Clinical and pathologic significance of microsatellite instability in endometrial cancer

Abstract. Wong YF, Ip TY, Chung TKH, Cheung TH, Hampton GM, Wang VW, Chang AMZ. Clincal and pathologic significance of microsatellite instability in endometrial cancer.
Microsatellite instability (MSI) is identified as electrophoretic shifts in allele sizes of microsatellite DNA sequences. It is characteristic of a subset of sporadic colorectal tumors as well as hereditary nonpolyposis colorectal cancer (HNPCC). The cells that display MSI are thought to be susceptible to increased mutability. MSI has been detected in a wide variety of human tumors, but the influence of this form of genetic instability on disease initiation and progression remains unclear. Using a polymerase chain reaction (PCR)-based method we screened 50 sporadic primary endometrial carcinomas to characterize the prevalence of MSI in these tumors and analyze the correlation of MSI with clinicopathologic parameters in this malignancy. Fifteen cases (30%) displayed low frequency of MSI (MSI-L) showing MSI at one locus in 5 loci examined. Two cases (4%) showed high frequency of MSI (MSI-H) having MSI at 2 or more loci. Taking MSI-L and MSI-H cases together as MSI-positive, statistical analysis of patient age, tumor grade, and stage failed to disclose significant differences or trends between cases with MSI-positive and MSI-negative ( P > 0.05). No significant relationship was observed between patients with MSI and without MSI ( P > 0.05), however, the MSI exhibited only in those cases without evidence of disease at the 24th month after treatment. The difference is statistically significant when compared with patients who are alive with disease or died of disease ( P < 0.01). However, the overall survival curves were not statistically different. We conclude that MSI is present in a subgroup of endometrial cancer.     

Clinicopathologic and immunohistochemical features and microsatellite status of endometrial cancer of the uterine isthmus.

To clarify the clinicopathologic, molecular, and immunohistochemical characteristics of uterine isthmic endometrial cancer (UIE), we examined 13 cases of UIE and compared them with 33 cases of endometrial cancer of the uterine corpus (UCE) with respect to clinicopathologic factors, the expression of p53, the estrogen receptor (ER) and the progesterone receptor (PR) status, DNA ploidy, and microsatellite instability (MSI). Five (38.4%) of the UIE patients had stage I, two (15.4%) had stage II, and six (46.2%) had stage III disease (FIGO 1988). Myometrial invasion was confirmed in 92.3% of the UIE patients, and these patients had a higher (p < 0.05) frequency of > 50% myometrial invasion (46.2%) than the patients with UCE (15.2%). Moreover, the UIE patients had a higher frequency of positive peritoneal cytology (p < 0.05) and pelvic lymph node metastases (p < 0.05). No UIE tumors exhibited MSI, and the tumors in these patients had a higher expression of p53 (p < 0.01), a lower expression of ER (p < 0.05) and PR (p < 0.05), and a higher frequency of DNA aneuploidy (p < 0.01) than the UCE tumors. These findings suggest that the UIE is clearly different from UCE in the clinicopathologic, immunohistochemical features, and microsatellite status.     

Body mass index: relationship to clinical, pathologic and features of microsatellite instability in endometrial cancer

OBJECTIVES: There is a well known association between obesity and endometrial cancer. We sought to examine the relationships between body mass index (BMI), as a measure of obesity, and known demographic, clinical, and molecular characteristics of microsatellite instability and MLH1 promoter methylation in a cohort of patients with endometrial cancer. METHODS: Corpus cancer specimens were prospectively obtained from 473 consecutively enrolled patients between 1992 and 2004. Clinical and pathologic data were extracted from review of the medical record. Microsatellite instability (MSI) was evaluated in all tumors, and methylation of the MLH1 promoter was determined for MSI positive tumors. RESULTS: The median (SD) age and BMI were 64.8 years (11.9) and 33.5 (9.4), respectively. Histology included 376 endometrioid (79%), 69 serous/clear cell or mixed (15%), and 28 sarcomas (6%). Median BMI was 32.4 for endometrioid, 31.0 for serous/clear cell or mixed, and 27.8 for sarcomas (p=0.14). BMI was negatively associated with age at surgery (p<0.01). The remainder of analyses excluded sarcoma histology. BMI was associated with stage of disease; patients with stage I/II disease had significantly higher BMI than those with stage III/IV disease (32.6 vs. 30.6; p=0.02). In relation to molecular features of endometrial cancer, BMI was significantly different between MSI positive tumors compared to MSI negative tumors (30.3 vs. 32.7; p=0.02). MSI was also significantly different between tumor histology, occurring with a higher frequency in Type I than Type II tumors (p<0.01). CONCLUSIONS: The majority of endometrial cancer patients are obese. Those with higher BMI are more likely to be younger, present with early stage disease, and have MSI negative tumors.     

Low-risk endometrial carcinoma: assessment of a treatment policy based on tumor ploidy and identification of additional prognostic indicators.

OBJECTIVES: The aims of this study were (1) to assess a treatment policy for patients with low-risk endometrioid endometrial carcinoma where adjuvant treatment decisions have been based on ploidy status of the tumor, and (2) to screen diploid, low-risk tumors for additional features of prognostic significance. METHODS: Between 01/1992 and 08/1996, 406 patients were referred to the B.C. Cancer Agency-Vancouver Clinic with typical endometrial adenocarcinomas limited to <50% myometrial invasion and no vascular space invasion or grade 3 disease on pathology review ("low-risk stage I endometrial carcinoma"). Patients were prospectively assigned to treatment groups based on tumor ploidy. Those patients with aneuploid tumors (n = 91) were treated with adjuvant vaginal vault radiotherapy while those with diploid tumors (n = 315) were followed and treated only at relapse. The hysterectomy specimens from all 14 patients in the untreated, diploid group who relapsed, as well as 28 stage- and grade-matched diploid controls who did not fail, were analyzed by immunohistochemical staining for estrogen receptor (ER), Bcl-2, and p53 proteins. RESULTS: There were no significant differences in the stage (Ia vs Ib) and grade (G1 vs G2) distribution for the diploid and aneuploid groups. Overall median age was 64 years (range 27-90 years) and was also not significantly different for the two groups. The median follow-up for the entire cohort is 45 months (range 1-76 months). There have been 14 failures in the diploid group and 4 failures in the aneuploid group with actuarial 5-year disease-free survival rates of 95.0 and 95.2%, respectively (P = NS). Eight of the failures in the diploid group occurred at the vaginal vault and were all subsequently salvaged with radiotherapy. All but 1 of the failures in the aneuploid group were considered incurable. Of the 14 diploid tumors from patients who failed, 7 stained positively for p53, compared to 4 of 28 diploid controls (P = 0.02). No significant differences were seen in the diploid tumors that recurred, compared to controls, with respect to Bcl-2 or ER expression. CONCLUSIONS: Patients with diploid, low-risk stage I endometrial cancers have excellent prospects for relapse-free and overall survival. Patients with aneuploid tumors treated with adjuvant radiotherapy have the same risk of relapse as untreated patients with diploid tumors; however, their ultimate survival may be lower as a smaller proportion of aneuploid failures are salvageable. While p53 expression in diploid tumors is associated with increased risk of relapse, Bcl-2 and ER are not useful prognostic indicators in this setting.     

Microsatellite instability, expression of hMSH2 and hMLH1 and HPV infection in cervical cancer and their clinico-pathological association

Infection with specific genotypes of human papillomavirus (HPV) has been strongly implicated in cervical carcinogenesis. However, HPV infection alone is insufficient for malignant transformation of the cervical epithelium. An alteration of microsatellite repeats is the result of slippage owing to strand misalignment during DNA replication and is referred to as microsatellite instability (MSI). These defects in DNA repair pathways have been related to human carcinogenesis; however, the role of MSI in the tumorigenesis of cervical cancer remains unclear. The clinical and pathological features of cervical cancers which are MSI-positive have also not been fully characterized. This study investigated the prevalence of MSI in cervical cancer and its relationship to clinico-pathological characteristics and HPV infection. Polymerase chain reaction-based microsatellite assay combined with tissue microdissection was used to examine for MSI in 50 cervical squamous cell carcinomas in Hong Kong women. In addition, the immunohistochemical staining was performed to determine the expression of major DNA mismatch repair genes, hMSH2 and hMLH1. Six cases (12%) displayed a low frequency of MSI (MSL-L) showing MSI at one locus only in 5 loci examined. Seven cases (14%) showed a high frequency of MSI (MSI-H) having MSI at 2 or more loci. Grouping MSI-L and MSI-H cases together as MSI-positive, statistical analysis of HPV infection, tumor grade, clinical stage and clinical status failed to disclose differences between MSI-positive and MSI-negative cases (p > 0.05). However, MSI-H correlated with advanced stage of disease (p < 0.05). Individuals with MSI-H tumors appeared to have reduced overall survival compared to individuals with MSI-L and MSI-negative tumors, but the difference was not statistically significant (p = 0.059). An absence of either MSH2 or MLH1 expression was observed in 2 MSI-L and 4 MSI-H cases, respectively. The results suggest that MSI is present in a subgroup of cervical squamous cell carcinomas, and defects resulting in MSI may be related to tumor progression and possibly poor prognosis in cervical cancer.     

The detection of microsatellite instability in blind endometrial samples—a potential novel screening tool for endometrial cancer in women from hereditary nonpolyposis colorectal cancer families?

Microsatellite instability (MSI) is the phenotypic molecular characteristic of the majority of tumors associated with the hereditary nonpolyposis colorectal cancer syndrome (HNPCC). Women in this group have an increased risk of endometrial cancer (EC). This study aimed to determine whether MSI could be demonstrated in blind endometrial samples from women with EC, HNPCC kindreds undergoing screening for EC, and women with normal endometrium. Twenty-four women with EC, 20 women from HNPCC kindreds, and 20 women undergoing gynecological surgery for benign indications underwent blind sampling. MSI analysis was performed by conventional polymerase chain reaction using fluorescent-labeled primers and automated analysis. Twelve microsatellites were studied with MSI defined as evident when novel alleles were seen in endometrial biopsy samples compared to genomic DNA. Of the 24 EC samples obtained, sufficient DNA for analysis was extracted in 17 cases. Three cases had evidence of MSI in at least 7/12 loci. None of the endometrium from the two other study groups revealed evidence of MSI. This is the first demonstration of MSI in blind endometrial biopsies. The ability to demonstrate MSI in heterogeneous endometrial samples suggests potential for the development of a novel EC screening tool for women in HNPCC kindreds.     

Flow cytometric and clinicopathologic study of complete hydatidiform moles with special reference to the significance of cytometric aneuploidy

OBJECTIVE: As complete hydatidiform moles (CMs) have been studied less with respect to aneuploidy and its clinical implications, the significance of cytometric aneuploidy in CMs was evaluated. METHODS: Two hundred thirty-nine CMs were studied clinicopathologically and analyzed by flow cytometry using formalin-fixed paraffin-embedded tissues. RESULTS: Of 239 CMs, 182 were diploid, 30 were tetraploid, and 27 were aneuploid (nontriploid/tetraploid aneuploid). There were no significant histologic differences among the diploid, tetraploid, and aneuploid CMs. Persistent disease developed in 20 of 114 CMs (17.6%) (16 of 77 diploid, 4 of 18 tetraploid, and none of 19 aneuploid CMs). Eight diploid and three tetraploid CMs were invasive, and one patient each with diploid CM and tetraploid CM developed choriocarcinoma and none of 19 patients with aneuploid CMs had sequelae. CONCLUSION: These results suggest that aneuploid CMs are associated with less risk for persistent disease than diploid or tetraploid CMs. DNA ploidy status may be an independent predictor of persistent disease.     

DNA ploidy and results of first line chemotherapy in ovarian cancer patients

AIM OF THE STUDY: The prognostic significance of DNA ploidy in ovarian cancer patients determined by flow cytometric analysis, in correlation with effectiveness of first line chemotherapy. MATERIAL AND METHODS: DNA ploidy by FC was investigated in group of 102 ovarian cancer patients from fresh frozen samples (4 patients was excluded from the study). RESULTS: Positive answer for first line treatment we notified in 64(62,75%) cases, lack of answer 34 (37,25%) patients. Aneuploidy was more frequent in negative group 31(91,18%), diploid tumours occurred in 3(8.82%) cases. In positive group aneuploid tumours occurred in 29(45,31%) and diploid in 35(54,69%) patients (p<0,001). Median survival in positive group--45 months, in negative group 12 months (p<0,0001). In positive group median survival in patients with aneuploid tumours--31 months, in patients with diploid tumours median survival was not reached (p=0,0102). In negative group DNA ploidy has no impact on survival (p=0,1027) CONCLUSIONS: 1. DNA ploidy determined by flow cytometry is prognostic factor in ovarian cancer patients who answered positively for first line treatment. 2. Aneuploid tumours appear much often then diploid in group of patient who did not answer for first line chemotherapy 3. Patients with diploid tumours have better prognosis. 4. Lack of positive answer for first line treatment is bad prognostic factor.     

The changes in nuclear DNA amounts in the development and progression of uterine cervical cancer determined by cytofluorometry

After microsopically-directed sampling of the tissues from various histological sections taken from 16 cases of cervical dysplasia, 6 cases of carcinoma in situ, 11 cases of stage Ib invasive cervical cancer of keratinizing type, 28 cases of large cell non-keratinizing type and 14 cases of small cell type, nuclear DNA levels of the cells dispersed from the tissues were measured by cytofluorometry after Feulgen stain. The DNA levels of cells obtained from normal cervical squamous epithelium and squamous metaplastic epithelium were in 2C(diploid)-4C(tetraploid) regions and those from mild and severe dysplasia were in 2C approximately 4C or high 4C and in low 2C or 2C approximately high 4C or 8C(octaploid) regions respectively with the mode of 2C. In five of 6 cases (83.3%) of carcinoma in situ, the amount of DNA in the neoplastic cells ranged up to the hyperoctaploid region with the mode of 2C or 4C. There were hyperoctaploid cells in 90.9% of cases of the keratinizing type, 96.4% of cases of the large cell non-keratinizing type and 100% of cases of the small cell type. The incidence of hyperoctaploid cells in samples from superficial invasion (stromal invasion less than 3mm) was not different from that of deep invasion (stromal invasion of more than 5mm) in each histological type. When the modal values for nuclear DNA in the superficially invasive lesions were compared with those of the deeply invasive lesions, aneuploidy was more frequently observed in the lesions of deep stromal invasion, irrespective of the histological type. The dominant changes in the mode according to the depth of stromal invasion were 2C to aneuploid in keratinizing type and 4C to aneuploid in cases of large cell non-keratinizing type and small cell type. The results suggest that the hyperoctaploid and aneuploid cells are useful markers for quantitative discrimination among dysplasia, carcinoma in situ and invasive squamous cell carcinoma and that aneuploid stem cells may be generated from 2C and 4C stem cell lines in the progression of stromal invasion.     

Loss of hMLH1 expression is associated with less aggressive clinicopathological features in sporadic endometrioid endometrial adenocarcinoma

AIM: To assess the incidence and clinicopathological significance of microsatellite instability (MSI) and the protein expression of hMLH1 and hMSH2 in sporadic endometrioid endometrial adenocarcinoma (SEEA). METHODS: A total of 50 patients with pure endometrioid sporadic endometrial adenocarcinoma were enrolled in the study. MSI analysis was done using five polymorphic markers (BAT26, D5S346, BAT25, D17S250, D2S123) and the protein expression of the hMLH1 and hMSH2 genes was determined by immunohistochemical staining. MSI was detected in 24% (12/50) of SEEA cases. RESULTS: There was a significant correlation between MSI status and loss of hMLH1, hMSH2 expression, respectively. No significant association was found between MSI status and clinicopathological parameters, including age, grade, stage, depth of myometrial invasion, lymph-vascular space invasion (LVI), lymph node involvement or peritoneal cytology. However, significant correlations were found between loss of hMLH1 and a lower histological grade and the absence of LVI in patients with SEEA. CONCLUSIONS: According to these results, MSI and a loss of protein expression of hMLH1 and hMSH2 may be associated with the pathogenesis of SEEA. In addition, hMLH1 immunostaining might have a role as a prognostic parameter. Further research using a large number of cases is needed to confirm our observations.     

Two cases of endometrial cancer meeting new clinical criteria for hereditary nonpolyposis colorectal cancer

BACKGROUND: New clinical criteria for hereditary nonpolyposis colorectal cancer (HNPCC) that includes extracolonic cancers were recently proposed. We present 2 endometrial cancer patients who met the new criteria of 161 endometrial cancer patients. CASE REPORTS: Case 1: A 55-year-old female was operated on for synchronous double primary cancers of the endometrium and rectum. She had also undergone an operation for metachronous ascending colon cancer at the age of 44. She had five relatives with a history of colorectal cancer. The rectal cancer tissue revealed no microsatellite instability (MSI). Case 2: A 48-year-old female underwent a radical operation for synchronous double primary cancers of the endometrium and ovaries. She had three relatives with a history of colorectal cancer. The endometrial cancer tissue showed high MSI. CONCLUSIONS: The frequency of endometrial cancer patients meeting the new HNPCC criteria was 1.2% (2/161). These are the first case reports selected from consecutive endometrial cancer patients.     

不同分子学筛查方法在Lynch综合征相关子宫内膜癌患者中的对比分析#br#

Objective?Compare the accuracy of immunohistochemical staining (IHC), microsatellite instability detection (MSI), and the combined detection of the two in the screening of Lynch Syndrome-related endometrial cancer (LS-EC), and analyze the advantages and disadvantages of actual clinical applications, to identify a routine and reasonable screening strategy. This study also reveals the incidence of LS-EC and the mutations of different mismatch repair (MMR) genes in the Chinese population in this region. Methods?IHC MMR protein detection and MSI detection were performed on the pathological tissues of diagnosed EC patients from the Second Hospital of Jilin University from November 2019 to November 2020, and the preoperative venous blood was subjected to next-generation sequencing (NGS) of Lynch syndrome-related mutations. Results?After NGS testing, 8 cases of LS were confirmed, with an incidence rate of 7.90%. 28 cases had lack of expression of MMR protein, 7 cases were diagnosed as LS, 1 case was missed diagnosis; 13 cases had high microsatellite instability (MSI-H), 3 cases were diagnosed as LS, 5 cases were missed; the combined detection of the two tests can screen out all LS patients. Conclusion?IHC combined with MSI screening is highly sensitive but not cost-effective. IHC alone is recommended as the first clinical screening method.     

Flow cytometric DNA analysis of stage I endometrial carcinoma

Flow cytometric DNA analysis was performed on 203 paraffin-embedded archival specimens obtained from patients with surgical stage I endometrial carcinoma. Primary therapy for those patients (1979-1983) had been definitive extirpation with adjuvant therapy determined by histologic grade, histologic subtype, myometrial invasion, and peritoneal cytologic findings. Diploid DNA patterns were identified in 171 (84%) specimens and nondiploid characteristics were observed in the remaining 32 (25 DNA aneuploid, 7 DNA tetraploid). Although DNA nondiploid specimens accounted for only 16% of all stage I patients, they accounted for 50% of all relapses. Regardless of treatment or other pathologic features, progression-free 5-year Kaplan-Meier survival estimates were 92 and 63% for patients with DNA diploid and DNA non-diploid patterns, respectively (P less than 0.001). Overall 5-year progression-free survival for patients with grade 1 or 2 lesions was 90%; stratification by DNA diploid and DNA nondiploid patterns revealed progression-free survivals of 94 and 64%, respectively (P less than 0.001). Peritoneal cytologic study was positive in seven patients; none of the five with a DNA diploid pattern had a relapse and both with the DNA nondiploid pattern had relapses. These studies suggest that DNA ploidy status may be an objective prognostic determinant for patients with stage I endometrial carcinoma.     

Absence of PTEN repeat tract mutation in endometrial cancers with microsatellite instability

OBJECTIVE: PTEN, a tumor suppressor gene shown to be frequently mutated in endometrial cancers, has been suggested to be a target of microsatellite instability (MSI)-driven mutagenesis. We set out to investigate the relationship between MSI and PTEN mutation in a large series of primary endometrial carcinomas. METHODS: Thirty-nine MSI-positive endometrial cancers were evaluated by single-strand conformational variant analysis and direct sequencing to screen all nine PTEN exons for mutation. RESULTS: Fifteen specimens (38%) demonstrated 16 PTEN mutations. We observed only one alteration in the poly-adenine repeat of exon 8 that is suggested to be a target for mutation in endometrial cancers with MSI. Seven of 16 (44%) mutations in our series were deletions of >/=3 bp, a class of mutation not usually associated with tumors with defective DNA mismatch repair. To determine the significance of this high frequency of deletion, 26 additional endometrial cancers without MSI were matched with the 39 MSI-positive cancers for the prognostic factors of tumor histology, stage, grade, and patient race. The MSI-positive tumors had a significantly higher frequency of deletions involving >/=3 bp when compared with the MSI-negative group (5/11 versus 0/10, P = 0.035). CONCLUSIONS: Repeat tract mutation in PTEN is an uncommon event in MSI-positive cancers. Deletion of >/=3 bp in this gene is more common in MSI-positive cancers when compared with tumors without MSI.     

Pathological findings in early-stage endometrial cancer

OBJECTIVE: The aim of this study was to assess the pathological characteristics of early-stage endometrial cancer, with regard to endometrioid versus serous papillary adenocarcinoma. METHODS: Sixty-six cases of early-stage endometrial carcinoma were classified into two groups: group I--36 cases of endometrioid endometrial cancer, staged IA-IB and graded G1-G2; group II--30 cases of Stage I serous papillary endometrial cancer. The pathological characteristics compared between the two groups included features such as tumor location in the uterine cavity, tumor focality, lymphovascular invasion, as well as the status of the uninvolved endometrium, adjacent to the tumor. Patient clinical characteristics were obtained from the medical records. RESULTS: Significantly more patients with endometrioid endometrial cancer were premenopausal (p < 0.0001), obese (p < 0.02), had hypertension (p < 0.00001) and familial cancer (p < 0.0001). On the other hand, significantly more patients with serous papillary cancer had another primary malignancy (p < 0.001). Considering the pathological characteristics, 75% of endometrioid as compared with 6.7% of serous papillary cancer cases were found in the upper uterine segment only (p < 0.0001). Multifocality was observed in 16.7% of endometrioid as compared with 100% of serous papillary cancer cases (p < 0.0001). Lymphovascular space invasion was absent in all cases of endometrioid cancer, while present in 90% of serous papillary cancer cases (p < 0.0001). Seventy-five percent of endometrioid and 100% of serous papillary cancer cases were associated with an atrophic endometrium. CONCLUSION: The clinical and pathological features of early-stage endometrial cancer differ according to the histological type of the cancer. The majority of endometrioid cancers are probably associated with an atrophic or normally cycling endometrium, and not with endometrial hyperplasia.     

Prognostic significance of DNA ploidy in stage I endometrial cancer

Objective.

To improve the outcome for patients with endometrial cancer, a more accurate prognostic assessment is needed. The current study was undertaken to determine the role of flow cytometric DNA ploidy as an independent prognostic factor in patients with stage I endometrial cancer and to verify if ploidy is able to identify high-risk cases among the apparent ‘low-risk’ patients, defined as stage (IA), grade (1 or 2), and histologic type (endometrioid).

Methods.

This was a retrospective study. DNA ploidy was evaluated from tumor samples in 217 patients with stage I endometrial cancer who underwent definitive surgery as the first treatment between 2003 and 2009. Ploidy and other classic parameters were analyzed in relation to the length of recurrence-free survival.

Results.

Among the 217 evaluated patients, 184 (84.8%) had diploid tumors and 33 (15.2%) had aneuploid tumors. There were 12 recurrences during the median follow-up intervals of 42.7 months. Stage, grade, histologic type, lymphovascular space invasion (LVSI), and ploidy were significantly correlated with recurrence-free interval by univariate Cox analysis. Based on multivariate Cox analysis, ploidy was an independent prognostic factor, with a hazard ratio of 4.5 (95% confidence interval [CI], 1.3-15.3; P = 0.017) adjusted for stage, grade, histologic type, and LVSI. In low-risk patients (n = 156), the recurrence rate was 2.1% for diploid tumors and 12.5% for aneuploid tumors (P = 0.038).

Conclusions.

DNA aneuploidy is an independent prognostic factor in patients with endometrial cancer and can identify high-risk patients among those considered ‘low-risk’ with stage I endometrial cancer.     

DNA aneuploidy is associated with increased mortality for stage I endometrial cancer

OBJECTIVE: The current study was undertaken to determine if DNA ploidy is a useful prognostic variable for predicting recurrence in stage I endometrial cancer. For cancer of the endometrium, survival following recurrence may depend on a number of factors, including the pattern of recurrence and the response to second line treatment. Previous studies have demonstrated a worse survival for patients with DNA aneuploid tumors. It remains unclear, however, whether this is necessarily due to a higher risk of recurrence. This study was undertaken to assess DNA ploidy and risk of recurrence in patients with stage I endometrial cancer. METHODS: This is a retrospective study of surgically treated patients with stages IB and IC endometrial cancer treated from 1992 to 2000. All patients underwent definitive surgery, including staging lymphadenectomy. None of the patients received postoperative treatment. DNA ploidy was determined using flow cytometry and image analysis. Grade, lymph-vascular space invasion, stage (stage IB versus IC), and DNA ploidy were analyzed with regard to recurrence and survival. RESULTS: There were 100 patients with stages IB and IC endometrial cancer in this analysis. There were 17 recurrences (17%) and 10 patients that died of cancer (10%). Grade 3 and the presence of lymph-vascular space invasion were associated with increased risk of recurrence; DNA aneuploidy and stage were not. Grade, lymph-vascular space invasion, and DNA ploidy were associated with survival. These findings indicate that DNA aneuploidy does not increase the risk of disease recurrence but is associated with overall survival. CONCLUSION: Although the recurrence risk is not higher for patients with surgical stage I endometrial cancer and aneuploid tumors, overall mortality remains higher.     

Microsatellite instability,MLH1 promoter methylation,and loss of mismatch repair in endometrial cancer and concomitant atypical hyperplasia

OBJECTIVE: MLH1 methylation is associated with the microsatellite instability (MSI) phenotype in endometrial cancer and atypical endometrial hyperplasia, a premalignant precursor to carcinoma. The observation that methylation is also seen in atypical endometrial hyperplasia without MSI suggests that methylation is an early event in endometrial tumorigenesis. Our objective was to determine if methylation is always present in MSI-positive atypical hyperplasia concomitant with MSI-positive, methylation-positive carcinoma. METHODS: We used laser capture microdissection to study MLH1 methylation and MSI in a large series of endometrial cancer cases that had previously been shown to have methylation and the MSI-high (MSI-H) phenotype. We resampled areas of carcinoma from 27 patients along with 51 foci of concomitant atypical endometrial hyperplasia. RESULTS: Consistent with previous reports, we saw MLH1 methylation in areas of atypical endometrial hyperplasia that did not show MSI. In addition, we noted that 18% of the MSI-H atypical endometrial hyperplasia DNAs lacked methylation of critical cytosines in the MLH1 promoter. Immunohistochemistry studies showed that these MSI-H unmethylated foci of atypical endometrial hyperplasia failed to express MLH1, as did regions of simple hyperplasia. CONCLUSION: Methylation of the MLH1 promoter is an early event in endometrial tumorigenesis. Given that not all MSI-positive tissues had methylation at cytosines -229 and -231, it appears that methylation may not be required for MLH1 silencing and loss of mismatch repair.     

Chemosensitivity and DNA content as prognostic factors in ovarian cancer: preliminary results

Fourteen samples of epithelial ovarian cancer were studied; chemosensitivity in vitro was examined by clonogenic assay, and DNA content was measured by static cytometry. Seven cases were aneuploid. The average survival of the diploid cases was almost twice that of the aneuploid cases (21 vs 11 months). Five of the 7 diploid, and 4 of the aneuploid cases were chemosensitive in vitro; 4 diploid and 3 aneuploid tumors were sensitive in vivo. The vitro/vivo correlation was positive for 6 out of 7 aneuploid tumors, and for 4 out of 7 diploid cases. Patients sensitive in vitro to at least one of the two drugs tested responded more often to chemotherapy, whatever drugs were used in vitro. The patients whose 5 C exceeding rate was 10% survived 12 months at most, with an average survival of 8 months; patients with a low aneuploidy index survived an average of 20 months. The behavior of the diploid tumors in vitro was more varied. Ploidy would seem to be an additional factor to other parameters of prognosis, while no correlation is evident between chemosensitivity in vitro, D.I., and 5 C exceeding rate.     

High concordance of molecular tumor alterations between pre-operative curettage and hysterectomy specimens in patients with endometrial carcinoma

Objective

Molecular alterations in endometrial cancer have been shown to be prognostically significant but have not yet been implemented in the current clinical risk assessment. Few studies have investigated the reliability of molecular alterations in pre-operative specimens. Therefore, the objective was to determine whether molecular analysis of pre-operative endometrial cancer samples accurately reflects those alterations in the subsequent hysterectomy specimens.

Methods

Paired pre-operative and hysterectomy specimens of 48 patients diagnosed with endometrial carcinoma, 42 endometrioid (EEC) and 6 non-endometrioid (NEEC) carcinomas, were analyzed for immunohistochemical expression of p53, PTEN and β-catenin. Tumor DNA was isolated and analyzed for microsatellite instability (MSI), TP53 mutations and somatic hot spot mutations in 13 genes.

Results

In EEC patients, loss of PTEN, nuclear β-catenin and p53-mutant expression was found in 43%, 7% and 12%, respectively. No nuclear β-catenin was found in 5 of 6 NEEC patients, all serous cancers, whereas a p53-mutant expression was present in all serous cases. MSI was found in 19.5%, all EEC. Concordance for PTEN, β-catenin, p53 expression and MSI status was found in 79%, 92%, 79% and 93.5%, respectively. We detected 65 hot spot mutations in 39/48 (81%) tumors. Overall concordance of the GynCarta multigene analysis was 99.8%.

Conclusions

The results confirm the reliability of immunohistochemical and DNA-based techniques in the evaluation of molecular alterations in pre-operative endometrial specimens and high concordance rates with the definitive hysterectomy specimens. The resulting molecular signature provides initial pre-operative diagnostic information on the status of oncogenic pathways, which may contribute to individualized treatment strategies.