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The guidelines Thyroid Cancer 2022 are prepared based on previous Polish recommendations updated in 2018. They consider international guidelines - American Thyroid Association (ATA) 2015 and National Comprehensive Cancer Network (NCCN); however, they are adapted according to the ADAPTE process. The strength of the recommendations and the quality of the scientific evidence are assessed according to the GRADE system and the ATA 2015 and NCCN recommendations. The core of the changes made in the Polish recommendations is the inclusion of international guidelines and the results of those scientific studies that have already proven themselves prospectively. These extensions allow de-escalation of the therapeutic management in low-risk thyroid carcinoma, i.e., enabling active surveillance in papillary microcarcinoma to be chosen alternatively to minimally invasive techniques after agreeing on such management with the patient. Further extensions allow the use of thyroid lobectomy with the isthmus (hemithyroidectomy) in low-risk cancer up to 2 cm in diameter, modification of the indications for postoperative radioiodine treatment toward personalized approach, and clarification of the criteria used during postoperative L-thyroxine treatment. At the same time, the criteria for the preoperative differential diagnosis of nodular goiter in terms of ultrasonography and fine-needle aspiration biopsy have been clarified, and the rules for the histopathological examination of postoperative thyroid material have been updated. New, updated rules for monitoring patients after treatment are also presented. The updated recommendations focus on ensuring the best possible quality of life after thyroid cancer treatment while maintaining the good efficacy of this treatment.

Breast tumors constitute the complex entities composed of cancer cells and stromal components. The compositional heterogeneity should be taken into account in bulk tissue metabolomics studies. The aim of this work was to find the relation between the histological content and 1H HR-MAS (high-resolution magic angle spinning nuclear magnetic resonance) metabolic profiles of the tissue samples excised from the breast tumors and the peritumoral areas in 39 patients diagnosed with invasive breast carcinoma. The total number of the histologically verified specimens was 140. The classification accuracy of the OPLS-DA (Orthogonal Partial Least Squares Discriminant Analysis) model differentiating the cancerous from non-involved samples was 87% (sensitivity of 72.2%, specificity of 92.3%). The metabolic contents of the epithelial and stromal compartments were determined from a linear regression analysis of the levels of the evaluated compounds against the cancer cell fraction in 39 samples composed mainly of cancer cells and intratumoral fibrosis. The correlation coefficients between the levels of several metabolites and a tumor purity were found to be dependent on the tumor grade (I vs II/III). The comparison of the levels of the metabolites in the intratumoral fibrosis (obtained from the extrapolation of the regression lines to 0% cancer content) to those levels in the fibrous connective tissue beyond the tumors revealed a profound metabolic reprogramming in the former tissue. The joint analysis of the metabolic profiles of the stromal and epithelial compartments in the breast tumors contributes to the increased understanding of breast cancer biology.

Optimal therapeutic strategy in low advanced papillary thyroid carcinoma (PTC) is still a matter of debate. The management differs depending on the country.A prospective non-randomized study was performed to evaluate whether less extensive surgery could be a safe, acceptable, and sufficient therapeutic option in PTC cT1N0M0 patients. The present paper summarizes the results of over a 5-year follow-upMaterialOur prospective group (PG) treated between 2011 and 2015 consisted of 139 patients with cT1aN0M0 PTC who underwent lobectomy (LT) as initial surgical treatment (PGcT1aN0M0 group) and 102 cT1bN0M0 patients in whom total thyroidectomy (TT) with unilateral central neck dissection (CND) was performed (PGcT1bN0M0). PG was compared with the retrospective group (RG) of patients who underwent TT with bilateral CND between 2004 and 2006: 103 cT1aN0M0 patients (RGcT1aN0M0) and 91cT1bN0M0 (RGcT1bN0M0). The risks of reoperation, cancer relapse and postoperative complications were analyzed.ResultsOnly 12 cT1aN0M0 patients (7.6%) withdrew from the trial and underwent TT with bilateral CND. Over 90% of patients accepted less extensive surgery. In 4 cT1aN0M0 cases, TT with CND was performed due to lymph node metastases found intraoperatively. The initial clinical stage according to the TNM/AJCC 7th edition was confirmed histologically in 77% of cases in PGT1aN0M0 and in 72% in PGT1bN0M0, respectively. 24 PGcT1aN0M0 patients were reoperated on. In this group, cancer lesions in the postoperative histological specimens were found in 8 cases (32%). Five-year disease-free survival (DFS) was excellent. However, no statistically significant differences were found between PG and RG groups (99.3% in PGcT1aN0M0 and 99.0%, in RGcT1aN0M0; p = 0.41 and 98%, in PGcT1bN0M0 and 94.4% in RGcT1bN0M0; p=0.19). No significant differences were observed in the incidence of early paresis of the recurrent laryngeal nerves between PG and RG. However, as predicted, LT completely eliminated the risk of postoperative hypoparathyroidism.SummaryThe results of the prospective clinical trial confirm that less extensive surgery in adequately selected low-advanced PTC patients is both safe and sufficient.

We investigated the tumor regression grading (TRG) as a prognostic marker for disease-free survival (DFS) in patients with advanced rectal cancer treated within phase III randomized study (ClinicalTrials.gov Identifier: NCT01814969). The study is still recruiting prospective trial of preoperative hyperfractionated radiotherapy (HART) compared with concomitant hyperfractionated radiotherapy with co-administration of chemotherapy based on 5-FU (HART-CT) in patients with T2/N+ or T3/any N resectable rectal cancer. This preplanned interim analysis examined the pathological outcome in the group of 136 patients who were randomly assigned to HART (n = 69) and HART-CT (n = 67). The pelvis was irradiated twice a day (28 fractions of 1.5 Gy), with a minimal interfraction interval of 8 h to total dose 42 Gy over 18 days (HART) or mentioned scheme with concurrent chemotherapy: 5-FU 325 mg/m2 (bolus) on days 1-3 and days 16-18 (HART-CT). Surgery was performed 6-7 weeks after HART/HART-CT. Postoperative 5-FU-based chemotherapy was given to ypN positive patients. The TRG was recorded using the following 4-point scale: TRG0 (pCR) denoted no cancer cells; TRG1 was diagnosed when a few cancer foci had been seen in less than 10% of a tumor mass; TRG2 denoted cancer cells seen in 10-50% of a tumor mass; in order to diagnose TRG3, cancer cells had to be seen in more than 50% of a tumor mass. Multivariable analysis was performed using Cox regression models and Cox proportional hazard model was used in the survival analysis. The crude rate of patients with any serious acute 3 toxicity during the follow-up was 16% vs. 25% for HART and HART-CT. Twenty-two patients (16%) presented with postoperative complications. Anterior resection was performed in 52% vs. 62% for HART and HART-CT respectively (p = 0.06). Of the 136 patients evaluable for pathologic response there were 3 (4%) vs. 9 (13%), 16 (23%) vs. 24 (36%), 40 (58%) vs. 30 (45%), and 10 (15%) vs. 4 (6%) patients with TRG 0, 1, 2, and 3, respectively in HART vs. HART-CT, the difference was statistically significant p = 0.002. The addition of 5-FU infusion to HART was not associated with statistically significant improved loco-regional relapse free survival (LRC), metastasis-free survival (MFS) and DFS. Significant differences in the tumor regression grading (TRG) were found. Both LRC and DFS of rectal cancer patients treated with HART vs. HART-CT had favorable outcomes in the HART-CT arm. Also, the sphincter preservation rate tended to favor HART-CT.

Intra-tumor heterogeneity results from both genetic heterogeneity of cancer (sub)clones and phenotypic plasticity of cancer cells that could be induced by different local microenvironments. Here, we used mass spectrometry imaging (MSI) to compare molecular profiles of primary tumors located in the thyroid gland and their synchronous metastases in regional lymph nodes to analyze phenotypic heterogeneity in papillary thyroid cancer. Two types of cancerous (primary tumor and metastasis) and two types of not cancerous (thyroid gland and lymph node) regions of interest (ROIs) were delineated in postoperative material from 11 patients, then the distribution of tryptic peptides (spectral components) was analyzed by MSI in all tissue regions. Moreover, tryptic peptides identified by shotgun proteomics in corresponding tissue lysates were matched to components detected by MSI to enable their hypothetical protein annotation. Unsupervised segmentation of all cancer ROIs revealed that different clusters dominated in tumor ROIs and metastasis ROIs. The intra-patient similarity between thyroid and tumor ROIs was higher than the intra-patient similarity between tumor and metastasis ROIs. Moreover, the similarity between tumor and its metastasis from the same patients was lower than similarities among tumors and among metastases from different patients (inter-patient similarity was higher for metastasis ROIs than for tumor ROIs). Components differentiating between tumor and its metastases were annotated as proteins involved in the organization of the cytoskeleton and chromatin, as well as proteins involved in immunity-related functions. We concluded that phenotypical heterogeneity between primary tumor and lymph node metastases from the same patient was higher than inter-tumor heterogeneity between primary tumors from different patients.

The study was prospectively designed as a single-arm, single-institution prospective trial of pre-operative concomitant hyperfractionated radiotherapy (HART) with co-administration of chemotherapy based on 5-fluorouracil (5FU) in patients with T2/N+ or T3/any N resectable mid-low primary rectal cancer. The aim of the study was to assess the safety and efficacy of accelerated HART with concurrent 5FU-based chemotherapy in patients with locally advanced rectal cancer.Patients with resectable locally advanced (≥T3 or N+) rectal cancer were eligible. The patients received total dose 42 Gy in 28 fractions of 1.5 Gy, two times daily, with at least 8 h of interval, with concurrent chemotherapy: 325 mg mOut of 53 enrolled patients; 2 did not undergo surgery. Of the 51 patients evaluable for pathological response, there were 8 (15.6%), 20 (39.3%), 18 (35.3%) and 5 (9.8%) patients with tumour regression grade 0, 1, 2 and 3, respectively. Downstaging of the primary tumour and lymph nodes was observed in 22 (43%) and 25 (49%) patients, respectively. The primary tumour ypCR (ypT0) rate was 15% (8/51). The nodal ypCR rate for cN+ patients was 60% (21/35). The total ypCR (ypT0N0M0) rate was 11% (6/51). Toxicity included: Grade 3 diarrhoea (4/51, 7.8%), Grade 2 diarrhoea (22/51, 43.1%), Grade 2 leukopenia (7/51, 13.7%), Grade 2 neutropenia (6/51, 11.7%) and Grade 1 thrombocytopenia (3/51, 5.9%). No Grade 4 toxicity was reported. Nine patients (18%) presented with post-operative complications (during the 3 months after surgery). There were 6 locoregional relapses (11.8%) and distant metastasis occurred in 11 patients (21.6%). The 2-year cumulative locoregional relapse-free survival, metastasis-free survival and overall survival was 87%, 79% and 89%, respectively.The proposed pre-operative HART with co-administration of 5FU had acceptable toxicity profile and provided satisfactory rate of ypCR. This created rationale to initiate a Phase III randomized study that was registered under ClinicalTrials.gov Identifier: NCT01814969. Advances in knowledge: The results of this research show that responders to pre-operative radiochemotherapy have favourable outcome. Tumour regression grade as prognostic clinical feature holds the promise of better classifying patients at high risk of local and systemic recurrence and this issue may be an interesting objective for future research.

Distinguishing between follicular thyroid cancer (FTC) and follicular thyroid adenoma (FTA) constitutes a long-standing diagnostic problem resulting in equivocal histopathological diagnoses. There is therefore a need for additional molecular markers. To identify molecular differences between FTC and FTA, we analyzed the gene expression microarray data of 52 follicular neoplasms. We also performed a meta-analysis involving 14 studies employing high throughput methods (365 follicular neoplasms analyzed). Based on these two analyses, we selected 18 genes differentially expressed between FTA and FTC. We validated them by quantitative real-time polymerase chain reaction (qRT-PCR) in an independent set of 71 follicular neoplasms from formaldehyde-fixed paraffin embedded (FFPE) tissue material. We confirmed differential expression for 7 genes (CPQ, PLVAP, TFF3, ACVRL1, ZFYVE21, FAM189A2, and CLEC3B). Finally, we created a classifier that distinguished between FTC and FTA with an accuracy of 78%, sensitivity of 76%, and specificity of 80%, based on the expression of 4 genes (CPQ, PLVAP, TFF3, ACVRL1). In our study, we have demonstrated that meta-analysis is a valuable method for selecting possible molecular markers. Based on our results, we conclude that there might exist a plausible limit of gene classifier accuracy of approximately 80%, when follicular tumors are discriminated based on formalin-fixed postoperative material.

Introduction. Despite the use of multiple diagnostic imaging techniques, a non-invasive differential diagnosis of focal lesions in the liver is still a challenging task. It is even more significant, considering the risk of complications related to liver biopsy. On the other hand, the availability of non-invasive radiological and diagnostic imaging methods has resulted in the increased detectability of incidental hepatic lesions. Therefore, further investigation of newer and more effective methods of differentiating benign from malignant hepatic lesions is extremely useful. The aim of the study was to assess the diagnostic value of Diffusion-Weighted Magnetic Resonance Imaging (DW MRI) in differentiating benign from malignant focal lesions in the liver. Material and methods. Fifty one patients with hepatic lesions detected by magnetic resonance (MR) examination were enrolled in the study. Each lesion was then verified histopathologically, cytologically, or by observation. Ninety five (71 malignant and 24 benign) hepatic focal lesions were analysed. Apart from a standard MR liver examination, an echo planar imaging spin echo diffusion weighted sequence was performed, with a slice thickness of 5 or 6 mm. The maximal b-value was 1000 s/mm 2 and the minimal 0 s/mm 2 .In the study, the following were assessed: the presence of restricted diffusion, an absolute ADC (apparent diffusion coefficient) value, the decrease of signal intensity in DW MRI with an increase in the b-value. Results. Confirming or ruling out the restricted diffusion is usually the first effective step in differentiating benign from malignant hepatic lesions, as indicated in the study results. In the material analysed, the diffusion restriction occurred statistically significantly more frequently in malignant (87.3 %) compared to benign lesions (12.5%), (p 1.04 × 10 -3 mm2/s to be benign and lesions with ADC value ≤ 1.04 × 10 -3 mm 2 /s to be malignant, 75.8% lesions were correctly classified. Conclusions. The results obtained indicate that restricted diffusion, as a marker for malignant lesions, has the highest value in DW MRI in differentiating benign from malignant lesions in the liver.

Wstep: Rak pecherzykowy tarczycy (FTC) jest nowotworem ktorego podloze molekularne jest malo zbadane. W podjetej analizie transkryptomu oceniono mozliwośc dyskryminacji raka i gruczolaka pecherzykowego tarczycy (FTA) na podstawie badan profilu ekspresji genow metodą tzw. nienadzorowaną (tzn. na podstawie dominujących źrodel zmienności). Analize te prowadzono by sprawdzic czy zlośliwośc guza jest rzeczywiście czynnikiem dominującym dla profilu ekspresji genow w nowotworach pecherzykowych. Material i metody: Podstawowy zbior guzow pecherzykowych obejmowal 52 probki (27 FTC i 25 FTA), z ktorych wyizolowano calkowity RNA i poddano badaniu na mikromacierzach HG-U133 Plus 2.0. Otrzymany zbior normalizowano za pomocą RMA i GC-RMA. Identyfikacji glownych źrodel zmienności dokonano metodą analizy glownych skladowych (PCA). Wyniki: Analize funkcji biologicznej genow przeprowadzono dla pierwszych 6 skladowych glownych. Geny skorelowane z pierwszą skladową pozwalaly wyodrebnic 2 klastry probek: jeden zlozony glownie z gruczolakow, z wysoką ekspresją miedzy innymi transkryptow tarczycowo-swoistych, drugi zaś, zawierający wiekszośc rakow, wykazywal zwiekszoną, ale heterogenną ekspresje genow związanych z odpowiedzią immunologiczną, a obnizoną ekspresje genow tarczycowych. Geny odpowiedzi immunologicznej stwierdzono wśrod transkryptow skorelowanych przebiegiem pierwszej, trzeciej i szostej glownej skladowej; w istotny sposob wplywaly one na rozroznienie miedzy FTC i FTA. Wnioski: W analizie nienadzorowanej stwierdzono, ze zlośliwośc (inwazyjnośc) nowotworu pecherzykowego moze byc jednym z glownych źrodel zmienności w transkryptomie tych guzow. Jednak, genomiczna odleglośc miedzy grupami FTC i FTA jest niewielka, a wyodrebnione w analizie nienadzorowanej klastry nakladają sie, stąd sama analiza nienadzorowana nie jest wystarczającym narzedziem do celow klasyfikacji tych guzow. (Endokrynol Pol 2013; 64 (5): 329–334)

The inherent diagnostic limitations of thyroid fine needle aspiration (FNA), especially in the "indeterminate" category, can be partially overcome by molecular analyses. We aimed at the identification of miRNAs that could be used to improve the discrimination of indeterminate FNAs. miRNA expression profiling was performed for 17 follicular carcinomas (FTCs) and 8 follicular adenomas (FAs). The microarray results underwent cross-comparison using three additional microarray data sets. Candidate miRNAs were validated by qPCR in an independent set of 32 FTCs and 46 FAs. Sixty-eight differentially expressed miRNAs were identified. Thirteen miRNAs could be confirmed by cross comparison. A two-miRNA-classifier was established improving the diagnostic applicability and resulted in a sensitivity of 82% and a specificity of 49%. We present a classifier that has the potential to be successfully evaluated in cytology material for its capability to discriminate (mutation negative) indeterminate cytologies and thereby improving the pre-surgical diagnostics of thyroid nodules.

neal cylindromatosis was presented. A young woman was admitted to hospital after removal of a right ovarian cystic tumour. Multiple histological reexaminations suggested the presence of a dermoid cyst, i.e. adult teratoma ad ditionally with granulosa cell tumour and endometrioid cancer. Six years after complete removal of neoplastic foci, a routine examination revealed multiple nodular lesions inside the pelvis and in the pelvic peritoneum. Multiple histological studies confirmed the presence of a cylindroma. Based on the broad histochemistry and immunohistochemistry (periodic acid Shiff, Ber-EP4, p63, CKAE1/AE3, CEA positive, as opposed to negative results with inhibin, calretinin, CK7 and only focally positive for EMA; Ki 67 mitotic index did not exceed 5% of cells), granulosa cell tumour and endometrioid cancer was excluded. A comparison of the surgical material with the specimens taken from the primary lesion operated eight years earlier showed a similarity of histological images. Our case of peritoneal dissemination of eccrine cylindroma arising from dermoid cyst suggests the introduction of the term peritoneal cylindromatosis for similar cases.

BACKGROUND: The search for new surgical flap techniques and modifications of already existing ones is gaining increasing popularity. Progress in flap designing and harvesting have improved the functional and aesthetic results, especially in head and neck reconstruction. MATERIAL/METHODS: Ten pigs were used in this study. In the first operation, high-density porous polyethylene prefabrication was performed bilaterally in all pigs. After 8 weeks, each prefabricated complex was explored, resected, and macroscopically evaluated. RESULTS: All of 20 prefabricated flaps survived. No serious surgical complications were observed. In 2 cases there was chronic inflammation and in 4 cases there was instability of the implant. CONCLUSIONS: After this experimental study, we believe that the use of high-density porous polyethylene in flap prefabrication may be a good option for reconstruction of 3-dimensional defects, especially in patients with limited donor tissues.

Genetic alterations involving the mitogen-activated protein kinase (MAPK) pathway are frequently demonstrated in papillary thyroid cancer (PTC). BRAF(V600E), the most frequent mutation in adult patients, is present in approximately 50% of PTC. Most clinical studies have demonstrated an association of BRAF(V600E) mutation with aggressive clinicopathological characteristics and high tumour recurrence, although the results are controversial. In this study we present the preliminary results of BRAF mutation frequence in a group of 88 Polish patients with papillary thyroid cancer (PTC) and relate it to the outcome all DTC patients operated in 2004 and 2005. BRAF (V600E) mutation was diagnosed in 38 (43%) of cases.The presence of BRAF mutation was evaluated in 88 PTC tumours. DNA was isolated from tissue parafin blocks, and the mutation V600E was evaluated by sequence analysis with an AbiPrism 377 and 3130 xl genetic analyzer (Life Technologies). Statistical analysis was carried out with the use of SPSS 12 software. The chi² and Kaplan-Meyer survival analysis were performed.From all analyzed clinico-pathological factors, only older age positively correlated with BRAF mutation frequency (p = 0.0017). Lymph node/distant metastases, multifocality, and extra-thyroid extension did not correlate with BRAF status. One cancer related death and two reccurences were observed in the BRAF+ group while one relapse was diagnosed in the BRAF- group.Although many studies document BRAF mutation as a prognostic factor in PTC our results underline that it is too early to consider it as a routine clinical predictive factor.

In differentiated thyroid carcinoma (DTC) with primary tumor smaller than 1 cm, the routine central lymph node (LN) dissection is questioned, due to increased risk of post-surgery complications and lack of confirmed benefit. Aim The analysis of prognostic significance of LN metastases, in DTC patients to verify the potential role of central neck lymphadenectomy on disease staging. Materials and methods The group of 195 DTC patients, primarily operated between 2004 and 2005, was retrospectively analyzed. 184 patients after radical operation, with no distant metastases diagnosed before surgery, were included into analysis. LN metastases were observed in 55 of cases (28%). In 124 cases only dissection of central LN compartment was performed, in 36 patients also uni- or bilateral modified cervical lymphadectomy was carried out. In 24 patients with tumor limited to the thyroid gland without suspicious lymph nodes, the routine central lymph node dissection was not done. Results Median follow-up was 4 years. The 5-year overall and disease free survival standardized ratio were 100% and 95% respectively. The risk of LN metastases increased with the more locally advanced cancer. In the group of 124 patients, in whom only central LN dissection was performed, LN metastases were diagnosed in 15 cases (12%). No significant relation between multifocality and frequency of central and/or lateral LN metastases was noticed. Significant correlation between N feature and extrathyroidal invasion was observed (p = 0,0003). The presence of LN metastases was related to worsening of disease free survival from 99 to 90%. During the follow-up recurrence occurred in 6 (3%) cases. In 24 patients in whom only total thyroidectomy was done, no local or distant recurrence was observed. The assessment of early postoperative complications (hypoparathyroidism, paresis of vocal cords) indicated that the frequency of early calcium balance disturbances was significantly lower in patients in whom central LN dissection was not performed (p = 0,04) Conclusions Our result indicate that in the early diagnosis of thyroid cancer, the occurrence of LN DTC metastases is rarer and was observed only in 12% of elective dissections of central LN node compartment, if no lateral dissection was indicated due to the lack of clinical suspicion. In DTC patients with tumor diameter

Introduction The risk of over-treatment in low-advanced PTC stages has prompted clinicians to search for new reliable prognostic factors. The presence of BRAF mutation, the most frequent molecular event in PTC, seems to be a good candidate. However, there is still lack of randomised trials and its significance has been proved by retrospective analyses, involving a large group of patients. The question arises whether this factor is useful in smaller populations, characterised for specialised centres. Thus, the aim of the study was to evaluate the use of BRAF mutation as a potential predictive marker in PTC patients. Material 233 PTC subjects treated between 2004-2006, were retrospectively analysed. Stage pT1 was diagnosed in 64.8% patients and lymph node metastases in 30.9%. Median follow-up was 7.5 years. BRAFV600E mutation was assessed postoperatively in all cases. Results BRAF V600E mutation was found in 54.5%. It was more frequent in patients > 45 years (p=0.0001), and associated with larger tumour size (p=0.004). Patients with tumours

Background Differential diagnosis between malignant follicular thyroid cancer (FTC) and benign follicular thyroid adenoma (FTA) is a great challenge for even an experienced pathologist and requires special effort. Molecular markers may potentially support a differential diagnosis between FTC and FTA in postoperative specimens. The purpose of this study was to derive molecular support for differential post-operative diagnosis, in the form of a simple multigene mRNA-based classifier that would differentiate between FTC and FTA tissue samples. Methods A molecular classifier was created based on a combined analysis of two microarray datasets (using 66 thyroid samples). The performance of the classifier was assessed using an independent dataset comprising 71 formalin-fixed paraffin-embedded (FFPE) samples (31 FTC and 40 FTA), which were analysed by quantitative real-time PCR (qPCR). In addition, three other microarray datasets (62 samples) were used to confirm the utility of the classifier. Results Five of 8 genes selected from training datasets (ELMO1, EMCN, ITIH5, KCNAB1, SLCO2A1) were amplified by qPCR in FFPE material from an independent sample set. Three other genes did not amplify in FFPE material, probably due to low abundance. All 5 analysed genes were downregulated in FTC compared to FTA. The sensitivity and specificity of the 5-gene classifier tested on the FFPE dataset were 71% and 72%, respectively. Conclusions The proposed approach could support histopathological examination: 5-gene classifier may aid in molecular discrimination between FTC and FTA in FFPE material.

556 Background: The aim of the study is to assess radiation-induced tumor and normal mucosa responses in patients who underwent preoperative radiotherapy according to two different fractionation protocols. This research constitutes one of the secondary endpoints of a prospective randomized trial. Methods: Pathological samples after preoperative radiotherapy and surgery were obtained from 106 patients: 50 were treated with hypofractionation 39 Gy in 3.0 Gy per fraction (HYPO) and 56 with accelerated hyperfractionation 42 Gy given in two daily fractions of 1.5 Gy with a minimal interfraction interval of 6 hours (HART). This sample represents approximately 30% of an overall trial size. Over 50 pathological features were assessed for each sample including tumor/nodal response, normal mucosa and cellular matrix response, ulceration and morphology of tumor cells. The observed grades of responses in HYPO vs. HART were compared using U Mann-Whitney test. Results: The primary tumor downstaging was observed in 41% vs. 35% of the patients treated with HYPO and HART respectively, the difference was not significant (p=0.5). By contrast, a trend towards improved nodal response among the patients treated with HYPO was recorded, with 94% achieving complete or major nodal response, compared to 73% in HART (p=0.03). Mucosal atrophy and edema were significantly higher in HYPO (p=0.003 and p=0.02, respectively). A trend for increased mucosal inflammation was also recorded in HYPO (p=0.06). Tumor/nodal responses were not significantly influenced by time-interval radiotherapy-surgery (below or above the median of 11 days), however, more cellular injury to normal mucosa (atrophy, ulceration) was observed at short RT-surgery intervals (p=0.01). Conclusions: Radiation-induced tumor and normal mucosa responses differ significantly according to fractionation schedule, with more mucosa injury observed in HYPO. Pathological responses of mucosa are also influenced by time-interval radiotherapy-surgery.

506 Background: The aim of the study is to verify hypothesis that accelerated hyperfractionated preoperative radiotherapy for rectal cancer (HART) may provide a favorable long-term tolerance compared to treatment given in higher fraction (fx) doses (HYPO). This report focuses on early outcomes of the study. Comparison of late tissue reactions, evaluation of QLQ and long-term outcome in both trial arms is the ultimate goal. Methods: Between 2005 and 2011, 238 patients (pts) with cT3-4 resectable adenocarcinoma of the rectum were enrolled, which represents app. 70% of an overall trial size. The pts were randomly assigned to HART (n=122) or HYPO (n=116). The pelvis was irradiated 2x/day to the total dose of 42 Gy in 1.5 Gy/fx over 18 days (HART). Patients in HYPO received 39 Gy in 3.0 Gy/fx over 17 days. Postoperative chemotherapy (PCT) was given to ypN+ pts. Results: The actuarial perioperative complication rate at 1 year after treatment was 24.4% vs. 33.4% for HART and HYPO resp. (RR=0.69, p=0.13). The crude rate of pts with any surgical complication during the follow-up (median of 3.1 year) was 32/122 (26.2%) vs. 36/116 (31.0%) for HART and HYPO resp. (p= 0.41). If the values were expr. In terms of number of events (some pts had more than one complication) the rates were 55 events per 122 pts (HART) vs. 63 events per 116 pts (HYPO). Anastomotic leakage appeared in 11/122 HART pts (9.0%) vs. 16/116 in HYPO (13.8%). Anterior resection was performed in 48.8 % vs. 44.1% for HART and HYPO resp. PCT was given to 46.1% of the pts in HART and 45.4% in HYPO. There were no significant differences in 3-year loco-regional control (RR=1.08, p=0.44) and overall survival (RR=0.97, p=0.72) between HART and HYPO, with actuarial rates (for both arms) of 85% and 73% resp. Neither the rate of complicatons, nor type of surgery were influenced by duration of the interval RT-surgery (median 11, Std±18 days) Conclusions: The perioperative complication rate observed had the propensity to favor HART over HYPO, but the differences did not reach the level of statistical significance. Longer follow-up and the analysis encompassing all of the pts enrolled is required to fully address the study endpoints.

e14159 Background: Chemotherapy is the mainstay of treatment patients with metastatic colorectal cancer. The choice of first-line treatment is difficult, especially when cost-effectiveness is the primary constraint. Thus, the optimal use of the clinical and biological factors influencing prognosis would be beneficial. The aim of our study was to identify factors affecting the time to progression (TTP) after first-line FOLFOX chemotherapy in palliative setting. Methods: The study is a retrospective analysis of the series of consecutive patients from large cancer center in south of Poland. The analysis was carried out in the group of 180 patients (37.2% of women), treated between 2007-2010 by FOLFOX-4 regimen and followed-up with the median time of observation 16.3 month. Patients received chemotherapy with median time of 5.0 months, median 10 cycles. Progression was defined as PD by RECIST criteria, death due to disease or sympomatic deterioration. 94 paraffin blocks were available for KRAS testing and gene expression analysis by real-time PCR. Results: The median TTP (counted from beginning of chemotherapy) was 8.6 month, the median TTP from the end of treatment was 3.4 month. We tested the wide range of clinical variables associated with both disease and and patient status by multivariate Cox regression analysis. Two most potent independent negative predictors were identified: the presence of massive lymph node involvement as assessed in CT scan before palliative treatment (>10 nodes enlarged) – hazard ratio 2.82, p

e21049 Background: Gene expression profiling has been shown to be effective in the analysis of postoperative tumor samples in breast cancer. However, when small specimens like core biopsy samples are considered, the small size of the collected material makes it difficult to obtain high tumor percentage in the specimen. Thus, we tested the strategy of obtaining three independent cores for molecular analysis and assessed the influence of the stability of different prognostic and predictive signatures. Methods: We prospectively collected breast core biopsies from 82 consecutive patients treated by pre-operative chemotherapy. In each patient, three cores from different tumor areas were sampled for molecular testing, independent of the cores collected for histopathological analysis. RNA was isolated by QIAGEN RNeasy kits, gene expression profiling was carried out using Affymetrix HG-U133 Plus 2.0 microarrays. Results: In 27 patients, three independent specimens were analyzed by microarray gene expression profiling (in total 81 samples). When this training set was analyzed by unsupervised Principal Component Analysis, the overall concordance between the three tumor cores was observed, but no such concordance was seen for three tumors (for each with one significant outlier). When we assessed the set of chosen 20 prognostic and predictive signatures, we confirmed that the homogeneity of gene expression within the signatures is significantly better than for random sets of genes, but we observed the notable differences between the signatures. The heterogeneity decreased when genes with low between-patient variance were excluded; they also showed a higher intra-patient variability of gene expression. We also assessed the heterogeneity of single expression markers and furher validated our findings on an independent set of 55 patients. Conclusions: The use of three independent cores for gene expression testing in breast cancer may be a successful strategy to overcome tumor heterogeneity and sampling error and result in more stable results of prognostic and/or predictive signatures. Supported by Polish National Science Center grant N402 6861 40

There is an ongoing debate whether hereditary breast cancer is a clinical entity distinct from sporadic breast cancer. We tried to shed some light on this issue by comparing the molecular profiles of these two types of cancer using DNA microarrays. Our results show that a previously reported marked difference between BRCA1-mutation linked and sporadic breast cancer was probably due to uneven stratification of samples with different ER status and basal-like versus luminal-like subtype. We observed that apparent difference between BRCA1-linked and other types of breast cancer found in univariate analysis was diminished when data were corrected for ER status and molecular subtype in multivariate analyses. In fact, the difference in gene expression pattern of BRCA1-mutated and sporadic cancer is very discrete. These conclusions were supported by the results of Q-PCR validation. We also found that BRCA1 promoter hypermethylation had similar effect on global gene expression as mutation-induced protein truncation. Thus, in the molecular studies of hereditary breast cancer, BRCA1 promoter methylation should be recognized and considered together with gene mutation.

Global analysis of gene expression by DNA microarrays is nowadays a widely used tool, especially relevant for cancer research. It helps the understanding of complex biology of cancer tissue, allows identification of novel molecular markers, reveals previously unknown molecular subtypes of cancer that differ by clinical features like drug susceptibility or general prognosis. Our aim was to compare gene expression profiles in breast cancer that develop against a background of inherited predisposing mutations versus sporadic breast cancer. In this preliminary study we analysed seven hereditary, BRCA1 mutation-linked breast cancer tissues and seven sporadic cases that were carefully matched by histopathology and ER status. Additionally, we analysed 6 samples of normal breast tissue. We found that while the difference in gene expression profiles between tumour tissue and normal breast can be easily recognized by unsupervised algorithms, the difference between those two types of tumours is more discrete. However, by supervised methods of data analysis, we were able to select a set of genes that may differentiate between hereditary and sporadic tumours. The most significant difference concerns genes that code for proteins engaged in regulation of transcription, cellular metabolism, signalling, proliferation and cell death. Microarray results for chosen genes (TOB1, SEPHS2) were validated by real-time RT-PCR.