Node-negative Cases Research Articles

PWE-004 Detection of Venous Invasion to Stage Colorectal Cancer

IntroductionElastica staining of tumour sections increases the sensitivity of detection of venous invasion in colorectal cancer. We compared the prognostic value of elastica detected venous invasion with that of other...

Clinical study of sentinel lymph node detection guided radical abdominal trachelectomy

To evaluate the clinical value of sentinel lymph nodes (SLN) in predicting pelvic lymph node status for early cervical squamous cell carcinoma, and approach the clinical significance of SLN detection for guiding radical abdominal trachelectomy (RAT).Outcomes of follow up and fertility were also observed. A total of 31 patients with stageIa2-Ib1 squamous cell carcinoma planned to be given RAT and pelvic lymphadenectomy were enrolled. (99m)Tc-labeled phytate was injected before surgery.Intraoperatively, SLN were identified, excised, and submitted to fast frozen section.Systematic bilateral pelvic lymphadenectomy was performed, and then RAT was performed in patients with negative SLN. All nodes were sent for routine pathological examination and immunostained with anti-cytokeratin antibody to detect micrometastases. SLN were detected in all patients (100%,31/31). A total of 109 SLN were identified with a mean number of 3.5 per patient.Of these, SLN of 2 patients were positive on frozen sections and proved to be metastasis by final pathologic examination and quitted the RAT. No missed micrometastasis was found using immunohistochemical staining in SLN and other lymph nodes using histologically node-negative cases. No false negative cases was found and the negative value was 100% (31/31). The sensitivity, accuracy, and false negative rates were 100%, 100%, and 0, respectively. Perioperative complications occurred in 5 patients including 2 cases of bladder injury and 3 cases of uterine artery injury.No relapses occurred during follow-up.Five of 19 patients with procreative desire conceived pregnancies (4 spontaneous abortion and 1 premature birth) after surgery. The identification of SLN using (99m)Tc-labeled phytate could predict the pelvic lymph node status in early stage cervical cancer. Under the guidance of SLN detection, RAT is a feasible operative modality with well prognosis and low complications for young patients who desire to preserve reproductive function.

Thoracoscopic esophagectomy with extended lymph node dissection in the left lateral position: technical feasibility and oncologic outcomes

The aim of this study was to estimate the technical and oncologic feasibility of video-assisted thoracoscopic radical esophagectomy (VATS) in the left lateral position. From January 2003 to December 2011, 132 patients with esophageal cancer underwent VATS. The mean duration of the thoracic procedure and the entire procedure was 294 ± 88 and 623 ± 123 minutes, respectively. Mean blood loss during the thoracic procedure and the entire procedure was 313 ± 577 and 657 ± 719 g, respectively. The mean number of dissected thoracic lymph nodes was 32.6 ± 12.9. There were four in-hospital deaths (3.0%); two patients (1.5%) died of acute respiratory distress syndrome and two patients (1.5%) died of tumor progression. Postoperative unilateral or bilateral recurrent laryngeal nerve (RLN) palsy, or pneumonia was found in 33 (25.0%), 21 (15.9%), and 27(20.5%) patients, respectively. The patients were divided into the first 66 patients who underwent VATS (Group 1) and the subsequent 66 patients (Group 2). The numbers of cases who underwent neoadjuvant or induction chemotherapy for T4 tumor and intrathoracic anastomosis were higher in Group 2 than in Group 1. The duration of the procedure, amount of blood loss, and the number of dissected thoracic lymph nodes were not different between the two groups. The total number of dissected lymph nodes was higher in Group 2 than in Group 1 (72.6 ± 27.8 vs. 62.6 ± 21.6, P = 0.023). The rate of bilateral RLN palsy was less in Group 2 than in Group 1 (7.6% vs. 24.2%, P = 0.042). The mean follow-up period was 38.7 months. Primary recurrence consisted of hematogenous, lymphatic, peritoneal dissemination, pleural dissemination, and locoregional in 15 (11.3%), 20 (15.1%), 3 (2.3%), 4 (3.0%), and 5 patients (3.8%), respectively. The rate of regional lymph node recurrence within the dissection field was only 4.5%. The prognosis of patients with lymph node metastasis was significantly poorer than that of patients without lymph node metastasis. However, the prognosis of the 11 cases that had metastasis only around RLNs was similar to that of node-negative cases. Thirteen patients with pathological remnant tumor (R1 or R2) did not survive longer than 5 years at present. The overall 5-year survival rate of stage I, II, and III disease after curative VATS was 82.2%, 77.0%, and 52.3%, respectively. Expansion of VATS criteria for patients after induction chemotherapy for T4 tumor or thoracoscopic anastomosis did not adversely affect the surgical results by experience. Although the VATS procedure is accompanied by a certain degree of morbidity including RLN palsy and pulmonary complications, VATS has an excellent locoregional control effect. In addition, the favorable survival after VATS shows that the procedure is oncologically feasible.

Comparison of microscopic (pT3a) and gross extravesical extension (pT3b) in pathological staging of bladder cancer: analysis of patient outcomes

To determine the prognostic value of pT3 bladder urothelial carcinoma substaging in patients without lymphatic involvement. Pathologic and clinical data were reviewed on patients who underwent radical cystectomy for urothelial carcinoma between 1991 and 2010. Of the 460 reviewed patients, 74 patients were diagnosed with pathologic T3No urothelial carcinoma of the bladder. The impact of pathologic substaging (pT3a vs. pT3b) was examined to determine the effect on overall and disease-specific survival. Five years disease-specific and overall survival rates were 46.9 % and 39.6 % for patients with pT3aNo tumor, whereas these ratios were 34.4 and 30.3 %, respectively, for patients with pT3bNo tumor (p > 0.05). Mean disease-specific survival time was 43.94 ± 6.50 months for pT3aNo, while it was 39.01 ± 7.19 months for pT3bNo (p = 0.539). In multivariate cox regression analysis, age (p = 0.459), gender (p = 0.710), urinary diversion type (p = 0.088), and pT3 substaging (p = 0.554) were not noticed as an independent predictive factor for survival. Macroscopic extravesical extension (pT3b) is not associated with a worse outcome than pT3a disease in lymph node-negative cases of bladder urothelial carcinoma.

Prognostic significance of histologically detected lymph node micrometastases of sizes between 0.2 and 2 mm in colorectal cancer

Minimal lymph node involvement is a potential prognostic factor in colorectal cancer. The International Union Against Cancer defined tumour deposits between 0.2 and 2 mm as micrometastases and clusters and single-cell infiltrations below this cutoff as isolated tumour cells. Nevertheless, only a minority of studies discriminated metastatic involvements according to this definition. In order to investigate the prognostic significance of micrometastases (0.2-2 mm), we performed a retrospective study enrolling 44 routinely diagnosed micrometastatic cases within 15 years which represent about 1% of our cases. These cases have been re-evaluated. Seven of the micrometastatic cases turned out to be macrometastases after step sectioning. Complete follow-up was available in 33 remaining cases. Collections of node-negative and macrometastatic cases served as control groups. The Kaplan-Meier curves of macro- and micrometastatic cases showed a similar adverse course (p = 0.830) especially during the first 40 months. The 5-year-survival rates were 51, 60 and 64 months for macro-, micrometastatic and node-negative cases, respectively. The difference in overall survival, however, reached only a statistical trend and was not significant (p = 0.137). After re-evaluation with step sections and cytokeratin immunohistochemistry out of an initial 91 node-negative cases, 11 (12%) cases were identified with isolated tumour cells and one (1%) case with a micrometastasis. Our data show that micro- and macrometastatic colorectal cancers show very similar survival rates. Therefore, efforts to improve the detection of lymph node micrometastases seem to be justified.

Validation of the 7th TNM classification for non-small cell lung cancer: A retrospective analysis on prognostic implications for operated node-negative cases

Background. The 7th TNM staging system for non-small cell lung cancer (NSCLC) developed by the International Association for the study of Lung Cancer (IASLC) has been applied in Sweden since the beginning of the year 2010. The aim of this retrospective study was to evaluate the prognostic role of the 7th TNM staging system in a surgical Swedish patient cohort with node-negative NSCLC. Material and methods. We collected data from stage I patients (pT1-2 pN0, 6th TNM system) who underwent surgery for NSCLC at Karolinska University Hospital from 1987 to 2002. Tumors were restaged according to the 7th TNM version. Cox multivariate survival analysis was implemented in order to determine the prognostic impact of pathological stage when classified according to either the 6th or the 7th TNM systems. Results. The patient population consisted of 452 subjects. Tumor size was ≤ 3 cm in 51% of cases. The predominant histology was adenocarcinoma (53%) and lobectomy was the most common surgical procedure (82% of patients). The five-year survival rate in patients with stage IA vs. IB (6th TNM) was 62% vs. 51%, respectively (log-rank p = 0.036). Corresponding figures for the 7th TNM system were 70% in stage IA-T1a, 51% in stage IA-T1b, 54% in stage IB, 51% in stage IIA and 35% in stage IIB (log-rank p = 0.002). On multivariate analysis, adjusted by age, gender, histology, kind of surgery, grade of differentiation and smoking status, pathological stage was an independent prognostic factor if classified according to the 7th TNM version (p = 0.001), but not if scored according to the 6th TNM edition (p = 0.090). Conclusion. The 7th TNM classification system is a more accurate predictor of prognosis in stage I operated patients than the old classification. The new system should be implemented even on retrospective cohorts especially when investigating the prognostic implication of the expression of molecular biomarkers.

Value of Sentinel Lymph Node Biopsy in Breast Cancer Surgery with Simple Pathology Facilities -An Iranian Local Experience with a Review of Potential Causes of False Negative Results

Sentinel lymph node biopsy (SLNB) is a precise procedure for lymphatic staging in early breast cancer. In a valid SLNB procedure, axillary lymph node dissection (ALND) can be omitted in node- negative cases without compromising patient safety. In this study, detection rate, accuracy and false negative rate of SLNB for breast cancer was evaluated in a setting with simple modified conventional pathology facilities without any serial sectioning or immunohistochemistry. Patients with confirmed breast cancer were enrolled in the study. SLNB and ALND were performed in all cases. Lymph node metastasis was evaluated in SLN and in nodes removed by ALND to determine the false negative rate. Pathologic assessment was carried out only by modified conventional technique with only 3 sections. Detection rate was determined either by lymphoscintigraphy or during surgery. 78 patients with 79 breast units were evaluated. SLN was detected in 75 of 79 cases (95%) in lymphoscintigraphy and 76 of 79 cases (96%) during surgery. SLN metastases was detected in 30 of 75 (40%) cases either in SLNB and ALND groups. Accuracy of SLNB method for detecting LN metastases was 92%. False negative rate was 3 of 30 of positive cases: 10%. In 7 of 10 cases with axillary lymphadenopathy, LN metastastates was detected. SLNB is recommended for patients with various tumor sizes without palpable lymph nodes. In modified conventional pathologic examination of SLNs, at least macrometastases and some micrometastases could be detected similar to ALND. Consequently, ALND could be omitted in node-negative cases with removal of all palpable LNs. We conclude that SLNB, as one of the most important developments in breast cancer surgery, could be expanded even in areas without sophisticated pathology facilities.

Detection of promoter methylation of tumor suppressor genes in serum DNA of breast cancer cases and benign breast disease controls

Tumors are capable of shedding DNA into the blood stream. This shed DNA may be recovered from serum or plasma. The objective of this study was to evaluate whether pyrosequencing promoter DNA in a panel of 12 breast cancer-related genes (APC, BRCA1, CCND2, CDH1, ESR1, GSTP1, HIN1, P16, RARβ, RASSF1, SFRP1 and TWIST) to measure the degree of methylation would lead to a useful serum-based marker of breast cancer. Serum was obtained from women who were about to undergo a breast biopsy or mastectomy at three hospitals from 1977 to 1987 in Grand Rapids, MI USA. We compared the methylation status of 12 genes in serum DNA obtained from three groups of postmenopausal women (mean age at blood collection: 63.0 y; SD 9.9; range 35–91): breast cancer cases with lymph node-positive disease (n = 241); breast cancer cases with lymph node-negative disease (n = 63); and benign breast disease control subjects (n = 234). Overall, median levels of promoter methylation were low, typically below 5%, for all genes in all study groups. For all genes, median levels of methylation were higher (by 3.3 to 47.6%) in lymph node-positive breast cancer cases than in the controls. Comparing mean methylation level between lymph-node positive cases and controls, the most statistically significant findings, after adjustment of the false-positive rate (q-value), were for TWIST (p = 0.04), SFRP1 (p = 0.16), ESR1 (p = 0.17), P16 (p = 0.19) and APC (p = 0.19). For two of these four genes (TWIST, P16), the median methylation level was also highest in lymph-node positive cases, intermediate in lymph node-negative cases and lowest in the controls. The percent of study subjects with mean methylation scores ≥ 5% was higher among lymph node-positive cases than controls for ten genes, and significantly higher for HIN1 and TWIST (22.0 vs. 12.2%, p = 0.04 and 37.9 vs. 24.5%, p = 0.004, respectively). Despite relatively consistent variation in methylation patterns among groups, these modest differences did not provide sufficient ability to distinguish between cases and controls in a clinical setting.

Diagnosis and treatment of male breast cancer

SUMMARY Male breast cancer (MBC) accounts for <1% of mammary neoplasia and because of its rarity, diagnosis and treatment is based on breast cancer in women. The PubMed database was searched for all English language articles from 1993 to May 2012. Search terms included: male breast cancer, diagnosis, treatment and management. Although rare, MBC incidence is rising. Risk factors include hormonal imbalance, Klinefelter’s syndrome, exposure to radiation and BRCA2 mutations. Clinical evaluation combined with fine-needle aspiration/core biopsy usually suffices for diagnosis, but mammography can be helpful. Modified radical mastectomy was the standard treatment; total mastectomy and sentinel node biopsy is now the treatment of choice in node-negative cases. Tamoxifen is the mainstay of adjuvant hormonal therapy, but a role for aromatase inhibitors is emerging. Given the low incidence of MBC, worldwide studies are needed to improve management and prognosis.

Predictors of Lymph Node Count in Colorectal Cancer Resections

To identify factors that influence the total and negative lymph node counts in colorectal cancer resection specimens independent of pathologists and surgeons. We used multivariate negative binomial regression. Covariates included age, sex, body mass index, family history of colorectal carcinoma, year of diagnosis, hospital setting, tumor location, resected colorectal length (specimen length), tumor size, circumferential growth, TNM stage, lymphocytic reactions and other pathological features, and tumor molecular features (microsatellite instability, CpG island methylator phenotype, long interspersed nucleotide element 1 [LINE-1] methylation, and BRAF, KRAS, and PIK3CA mutations). Two US nationwide prospective cohort studies. Patients with rectal and colon cancer (N=918). The negative and total node counts (continuous). Specimen length, tumor size, ascending colon location, T3N0M0 stage, and year of diagnosis were positively associated with the negative node count (all P.002). Mutation of KRAS might also be positively associated with the negative node count (P=.03; borderline significance considering multiple hypothesis testing). Among node-negative (stages I and II) cases, specimen length, tumor size, and ascending colon location remained significantly associated with the node count (all P.002), and PIK3CA and KRAS mutations might also be positively associated (P=.03 and P=.049, respectively, with borderline significance). This molecular pathological epidemiology study shows that specimen length, tumor size, tumor location, TNM stage, and year of diagnosis are operator-independent predictors of the lymph node count. These crucial variables should be examined in any future evaluation of the adequacy of lymph node harvest and nodal staging when devising individualized treatment plans for patients with colorectal cancer.

Impact of the new FIGO 2009 staging classification for vulvar cancer on prognosis and stage distribution

ObjectiveIn 2009, FIGO modified staging of vulvar cancer — the performance of the new classification relative to the prior system has not been assessed. We sought to investigate the impact of the 2009 FIGO vulvar cancer staging system on stage distribution and prognostic ability of the 2009 sub-stage classifications in a large cohort of uniformly staged cases with long-term followup. MethodsPatients undergoing surgery for vulvar cancer were identified from 2 institutions (Mayo Clinic and Medical University, Gdansk, Poland) using a similar surgical approach. Inclusion criteria required primary surgery for invasive vulvar cancer for cases with >1mm invasion with complete inguinal/femoral lymphadenectomy. The technique of inguinofemoral node dissection used in both institutions was designed to remove both superficial and deep inguinofemoral nodes. A retrospective review was performed and all cases were assigned stage using the 1988 and 2009 FIGO systems after reviewing pathology slides. Cause-specific survival (CSS, death due to cancer) was estimated using the Kaplan–Meier method and compared using the Cox proportional hazards model t for the first 10years after surgery. ResultA total of 468 patients met inclusion criteria. Thirty-one percent (n=155) were down-staged, and 1 case up-staged using 2009 staging. The new system fails to effectively separate 10-yr CSS for stage I and II cases (p=0.52), while FIGO 1988 failed to separate stages II and III (p=0.41). We observed a difference in survival for stage I and II cases based on tumor diameter. For smaller stage II lesion (≤4cm vs. >4cm) we observed no difference in survival compared to all stage IB cases (p=0.25) Considering node positive disease, patients with 2009 FIGO stages ΙΙΙA, ΙΙΙB, and ΙΙΙC were not significantly different in terms of CSS (p=0.17). However, CSS approached significance between patients with extracapsular vs. intracapsular disease (p=0.072). For stages IIIA and IIIB (excluding extracapsular spread, IIIC), we observed that the number of positive nodes and diameter of lymph node metastasis were not significantly associated with CSS. When comparing bilateral nodal involvement vs. unilateral cases with at least 2 involved nodes, we found no statistical difference in CSS (p=0.30). ConclusionThis is the largest cohort study to evaluate the effect and prognostic performance of the new FIGO vulvar cancer staging system. The new staging does not stratify survival between stages I and II and reduces CSS in stage I cases. Our results suggest that lesion size in node negative cases is an important prognostic variable that could be addressed in future staging classifications. Among the node positive cases, the current classification results in slight differences in CSS, primarily between intra- and extra-capsular disease and not according to the number of positive nodes and lymph node metastasis diameter. Finally we observe that bilateral nodal disease does not appear to impact CSS, justifying it being omitted from the 2009 staging system and that separating node positive (2009 stage III) from node negative (2009 stage II) cases is justified.

Diagnosis of axillar lymph node status and sentinel node biopy (SNB) by indocyanin green (ICG) fluorescence imaging in early breast cancer patients.

e11503 Background: In breast cancer patients, diagnosis of Axillar lymph node (Ax LN) status is practical use in deciding treatment strategy. But, diagnostic methods are different by different institution. The purpose of this study was to review diagnostic methods of Ax LN status. Methods: From December 2006, 483 cases were underwent breast cancer operation and within 356 (74%) clinical node negative primary breast cancer cases underwent Sentinel Node biopsy (SNB) by indocyanine green (ICG) fluorescence imaging. Dye was injected sub dermal in the areola. After one minute massage, subcutaneous lymphatic channel draining from the areola to the axilla was observed by Image processing and measuring system (PDE C9830, Hamamatsu Photonics Co). Skin incision was done at the point which fluorescence imaging disappeared (ASCO2011, Electronic Publication only). From September 2010, Aspiration Biopsy Cytology (ABC) of Ax LN (LN-ABC), Ultrasonography (US) and CT were done as routine workup in breast cancer patients. Node positive cases were received pre-operative chemotherapy or Ax LN dissection and node negative cases were underwent SNB by ICG fluorescence imaging during operation. Results: From December 2006, SNB were done 356 clinical node negative primary breast cancer cases. SN was detected 342 cases, detective rate was 96 %. This rate was as good as SNB done by radioactive colloid (Phytate radiolabelled by Tc98) with dye (detective rate was 95%, ASCO2011). The lymph node metastasis was detected 48 cases (14%). From Sep 2010 to Oct 2011, LN-ABC was done 49 cases of the 142 primary breast cancer patients. Fourteen cases were LN-ABC positive and 35cases were LN-ABC negative. Positive SNB were detected 3 of the 35 cases (2 cases had micro metastasis and one case had macro metastasis). Within three SNB positive cases, pre-operative US and CT had no positive finding. Positive LN-ABC cases (14 cases) were received pre-operative therapy or Ax LN dissection. Conclusions: These results suggest that LN-ABC was useful procedure detecting positive Ax LN and SNB by ICG fluorescence imaging was useful and easy tool in operative breast cancer patients.

Factors predictive of papillary thyroid micro-carcinoma with bilateral involvement and central lymph node metastasis: a retrospective study

BackgroundThe optimal resection extent for papillary thyroid microcarcinoma (PTMC) remains controversial. The objective of the study was to investigate risk factors of bilateral PTMC and central lymph node metastasis (CLNM) to guide surgical strategies for PTMC patients.MethodsWe retrospectively reviewed 211 PTMC patients who underwent total thyroidectomy (TT) and 122 clinical lymph node-negative (cN0) cases that underwent prophylactic central lymph node dissection (CLND) between 2010 and 2011. The frequency, pattern, and predictive factors for bilateral PTMC and CLNM in these patients were studied using univariate and multivariate analysis with respect to the following variables: age, gender, extrathyroidal extension (ETE), T stage, with Hashimoto thyroiditis (HT), tumor size and multifocality based on final pathology, and preoperative evaluation using ultrasonography (US).ResultsFifty-four of 211 (25.6%) patients had bilateral PTMC. In multivariate analysis, multifocality (P < 0.001, OR = 23.900) and tumor size ≥7 mm (P = 0.014, OR = 2.398) based on US were independent predictive factors for bilateral PTMC which was also independently associated with multifocality (P < 0.001, OR = 29.657) and tumor size ≥7 mm (P = 0.005, OR = 2.863) based on final pathology. Among 122 cN0 patients who underwent prophylactic CLND, we found 49.2% of patients had CLNM. CLNM was independently associated with men, age <50 years and tumor size ≥7 mm based on final pathology or preoperative US.ConclusionsTT should be considered for PTMC patients who are found multifocality and tumor size ≥7 mm based on preoperative US. CLND need be considered in cN0 patients who are men, aged <50 years or tumor size ≥7 mm based on preoperative US.

Abstract 4201: Analysis of gene expression profile and signaling networks potentially involved in lymph node metastasis in esophageal squamous cell carcinoma

Abstract Esophageal squamous cell carcinoma (ESCC) is the 8th most common cancer worldwide and the 6th tumor in frequency in Brazil. Most of cases are diagnosed at an advanced stage and despite the improvement of various treatment modalities the overall survival rates still remain low. The aim of this study was to analyze the gene expression profile to identify the genes related to lymph node metastasis in ESCC samples. Genome-wide expression profiling was carried out in total RNA extracted from 17 frozen tissue samples using the Agilent Whole Human Genome Microarray 44K following the manufacturer's protocol. Data were extracted and flagged with Feature Extraction and processed using TMEV Software. A supervised hierarchical clustering was performed using the Pearson distance metric. Paired t-test analysis with 1000 permutations was performed to generate a list of differentially expressed genes from the comparison between the different groups. Only genes showing a false discovery rate (Q value) &amp;lt;5% were accepted as differentially expressed. The Ingenuity Pathway Analysis (IPA) Software was used to assess the involvement of significantly differentially expressed genes in known pathways and networks. Hierarchical clustering showed two distinct groups according to lymph node metastasis: Group 1 comprised 6 node-negative cases and Group 2 with 11 node-positive cases. The gene expression profiling revealed 338 genes differentially expressed between the two groups, of which 321 showed lower expression and 17 showed a higher expression in cases of Group 2 when compared to those of Group 1. According to IPA analysis five significant networks could be defined comprising the genes differentially expressed between the two groups. The first network with a score of 43 comprised 21 of the differentially expressed genes and was associated to functions as cell-to-cell signaling and interaction, lipid metabolism and molecular transport. The second network (score 22, 13 genes) was related to nucleic acid metabolism, small molecules biochemistry and energy production. The third network (score 21, 14 genes) was associated to cellular signaling, molecular transport and small molecular biochemistry. The network 4 (score of 21, 12 genes) was associated to cancer, DNA replication, recombination and repair and gene expression. The fifth network (score 21, 12 genes) was related to cellular movement, cardiovascular system development and function and neurological disease. In this study, it was detected genes differentially expressed between cases with and without lymph node metastasis involved in important signaling networks that could have an impact on tumor development or progression in ESCC. The identified genes might be useful as a first step to identify molecular markers to predict lymph node metastasis for improved diagnostic and therapeutic modalities in this tumor. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4201. doi:1538-7445.AM2012-4201

P3-06-06: Comparison of Gene Expression Profiles of Lymph Node Positive and Lymph Node Negative ER Positive Breast Tumors in Pre- and Postmenopausal Women.

Abstract Background Breast cancer is the most common female cancer in US and is the second leading cause of cancer related death in women. Metastases are the primary cause of cancer morbidity and mortality. Axillary lymph node (LN) status has long been used as a prognostic factor for breast cancer. The molecular mechanisms that control LN metastasis remains poorly understood. To better understand the various genes and regulatory pathways that drive breast cancer LN metastasis, we compared the gene expression profiles between breast tumors that have metastasized to the LNs and those which have not in pre- and postmenopausal women. Material and Methods: Tumor cells were isolated from the primary tumors (ER+) of postmenopausal node positive (PMNP; N=20), postmenopausal node negative (PMNN; N=19), premenopausal node positive (PRNP; N=18) and premenopausal node negative (PRNN; N=16) women using laser capture microdissection. RNA was isolated using the RNAqueous®-micro kit (Ambion, Austin, TX). Total RNA was converted to Biotin-labelled aRNA using two rounds of amplification with MessageAmp II aRNA amplification kit (Applied Biosystems, Foster City, CA). The aRNA concentration was determined by Nanodrop 1000 and the quality was assessed with a Bioanalyzer. The aRNA was fragmented and hybridized to Human Genome U133 Plus 2.0 GeneChip (Affymetrix, Santa Clara, CA). Microarray raw data were analyzed using a variety of R programming packages for probe density processing, background correction, normalization, quality control/quality assessment, and calculation of gene expression value, etc. To identify differentially expressed genes, Wilcoxon rank sum test with FDR (false discovery rate) control was performed for pair-wise comparison between different groups. Functional analyses were performed on the identified statistically significant differentially expressed genes to search for the functional categories and pathways in which they are involved and further understand their potential roles in breast cancer metastatic process. Results: Multivariate data mining (hierarchical clustering analysis and principal component analysis, etc) revealed that in postmenopausal women, the node positive and node negative women are well separated while this was not the case in premenopausal women. Further analysis of the PMNN and PMNP groups to identify differentially expressed genes (with at least a 1.5 fold difference) at FDR =0.1 showed that 232 genes were upregulated and 470 genes were downregulated in PMNP vs PMNN groups. Gene function analysis revealed that genes down regulated in the PMNP group compared to PMNN are related to extracellular matrix, cell adhesion, EGF-like pathway, cytoskeleton etc, while the over-expressed genes are related to cell cycle and cell division, chromosome condensation, etc. Discussion: The ability to differentiate lymph node positive cases from lymph node negative cases in ER+ breast cancer based on transcriptional profiling may have an impact on the clinical management of ER+ breast cancer cases. Having transcriptional profiles that identify ER+ tumors likely to have poor outcomes would suggest more aggressive treatment for such patients. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-06-06.

P5-01-13: High Levels of Nuclear Heat Shock Factor 1 (HSF1) Are Associated with Poor Prognosis in Breast Cancer: Results from the Nurses' Health Study.

Abstract Purpose: Heat shock factor 1 (HSF1) is the master transcriptional regulator of the cellular response to heat and a wide variety of other stressors. We previously reported that HSF1 promotes the survival and proliferation of malignant cells. At this time, however, the clinical and prognostic significance of HSF1 in cancer is unknown. Patients and methods: Breast cancer samples from 1,841 participants in the Nurses’ Health Study (NHS) were scored for levels of nuclear HSF1. Associations of HSF1 status with clinical parameters and survival outcomes were investigated by Kaplan-Meier analysis and Cox proportional hazard models. The associations were further delineated by Kaplan-Meier analysis using publicly available mRNA expression data. Results: Nuclear HSF1 levels were elevated in ∼80% of in situ and invasive breast carcinomas. In invasive carcinomas, HSF1 expression was associated with high histologic grade, larger tumor size, and nodal involvement at diagnosis (P&amp;lt;0.0001). Overall, in multivariate analysis, high-HSF1 levels were associated with increased breast cancer-specific mortality (HR, 1.62; 95% CI, 1.21−2.17). This association was seen in the ER-positive population (HR, 2.10; 95% CI, 1.25−2.47), even in early-stage lymph node negative cases (HR, 1.98; 95% CI, 1.17−3.33). In public expression profiling data, high-HSF1 mRNA levels were also associated with an increase in ER-positive breast cancer-specific mortality. Conclusions: Increased HSF1 is associated with reduced survival in breast cancer. The findings indicate that HSF1 should be evaluated prospectively as an independent prognostic indicator in ER-positive breast cancer and that HSF1 may provide a useful therapeutic target. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-01-13.

Occult tumor metastasis and the prognostic value of sentinel lymph nodes in rectal cancer

The aim of the present study to evaluate the application of sentinel lymph node (SLN) pathology in rectal cancer using ex vivo mapping and to investigate the incidence and prognostic value of occult SLN metastasis in routine node-negative specimens. Specimens (n=117) of rectal cancer were examined using a combination of routine pathology and ex vivo SLN mapping. The inspected SLNs were further treated with immunohistochemical staining for occult cancer foci. The log-rank test was used to assess survival. SLNs were examined in 112 of the included specimens with a total number of 212, resulting in an identification rate of 95.7% (112/117). The status of SLNs accurately reflected N stage in 93.8% (105/112) of cases and the sensitivity was 75.6% (31/41) in detecting nodal metastasis. The accuracy of SLN pathology decreased in cancers of more advanced TNM stages (P=0.001). In 74 cases with routine node-negative (N0) disease, SLN micrometastasis (MIC) and isolated tumor cells (ITC) were examined in 9 and 4 cases, while the remaining 61 were regarded as negative (NEG). The log-rank test revealed poorer disease-free and overall survival of the MIC group compared with the NEG group. However, the findings from the ITC group were not significant compared with those from the MIC and NEG groups. In conclusion, ex vivo SLN pathology is practical and accurate in rectal cancer; however, its utility is limited in cases of advanced disease. Immunohistochemically detected SLN micrometastasis in node-negative cases is therefore a predictor of poorer outcome, and may therefore be listed as a marker for adjuvant therapy.

Sentinel node biopsy (SNB) by indocyanin green (ICG) fluorescence imaging in patients with operative breast cancer.

e11507 Background: Sentinel node biopsy (SNB) is standard method of breast cancer operation. There are two methods in detecting SNB using dye only and radioactive colloid with dye in Japan. We evaluated usefulness of SNB by indocyanin green (ICG) fluorescence imaging in operative breast cancer patients, retrospectively. Methods: From December 2006 to December 2010, 360 cases underwent breast cancer operation and within 253 (70%) casesin clinical node-negative cases underwent SNB by ICG fluorescence imaging. Dye was mixed 1ml ICG and 4ml Indigocarmine. Dye was injected subdermal in the areola. After one minute massage, subcutaneous lymphatic channel draining from the areola to the axilla was observed by Image processing and measuring system (PDE C9830, Hamamatsu Photonics Co). Skin incision was done at the point which fluorescence imaging disappeared. Results: SN was detected 238 cases, detective rate was 96%.The lymph node metastasis was detected 37 cases (15%). Author has been belonged at another hospita...

Abstract 85: Detection of promoter methylation of tumor supressor genes in serum DNA of breast cancer patients and controls

Abstract Background: Changes in DNA methylation patterns are a common feature of malignant cells. Methylation is an epigenetic change that affects gene expression. Methylation of a cytosine guanine dinucleotide-rich area (CpG island) of the promoter region of a gene that is normally unmethylated is correlated with silencing of that gene. Certain tumor suppressor genes are hypermethylated in breast tumor tissue and rarely methylated in normal breast tissue. Circulating serum DNA, presumably including DNA shed from the tumor, can be tested for promoter hypermethylation. Breast cancer detection with moderate sensitivity has been achieved in several small studies relying on a few genes and suggests that assaying methylation levels in a larger panel of genes might result in a useful serum marker for breast cancer detection. Methods: Bisulfite-modified serum DNA was evaluated for promoter methylation via pyrosequencing in 12 candidate tumor suppressor genes (GSTP1, SRFP1, RAR-beta, ER1, TWIST, HIN1, APC, ECAD, p16, BRAC1, CyclinD2, RASSF1). Study subjects included approximately 250 node-positive postmenopausal breast cancer cases, 75 node negative postmenopausal breast cancer cases and a comparison group of 250 postmenopausal women with benign breast disease types not strongly associated with breast cancer, all of whom donated blood at the time of diagnosis to the Mayo Serum Bank, a resource established in the 1970's to identify early markers of breast cancer. Results: As hypothesized, mean percent levels of serum DNA methylation were modestly but statistically higher in node-positive breast cancer cases than controls for TWIST (4.9% vs 3.9%; p-value =.002), CyclinD2 (3.2% vs 2.1%; p=.03), and HIN1 (3.9% vs 2.4%; p=.03). In the nine other genes, percent methylation levels were also moderately higher in node-positive cases than controls but did not reach statistical significance (e.g., SRFP1 [6.3% vs 4.3%; p=.06]). For TWIST, Cyclin D2, HIN1, mean percent levels were intermediate in node-negative patients compared to node-positive cases and controls. Additional analyses evaluating the predictive ability of multivariate sets of DNA methylation markers together with established breast cancer risk factors are underway. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 85. doi:10.1158/1538-7445.AM2011-85

Patterns and Risk Factors of Locoregional Recurrence in T1-T2 Node Negative Breast Cancer Patients Treated With Mastectomy: Implications for Postmastectomy Radiotherapy

Postmastectomy radiation therapy (PMRT) can reduce locoregional recurrences (LRR) in high-risk patients, but its role in the treatment of lymph node negative (LN-) breast cancer remains unclear. The aim of this study was to identify a subgroup of T1-T2 breast cancer patients with LN- who might benefit from PMRT. We retrospectively reviewed 1,136 node-negative T1-T2 breast cancer cases treated with mastectomy without PMRT at the Massachusetts General Hospital between 1980 and 2004. We estimated cumulative incidence rates for LRR overall and in specific subgroups, and used Cox proportional hazards models to identify potential risk factors. Median follow-up was 9 years. The 10-year cumulative incidence of LRR was 5.2% (95% CI: 3.9-6.7%). Chest wall was the most common (73%) site of LRR. Tumor size, margin, patient age, systemic therapy, and lymphovascular invasion (LVI) were significantly associated with LRR on multivariate analysis. These five variables were subsequently used as risk factors for stratified analysis. The 10-year cumulative incidence of LRR for patients with no risk factors was 2.0% (95% CI: 0.5-5.2%), whereas the incidence for patients with three or more risk factors was 19.7% (95% CI: 12.2-28.6%). It has been suggested that patients with T1-T2N0 breast cancer who undergo mastectomy represent a favorable group for which PMRT renders little benefit. However, this study suggests that select patients with multiple risk factors including LVI, tumor size ≥2 cm, close or positive margin, age ≤50, and no systemic therapy are at higher risk of LRR and may benefit from PMRT.