In univariate analysis all classical clinicopathological parameters were associated with prognosis. Low p16 expression was significantly related to decreased overall (P = 0.036) but not disease-free survival (P = 0.103). In multivariate analysis, deep stromal invasion but none of the cdk inhibitors was independently related to survival. CONCLUSION: The cdk inhibitors p27, p21, and p16 are not independently associated with prognosis in stage IB SCC.     

Frequent occurrence of loss of heterozygosity among tumor suppressor genes in uterine leiomyosarcoma

OBJECTIVE: Leiomyosarcoma of the uterus is a rare smooth muscle tumor; it is extremely malignant and the rates of local recurrence and metastasis are high. Since tumor suppressor genes are commonly altered in malignant tumors, it is possible that mutations in such genes are involved in the development of uterine leiomyosarcoma. METHODS: Fifty-five patients (37-70 years of age) diagnosed as having smooth muscle tumors of the uterus were selected. DNA was extracted from four or five 8-microm-thick consecutive tissue sections of each smooth muscle tumor from the paraffin-embedded blocks. Loss of heterozygosity (LOH) was investigated at nine loci within or close to tumor suppressor genes (TP53, RB1, DCC, NM23, WT1, D14S267, P16, DPC4, PTCH). RESULTS: Nineteen of twenty leiomyosarcomas revealed at least one instance of LOH among eight of the nine markers tested (one locus showed no LOH at all). In fact, 11 of the 20 cases exhibited two or more instances of LOH and, of the remaining 9 cases, 4 showed a point mutation of p53 in addition to an alteration in one of the 9 markers, while one exhibited a p53 mutation only. CONCLUSION: An accumulation of genetic alterations among tumor suppressor genes may play a key role in the tumorigenesis and progression of uterine leiomyosarcoma.     

X chromosomal and autosomal loss of heterozygosity and microsatellite instability in human cervical carcinoma

The study analyzes tumor material and normal tissue from 27 patients with pure squamous cell carcinoma of the uterine cervix for loss of heterozygosity (LOH) and microsatellite instability (MSI) on 14 autosomal and 11 X chromosomal loci. Overall, 4-40% of the informative cases showed LOH at autosomal regions with the highest frequency at 3p (21-40%) and a marked frequency at 2q35-q37.1 (12.5%) and 17p13.3 (10%), representing regions with putative tumor suppressor gene (TSG) function. The frequency of X chromosomal LOH ranged from 4% to 20%, with a maximum at Xq28 (20%) and Xq11.2-q12 (17%), again indicating alterations in TSG. A 12% LOH was seen at Xq21.33-q22.3, a region encoding a protein with a regulatory function in the cell cycle via cyclin-dependent kinases. MSI was detected in autosomal regions in up to 7% in regions linked to the X chromosome in up to 11%, probably indicating alterations of mismatch repair mechanisms. Our results and those obtained from the literature suggest that autosomal LOH and MSI in carcinomas of the cervix uteri are predominantly found at regions with putative TSG function. Beside TSG alterations, X chromosomal LOH is probably more strongly connected to disturbances in cell cycle regulation.     

Microsatellite analysis in serous tumors of the ovary.

Thirty-four serous ovarian neoplasms were analyzed for microsatellite instability (MIN) and loss of heterozygosity (LOH) at 20 chromosomal loci of eight different chromosomes, including the map positions of the six known mismatch repair genes. Twelve of the 20 (60%) serous ovarian tumors of low malignant potential (LMP) and 13 of 15 (87%) samples of serous ovarian carcinomas, taken from 14 patients, showed LOH at one or more of the analyzed microsatellite loci. In serous carcinomas, LOH of the dinucleotide repeat D7S521 was most frequent. Six of 20 (30%) LMP tumors were also affected by MIN at more than one locus, whereas in the carcinomas MIN was found only sporadically (p < 0.025). No correlation between increased occurrence of MIN and LOH at the chromosomal loci of the known mismatch repair genes were discovered for these LMP tumors. Although our observation regarding LOH frequency in serous LMP tumors and serous carcinomas is compatible with the hypothesis that serous LMP tumors might represent precursor lesions of invasive carcinomas, the results concerning the occurrence of MIN suggest that the mechanisms of tumorigenesis of some tumors of LMP are distinct from those in invasive serous carcinomas.     

Loss of heterozygosity alterations associated with progesterone therapy in endometrial hyperplasia and adenocarcinoma

Loss of heterozygosity (LOH) was analyzed in four patients with endometrial hyperplasia (EH) with atypia (two patients) and without atypia (two patients) and in five patients with endometrial adenocarcinoma (EAC) to clarify the clinicopathologic relationship between genetic alterations and hormone therapy. Each patient was initially administered high-dose medroxyprogesterone acetate (MPA) as a uterine-sparing treatment. The five microsatellite markers used to analyze LOH were at chromosomal loci 8p22.1, 8p21, 8p21.3, 8p22, and 8p22. DNA was extracted from paraffin-embedded sections before, during, and after MPA therapy using laser capture microdissection. As a result, LOH was more frequently detected after MPA therapy (overall ratios were 16, 17, and 29% before, during, and after MPA therapy, respectively). LOH is more easily detected in EH loci than in EAC loci before MPA. For EAC, initial LOH detection on chromosome 8 may be related to an incomplete response to MPA, but negative LOH does not guarantee a favorable treatment outcome. For EH or atypical endometrial hyperplasia, it is unknown whether LOH alteration associated with MPA therapy is related to atypia of the disease.     

Allelic Losses at Chromosome 3p Are Seen in Human Papilloma Virus 16 Associated Transitional Cell Carcinoma of the Cervix

OBJECTIVE: Transitional cell carcinomas (TCCs) of the cervix are rare neoplasms of the female genital tract. Although these tumors display urothelial differentiation, there is controversy regarding their histogenetic relationship to squamous cell carcinomas (SCC) of the cervix versus transitional cell carcinomas of the bladder. METHODS: We performed partial allelotyping of five TCCs of the cervix using 23 polymorphic markers located on chromosomes 3p and 9, which demonstrate frequent and early losses in cervical SCC and urothelial TCC, respectively. Multiplex polymerase chain reaction was used on DNA extracted from archival paraffin-embedded tissue using precise microdissection. Additionally, P53 gene mutation analysis was performed using single-strand confirmation polymorphism (SSCP) and the presence of human papilloma virus (HPV) sequences was analyzed using general and specific (types 16 and 18) primers. RESULTS: General HPV sequences were demonstrated in all cases, but the oncogenic strain HPV 16 was present in only three (60%) of the five tumors; no HPV 18 was detected in any sample. Three of five TCCs, all harboring HPV 16 sequences, demonstrated concurrent allelic losses at several 3p loci (specifically 3p12, 3p14.2 [the FHIT gene locus], 3p21.3, and 3p22-24.2). LOH at a single locus on 9q32-qter was demonstrated in one tumor; no other deletions were seen on chromosome 9. P53 gene mutations in exons 5-8 were absent by SSCP analysis. CONCLUSIONS: The infrequent involvement of chromosome 9 in TCCs of the cervix, along with the concurrent presence of 3p LOH and oncogenic HPV 16 in a subset of tumors, suggests a closer histogenetic relationship of this neoplasm to cervical SCCs rather than urothelial TCCs.     

Squamous Cell Carcinoma Arising in Mature Cystic Teratoma of the Ovary

Treatment results of 26 patients with squamous cell carcinoma (SCC) arising in mature cystic teratoma of the ovary were analyzed. Four nulliparous patients with stage Ia tumors underwent conservative salpingo-oophorectomy. Following surgery, 2 patients had successful pregnancies. The remaining 7 patients with stage Ia tumors were observed after hysterectomy and bilateral salpingo-oophorectomy. Fifteen patients with stage Ic–IV tumors underwent cytoreductive surgery followed bycis-platinum-based chemotherapy with or without sequential radiotherapy. The mean survival was 63.9 months. The overall actuarial disease-free survival at 2 years was 69%, and by stage was as follows: stage I, 100% (13/13); stage II, 100% (2/2); stage III, 30% (3/10); and stage IV, 0% (0/1). A significant difference in disease-free survival was noted in stage (P= 0.0001). Optimal versus suboptimal operation was associated with a median Kaplan–Meier survival of 65 months versus 34.8 months, with actuarial disease-free survival at 2 years of 60 and 0%, respectively (P= 0.0210). Our study shows that 67% (16/24) of the patients had SCC antigen levels exceeding 2 ng/ml, which by stage was as follows: stage I, 5/11 (45%); stage II, 1/2 (50%); stage III, 9/10 (90%); and stage IV, 1/1 (100%). After completion of treatment, all 8 patients with recurrent lesions had reelevated SCC antigen levels in series SCC antigen monitoring. In conclusion, positive prognostic factors of disease-free survival were optimal cytoreduction and lower FIGO stage. We suggest that multimodality therapy, including aggressive cytoreduction followed bycis-platinum-based chemotherapy with or without sequential radiotherapy, is recommended. In addition, we suggest that serum SCC antigen monitoring may be helpful in early detection of cancer recurrence.     

Allelotype of papillary serous peritoneal carcinomas.

OBJECTIVE: Papillary serous peritoneal carcinoma (PSPC) is histologically indistinguishable from papillary serous ovarian carcinoma (PSOC) with a similar clinical presentation, yet may differ in its carcinogenesis. The purpose of this study was to determine the incidence of allelic loss and the frequency of p53 mutation by p53 overexpression in PSPC compared to PSOC. METHODS: An allelotype analysis of 26 patients with PSPC was performed using 39 microsatellite markers from 25 chromosomal arms. Thirty-seven previously studied patients with PSOC served as the comparison. P53 mutations were detected by immunohistochemical protein overexpression. RESULTS: There was significantly less LOH in PSPC than PSOC. Both the number of chromosomes with LOH and the proportion of tumors with allelic loss were less frequent. Significant LOH, defined as >/=30% of informative tumors having loss at a chromosome locus, was seen on 4 chromosome arms in PSPC: 12p, 17p, 17q, and 18q, compared to 18 arms in PSOC: 4q, 5q, 6p, 6q, 9p, 9q, 12p, 12q, 13q, 15q, 16q, 17p, 17q, 18q, 19p, 19q, 22q, and Xq (P < 0.001). The median LOH frequency was higher in PSOC than PSPC, 43% versus 33%, respectively (P = 0.013), and more PSOC tumors had LOH than PSPC tumors, 91% versus 65% (P = 0.042). P53 overexpression was detected in 80% of PSPC tumors. CONCLUSIONS: LOH occurs less frequently in PSPC compared to PSOC. Chromosomal regions with high frequencies of LOH common to PSPC and PSOC, such as 12p, 17p, 17q, and 18q, may harbor tumor suppressor genes important in the carcinogenesis of both malignancies and likely include p53.     

Investigation of loss of heterozygosity at specific loci on chromosomes 3p, 6q, 11p, 17p and 17q in ovarian cancer

In an attempt to further define the genetic events in the pathogenesis and progression of human ovarian cancer, an analysis of constitutional and ovarian carcinoma DNA samples revealed loss of heterozygosity (LOH) at specific loci on chromosomes 3p (38%), 6q (23%), 11p (33%), 17p (82%) and 17q (62%). In contrast, LOH was not observed in benign or borderline tumors. No significant association could be demonstrated between LOH at the loci studied and tumor stage, histologic subtype, grade or patient outcome. Further analyses of large tumor panels are now required to determine the relationship between LOH at these loci and the clinicopathological behavior of ovarian tumors.     

遗传性卵巢癌组织中BRCA1基因杂合性丢失的研究

目的：探讨乳腺癌基因（ＢＲＣＡ１）在遗传性卵巢癌发生、发展中的作用,并分析ＢＲＣＡ１ 基因杂合性丢失（ＬＯＨ）。方法：应用多聚合酶链反应技术（ＰＣＲＴ）,对１０份遗传性卵巢癌组织中ＢＲＣＡ１ 基因内部的Ｄ１７Ｓ８５５ 微卫星位点进行ＬＯＨ检测。结果：２份发生杂合性丢失,ＬＯＨ发现率为２０％ 。结论：ＢＲＣＡ１在遗传性卵巢癌中具有肿瘤抑制基因的作用,同时不能排除其他肿瘤抑制基因的作用。     

Distinct subtypes of serous ovarian carcinoma identified by p53 determination

OBJECTIVE: The overall prognosis of ovarian carcinoma is poor. However, the outcome of apparently similar cases is highly variable, and molecular markers that would predict disease outcome in a clinically useful manner are lacking. We investigated the value of p53 expression as a disease determinant in serous carcinoma, which is the most common type of ovarian carcinoma and has shown the highest frequency of p53 alterations. METHODS: Tissue microarray constructed of 522 serous ovarian carcinomas was examined immunohistochemically using DO-7 monoclonal antibody against p53 protein. The findings were correlated with overall and disease-free survival, response to therapy, and clinicopathological characteristics of the patients. RESULTS: Both excessive and completely negative p53 staining confered poor patient outcome and were considered aberrant p53 expression. Patients with aberrant p53 (59% of the carcinomas) showed 5-year overall survival of 26% (20-31%), whereas patients with normal p53 expression (41% of the carcinomas) showed 5-year overall survival of 79% (95% CI, 74-85%) (P < 0.0001). The association of aberrant p53 with poor prognosis was independent of clinicopathological parameters, e.g., stage and grade. In addition, aberrant p53 status was significantly associated with shorter disease-free survival (P < 0.0001) and poor response to therapy (P < 0.0001). In the most common subgroups, stage III and stage I carcinomas, 5-year overall survival rates for patients showing normal p53 vs. aberrant p53 were 72% vs. 19% (P < 0.0001) and 99% vs. 56% (P < 0.0001), respectively. CONCLUSION: P53 expression status divides serous ovarian carcinomas into two distinct subtypes: one with a relatively good prognosis and the other with a particularly poor outcome.     

Leiomyosarcoma of the uterus: a clinicopathologic study of 21 cases

BACKGROUND: The purpose of the present study was to identify the prognostic factors of and to determine the most appropriate mode of treatment for uterine leiomyosarcoma. METHODS: We reviewed the hospital records, including surgical notes and pathologic reports, of 21 patients with uterine leiomyosarcoma treated at Chang Gung Memorial Hospital, Kaohsiung, Taiwan, between 1987 and 1997. Univariate analysis was performed using the log-rank test. Cox regression was used to identify independent prognostic factors. RESULTS: The mean follow-up time was 30 months, and the 5-year disease-free survival rate was 55%. Evaluating the correlation between clinicopathologic parameters and survival, early stage (p = 0.0002), tumor cells without necrosis (p = 0.0026), low-grade tumor (p = 0.015), absence of vascular space involvement (p = 0.006), and tumor without atypia (p = 0.016) were associated with good prognosis. However, in a multivariate analysis using the Cox model, only advanced stage (p = 0.032) and tumor necrosis (p = 0.032) were found to be independent poor prognostic factors. The 5-year disease-free survival was only 21% in patients with the presence of any one or both of these two factors. Five of 11 patients in this group had received aggressive adjuvant therapy after surgery, and none of them survived more than 19 months. CONCLUSIONS: We found that patients with advanced stage or presence of tumor necrosis had an extremely poor prognosis. Adjuvant therapy seemed to play a limited role, and provided no survival benefit. Treatment for these patients should be palliative until effective therapeutic modalities prove otherwise.     

Allelic imbalances in endometrial stromal neoplasms: frequent genetic alterations in the nontumorous normal-appearing endometrial and myometrial tissues

OBJECTIVES: Endometrial stromal sarcoma (ESS) is among the rarest primary malignant tumors of the uterus. The aim of this study was to examine the possibility of loss of heterozygosity (LOH) and microsatellite instability (MIS) in different tissue components of ESS. METHODS AND MATERIALS: Using PCR, we examined DNA extracts from microdissected tissues of 27 uterus samples containing malignant stromal cells of ESS (20 low grade and 3 high grade sarcomas), benign tumor cells of endometrial stromal nodules (ESN, 4 cases) as well as tumor-free myometrial and endometrial tissues close to and distant from the tumors. Normal cervical tissues (epithelial cells, stroma cells) were also microdissected and analyzed. Fifteen polymorphic DNA markers (chromosomes 2p, 3p, 5q, 10q, 11q, 13q, and 17p) were tested to identify possible genetic alterations. Samples from 10 women with prolapsed uteri without any histopathologic abnormalities were also selected as controls. RESULTS: While no genetic alterations could be identified in 12 (44.5%) ESS cases, 15 (55.5%) revealed LOH with at least one polymorphic DNA marker. LOH were found in 3 (100%) high-grade sarcomas, 10 (50%) low-grade ESS, and 2 (50%) benign ESN. Although LOH was found more often in the neoplastic stromal cells, several cases showed concurrent and independent LOH in the tumor-free myometrial or endometrial tissues either close to or distant from the tumors. The most common genetic abnormality (LOH) was observed at PTEN, a tumor suppressor gene located on chromosome 10q. No tumor was associated with microsatellite instability (MSI). The control group without any histologic abnormalities did not show LOH or MSI. CONCLUSIONS: The frequent occurrence of LOH and the lack of MSI suggest that loss of function(s) of tumor suppressor genes and not mismatch repair deficiency plays a key role in the pathogenesis of endometrial stromal neoplasms. The concurrent and independent occurrence of LOH in the stromal tumor cells and the tumor-free and normal-appearing myometrial and endometrial tissues strongly support the concept of genetic alterations in microenvironmental tissues and the interaction(s) between different tissue components in the development and progression of endometrial stromal neoplasms.