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Background:Triple-negative breast cancer (TNBC) has the worst prognosis and the highest immunogenic potential of all breast cancer subtypes. The tumour microenvironment (TME) of TNBC consists mostly of tumour-infiltrating lymphocytes (TILs), tumour-associated macrophages and dendritic cells. TILs are involved in host immunity against tumour cells through the activation of tumour-specific CD8+ cytotoxic T cells. However, there are opposing data about the prognostic value of TILs in TNBC. Programmed cell death receptor ligand 1 (PD-L1) from immune or tumour cells binding programmed cell death receptor 1 (PD-1) disable the effector role of CD8 T cells. Therefore, antibodies that block the target in the PD-1 signalling pathway elicit a stronger immune response. Cytotoxic T lymphocyte-associated protein 4 (CTLA-4) mediates immunosuppression and it is expressed in tumours on infiltrating Tregs, activated CD4+ T cells, exhausted T cells and tumour cells. In tumours with high TILs, PD-L1 and CTLA-4 blockades are more effective. Methods:We have performed a comprehensive IHC analysis of all major TIL components (CD8, CD4, FOXP3 Tregs) as well as inhibitory molecules PD-L1 and CTLA4) in a superficial (invasive tumour front, ITF) and deep tumour layer of TNBC, and compared it with established clinicopathological and prognostic parameters. Clinical data and surgical tissue samples from 68 TNBC patients who underwent initial surgery were included in the analysis and 36 control samples from benign breast tissue biopsies. Results:Several statistically significant associations between the TILs status of TNBC patients and the established prognostic factors were observed. In the ITF, the proportion of TILs and CD8+T cells were increasing toward second pathological T status (pT2), and decreasing thereafter toward higher pT status (P=0.017, P=0.021, Chi-square test). Similar trends for both variables were observed in association with anatomic (P=0.057, P=0.050, Chi- square test) and prognostic (P=0.059, P=0.048, Chi-square test) stages of the disease. Furthermore, the increase of CD8+T cells at ITF was statistically correlated with the increased expression of PDL-1, CTLA-4, FOXP3 and CD4+T cells (N=65, rho 0.31, P=0.011 ; N=65, rho 0.40, P

Rad govori o osnovanju centra za liječenje raka dojke sa europskim certifikatom.

Metallothioneins (MTs) are a family of small cysteine-rich proteins involved in many physiological and pathological processes. Genes that encode the four isoforms of metallothioneins (MT1-M4) are located on chromosome 16q13. Strucutures of these four isoforms allow metallothioneins their various biological functions. Many studies have shown that MT plays an important role in carcinogenesis, tumour growth, its progression from local to metastatic disease and may contribute to resistence to chemotherapy and radiotherapy. Due to the fact that breast cancer is one of the leading causes of death in women worldwide it is important to better understand the biology of breast cancer. So, findings of MT could evenutally help as a prognostic tool and could lead to a possible new specific anti-cancer treatment., Metalotioneini su skupina proteina bogatih cisteinima koji su uključeni u mnoge fiziološke i patološke procese. Geni koji kodiraju četiri izoforme metalotioneina locirani su na lokusu 16q13. Strukture ovih četiriju izoformi omogućuju metalotioneinima njihove razne biološke funkcije. Mnoge su studije pokazale da metalnotionein ima važnu ulogu u karcinogenezi, rastu tumora, njegovoj progresiji od lokalne prema metastatskoj bolesti te je povezan sa razvojem rezistencije na kemoterapiju i radioterapiju. Rak dojke jedan je od vodećih uzroka smrti u svijetu i važno je bolje razumijeti ulogu metalotioneina u različitim podtipovima karcinoma dojke. Ovakvi podaci mogli bi pomoći kao prognostički alat i voditi pronalasku novog specifičnog liječenja karcinoma.

Rad govori o kirurškom pristupu karcinomu dojke u ranom stadiju bolesti.

Background: Our objective was to assess the effects of COVID-19 antiepidemic measures and subsequent changes in the function of the health care system on the number of newly diagnosed breast cancers in the Republic of Croatia. Subjects, materials, and methods: We performed a retrospective, population- and registry-based study during 2020. The comparator was the number of patients newly diagnosed with breast cancer during 2017, 2018, and 2019. The outcome was the change in number of newly diagnosed breast cancer cases. Results: The average monthly percent change after the initial lockdown measures were introduced was -11.0% (95% confidence interval - 22.0% to 1.5%), resulting in a 24% reduction of the newly diagnosed breast cancer cases in Croatia during April, May, and June compared with the same period of 2019. However, during 2020, only 1% fewer new cases were detected than in 2019, or 6% fewer than what would be expected based on the linear trend during 2017-2019. Conclusion: It seems that national health care system measures for controlling the spread of COVID-19 had a detrimental effect on the number of newly diagnosed breast cancer cases in Croatia during the first lockdown. As it is not plausible to expect an epidemiological change to occur at the same time, this may result in later diagnosis, later initiation of treatment, and less favorable outcomes in the future. However, the effect weakened after the first lockdown and COVID-19 control measures were relaxed, and it has not reoccurred during the second COVID-19 wave. Although the COVID-19 lockdown affected the number of newly diagnosed breast cancers, the oncology health care system has shown resilience and compensated for these effects by the end of 2020. Implications for practice: It is possible to compensate for the adverse effects of COVID-19 pandemic control measures on breast cancer diagnosis relatively promptly, and it is of crucial importance to do it as soon as possible. Moreover, as shown by this study's results on the number of newly diagnosed breast cancer cases during the second wave of the pandemic, these adverse effects are preventable to a non-negligible extent.

Aim of the study: Membrane EGFR (mEGFR) protein overexpression is frequently found in the head and neck squamous cell carcinoma (HNSCC). It has been found that mEGFR upon stimulation translocates to nucleus and its nuclear localisation is associated with poor prognosis in many cancers. The main focus of this study is to asses if nuclear EGFR (nEGFR) expression affects biologically more aggressive tumor behaviour in comparison to mEGFR expression in laryngeal SCC. Material and Methods: We examined 42 laryngeal squamous cell carcinomas (SCC) for nEGFR and mEGFR expression as well as cell cycle proliferative markers Ki-67, p53, cyclin D1 using immunohistochemistry. Results: In our study group, we found in 28.57% (12/42) SCC cases a strong (3+) nEGFR expression, 64.28% (27/42) SCC had weak to moderate (1+/2+) nEGFR expression while 7.14% (3/42) cases were negative for nEGFR. The majority of patients with SCC had strong (3+) mEGFR (52.38% or 22/42) expression and 45.23% (19/42) had weak to moderate (1+/2+) mEGFR expression and one case (1/42) was negative for mEGFR. The mean values ± standard deviation (%) of Ki-67, p53 and cyclin D1 expression in our study group were 39.04 ± 18.08, 38.88 ± 32.22 and 43.82 ± 18.34, respectively. When assessing the association of nEGFR with mEGFR and cell cycle proliferation markers there was statistically significant negative correlation between nEGFR and mEGFR expression (τ = -0.389 ; P = 0.002) and statistically significant negative correlation between nEGFR and cyclin D1 (τ = -0.274 ; P = 0.032). In the analysis of mEGFR correlations with the examined cell proliferation markers there were no statistically significant associations. We also observed that higher number of patients with strong nEGFR expression and concomitant negative/weak to moderate mEGFR expression died (70% or 7/10 patients) in comparison to number of patients with strong mEGFR expression and negative/weak to moderate nEGFR expression (40% or 8/20 patients). Moreover, univariate statistical analysis showed a statistically significant correlation between strong nEGFR protein expression with worse overall survival in laryngeal SCC, alone or in co-expression with strong cyclin D1 and high Ki-67 (P=0.025, P=0.046, P=0.043, respectively). However, there was no statistically significant difference in the overall survival, when we analyzed strong mEGFR expression, alone or in co- expression with cyclin D1 and Ki-67 cell cycle proteins (P=0.953, P=0.731, P=0.647, respectively). Conclusion: Our data indicate that nuclear EGFR cellular localization with strong expression might influence the more aggressive biological behaviour of laryngeal SCC carcinoma with poor patient survival.

There are only a few studies so far on IMP3 role as a potential diagnostic biomarker in the laryngeal carcinogenesis. Therefore, it makes sense to analyze IMP3 together with other biomarkers of carcinogenesis. The aim of this study was to investigate the immunohistochemical expression patterns of IMP3, Ki-67, p53 and cyclin D1 in laryngeal carcinogenesis. The study included 153 patients divided into three groups: 68 operated for primary invasive laryngeal squamous cell carcinoma (LSCC) ; 41 with precancerous lesions of atypical and abnormal hyperplasia and 44 with hyperplastic laryngeal nodule without atypia. Tissue microarray technique was used for immunohistochemical analysis. As for the IMP3 staining pattern, a cytoplasmic staining of low intensity in a few cells of superficial layers was found in both control and precancerosis groups. On the other hand, in tumor cells of LSCC there was a high percentage of cytoplasmic staining against IMP3 of high intensity. Additionally, IMP3 staining showed significant intertumoral and intratumoral heterogeneity in different cases of LSCC. In well- differentiated LSCC with present keratinization we noticed that negative areas of the tumor alternate with areas of cytoplasmic staining of low intensity. In tumor cells of moderately differentiated LSCC there was a pattern of combined focal cytoplasmic staining of strong intensity with cytoplasmic staining of low intensity, while a diffuse cytoplasmic staining of moderate to strong intensity was found in tumor cells of poorly differentiated LSCC. Ki67 staining showed a low percentage of nuclear staining of medium to high intensity against Ki-67 in parabasal and a few suprabasal layers of samples from both control and precancerosis groups. High percentage of nuclear staining of high intensity was found in samples from the LSCC group. p53 staining showed different patterns in all three groups. In control group, there was a low percentage of nuclear staining of moderate to strong intensity in parabasal and few suprabasal layers, while in precancerosis group it was medium percentage of nuclear staining of moderate to strong intensity in basal, parabasal and few suprabasal layers. In the LSCC group there was a high percentage of nuclear staining of moderate to strong intensity in tumor cells. Cyclin D1 showed a high percentage of nuclear staining of moderate to strong intensity in parabasal and few suprabasal layers of samples from the control group and in tumor cells of LSCC. However, in the precancerosis group there was a medium percentage of nuclear staining of moderate to strong intensity in a few basal, parabasal and several suprabasal layers. Immunohistochemical expressions of Ki-67 and pronouncedly IMP3 generally follow the same pattern where control and precancerosis are similar and LSCC significantly differs, as opposed to p53 and cyclin D1. In that sense, IMP3 expression increase and difference in LSCC, as opposed to control and precancerosis, is especially pronounced, which points toward its possibly important diagnostic, therapeutic and prognostic value.Further studies on the exact molecular mechanisms behind these differences are, of course, needed.

Cilj istraživanja je bio utvrditi utjecaj pandemije COVID-19 na sveukupni rad patohistološkog i citološkog laboratorija u KBC-u Rijeka kao i na dijagnostiku nekih od najčešćih malignih bolesti. Retrospektivno su iz računalne baze podataka izvučeni i međusobno uspoređeni brojevi patohistoloških i citoloških nalaza, novodijagnosticiranih malignoma dojke, pluća, debelog crijeva, endometrija i prostate te molekularnih testova za analizu mutacije EGFR i HPV-infekcije za razdoblje od 1. ožujka do 30. rujna 2019. (prije pandemije) te od 1. ožujka do 30. rujna 2020. (za vrijeme pandemije COVID-19). Ukupni broj izvršenih pretraga bio je statistički značajno manji za vrijeme pandemije COVID-19 u 2020. u odnosu na isto razdoblje 2019. (p

INTRODUCTION: Breast cancer is one of the leading causes of cancer related deaths. Immune infiltration of breast tumors has been shown to be related to clinical outcome. Natural killer (NK) cells are effector lymphocytes involved in tumor immunosurveillance. In solid malignancies, tumorassociated NK cells in peripheral blood and tumor-infiltrating NK cells show altered phenotypes and are characterized by either anergy or reduced cytotoxicity. The goal of study was to investigate the immune activation profile of NK cells in order to gain a better understanding of pathological behaviour of different breast cancer subtypes. MATERIAL AND METHODS: Immunohistology was used to detect presence and localization of CD56 and IL-15 in paraffin embedded normal and tumoral breast tissue sections. The distribution and frequency of NKG2A, NKG2C, NKp46, CD94, CD69 and CD107a, was investigated in population of NK cells in mononuclear cell suspensions from peripheral blood by flow cytometry. Cytolitic mediatorꞌs mRNA was detected by quantitative RT- qPCR. RESULTS: The percentage of IL15+ and CD56+ cells were significantly higher in triple negative breast cancer tissue. The frequency of NK cell activating receptors were decreased in breast cancer subtypes while inhibitory receptor NKG2A was increased. Decreased percentage of CD69+ /CD107a+ NK cell population indicated lower cellular function and cytotoxicity. Gene expression of cytolitic mediators at local level were up regulated in luminal B breast cancer. CONCLUSION: Modulation of NK cells activity in peripheral blood and tumor-infiltrating NK could be involved in pathogenesis of breast cancer. This highlights the importance of NK cells as a suitable target to modulate the immunosuppressive tumour microenvironment and try to trigger a more potent anti-tumor response. Acknowledgement: The experiments were financed by the grant No. 19-171498

Uvod: Papilarni karcinom je najčešći karcinom štitne žlijezde koji ima tendenciju metastaziranja u limfne čvorove vrata gdje se uglavnom rano dijagnosticira ultrazvukom. Rijetki su slučajevi kada se pronađu metastatski čvorovi u vratu, a da primarni tumor u štitnjači nije moguće potvrditi. Cilj ovog rada je prikazati slučaj metastatskog papilarnog karcinoma porijekla folikularnih stanica štitnjače koji je dijagnosticiran jedino u limfnim čvorovima vrata, a niti nakon opsežne patohistološke analize nije potvrđen u štitnoj žlijezdi. Prikaz slučaja: Pedesetpetogodišnji pacijent primijetio je unatrag mjesec dana bezbolnu oteklinu na vratu. Učinjen je ultrazvuk vrata pri čemu je otkrivena cistična lezija na desnoj strani vrata, u regiji III te je postavljena sumnja na sekundarizam koja citološkom punkcijom nije potvrđena. CT-om vratnih organa uočeni su patološki limfni čvorovi u regijama III i IV i VB desne strane vrata kao i uvećanje desne palatalne tonzile. Patohistološki nalaz tkiva desne tonzile je opisao subkronični limfadenitis dok je nalaz limfnog čvora morfološki i imunohistokemijski govorio u prilog metastaze papilarnog karcinoma štitne žlijezde. U preparatima limfnih čvorova disektata desne strane vrata patohistološki je pronađeno šest metastatskih limfnih čvorova (1 u regiji II, 3 u regiji III, 1 u regiji IV i 1 u regiji VB) od kojih je najveći u regiji IV bio cistično promijenjen. Epitelne stanice nisu pokazivale jasne nuklearne karakteristike papilarnog karcinoma dok je imunohistokemijsko bojenje na Thyreoglobulin, TTF i CK19 bilo snažno i difuzno. U tkivu štitne žlijezde, koje je ekstenzivno uzorčeno, nije pronađeno tumorsko tkivo. Nakon totalne tireoidektomije i radikalne disekcije vrata desno provedena je terapija jodom-131 koju je pacijent podnio bez većih tegoba te je preporučena nadomjesna terapija levotiroksinom. Zaključak: Metastatski papilarni karcinom u limfnim čvorovima, bez dokazanog primarnog tumora u štitnoj žlijezdi, izuzetno je rijetka dijagnoza. Ponekad se prezentira cistično promijenjenim čvorovima u području vrata koji mogu predstavljati dijagnostički problem.

Uvod: Povećanje štitnjače može biti tumorske ili netumorske prirode, a tumori mogu biti benigni i maligni. Cilj ovog istraživanja je prikazati učestalost netumorskih i tumorskih promjena štitnjače operiranih u šestogodišnjem periodu, u KBC-u Rijeka, raspodjelu po spolu, proširenosti te usporedba sa svjetskim statistikama. Materijali i metode: Iz baze podataka Zavoda za patologiju Medicinskog fakulteta Sveučilišta u Rijeci prikupljeni su podatci o tipovima patohistoloških promjena štitnjača operiranih u periodu od 2013. do 2018. godine u KBC-u Rijeka. Rezultati: U navedenom periodu operacija štitnjače izvedena je na 964 pacijenata od kojih se u 84% slučajeva radilo o ženama. Kod 46% slučajeva učinjena je totalna tireoidektomija te se u gotovo polovici slučajeva radilo o netumorskim promjenama. Od toga je u 38% slučajeva patohistološki potvrđena dijagnoza strume, a u 15% dijagnoza strume sa tireoiditisom. Benignih tumorskih promjena je ukupno bilo 30%, od čega 23% folikularna adenoma i 6% Hürthle cell adenoma. Dijagnoza malignog tumora postavljena je u 20% od ukupnog broja operiranih štitnjača. Papilarni karcinomi su bili zastupljeni sa 84% slučajeva. Prema učestalosti slijede devet slučajeva folikularnih karcinoma, devet Hürthle cell karcinoma, sedam medularnih karcinoma i dva slučaja anaplastičnog karcinoma. U jednom slučaju radilo se o SETTLE tumoru (spindle epithelial tumor with timus-like differentiation), a u dva slučaja o limfomu. Više od polovice karcinoma otkriveno je u fazi T1 (< ; 2 cm). Tumori veličine 2-4 cm (T2) otkriveni su u 30% pacijenata. 12% tumora bilo je veće od 4 cm (T3) dok je samo 3% otkriveno u fazi T4 (tumor probija čahuru i invadira okolno tkivo). Zaključak: Rezultati ove analize ukazuju na pojavu strume u polovini slučajeva, dok ostatak čine benigni i maligni tumori. Četiri puta je veća učestalost karcinoma štitnjače u ženskoj populaciji. Prevladava papilarni karcinom, najčešće otkriven u najranijoj fazi bolesti (T1). Rezultati su u skladu sa navodima svjetske literature.

INTRODUCTION: Breast cancer is one of the leading causes of cancer related deaths. In addition, immune infiltration of breast tumors has been shown to be related to clinical outcome. With respect to other infiltrating immune cells in breast cancer, a strong proportion of natural killer cells (NK) have been found in triple negative breast cancer. As part of the innate immunity, NK cells have a key role to control tumour growth by their cytotoxic activity. Based on this the goal of study was to investigate the immune profile of NK cells in order to gain a better understanding of pathological behaviour by different breast cancer subtypes. MATERIAL AND METHODS: Immunohistology was used to detect presence and localization of CD56 and IL-15 in paraffin embedded normal and tumoral breast tissue sections. The distribution and frequency of NKG2A, NKG2C, NKp46, CD94, CD69 and CD107a, was investigated in population of NK cells in mononuclear cell suspensions from peripheral blood by flow cytometry. Cytolitic mediatorꞌs mRNA was detected by quantitative RT-qPCR. RESULTS: The percentage of IL15+ and CD56+ cells were significantly higher in triple negative breast cancer tissue in comparison to luminal A and luminal B breast cancer. The frequency of NK cell activating receptors was decreased in breast cancer subtypes while inhibitory receptor (NKG2A) increased. This data correlated with decreased NK cell function, most notably cytotoxicity. Gene expression of cytolitic mediators at local level were up regulated in luminal B breast cancer. CONCLUSION: Our data show that modulation of NK cell activity at local and systemic level could be involved in pathogenesis of breast cancer. This highlights the importance of developing therapies that will be able to restore NK cell cytotoxicity to limit tumor escape from antitumor immunity. Acknowledgement: The experiments were financed by the grant No. 19-171498

Uvod: Nakon konsenzusa postignutog na konferenciji u St. Gallen-u i revizije NCCN-ovih smjernica za liječenje karcinoma dojke, u svibnju 2017. godine u Kliničkom Bolničkom Centru Rijeka, biopsija sentinel limfnog čvora uvedena je u kliničku praksu kirurškog liječenja bolesnica oboljelih od karcinoma dojke koje su neoadjuvantnim liječenjem postigle kliničku remisiju u aksili. Primarni cilj ovog istraživanja bila je evaluacija kliničkog utjecaja metode u gore navedenoj skupini pacijentica. Metode: U ovu retrospektivnu analizu uključene su sve bolesnice oboljele od karcinoma dojke, liječene u KBC Rijeka u period od svibnja 2016. do svibnja 2018., kod kojih je preoperativno provedeno sistemsko onkološko liječenje. U prvoj postoperativnoj godini praćena je pojava lokoregionalnog i sistemskog recidiva. Rezultati: Četrdeset i osam od sveukupno 65 bolesnica uključenih u analizu prezentiralo se je sa pozitivnim aksilarnim limfnim čvorovima u vrijeme postavljanja dijagnoze, od kojih je 45.83% postiglo kompletnu patološku aksilarnu remisiju. Kod ove skupine pacijentica u prvoj postoperativnoj godini nije zabilježen niti jedan slučaj lokoregionalnog recidiva, niti statistički značajna razlika u pojavnosti udaljenih metastaza u korelaciji sa opsegom kirurškog zahvata u aksili. Međutim, u komparaciji sa ovom skupinom, kod bolesnica koje nisu postigle kompletnu aksilarnu remisiju zabilježene su više stope i lokoregionalnog i distalnog recidiva. Zaključak: Biopsija sentinel limfnog čvora je pouzdana alternativa aksilarnoj disekciji za postizanje lokoregionalne kontrole i kontrole bolesti uopće, kod bolesnica oboljelih od karcinoma dojke koje neoadjuvantnim sistemskim liječenjem postignu kompletnu kliničku aksilarnu remisiju. Odgovor aksilarnih limfnih čvorova na neoadjuvantno liječenje važniji je prognostički čimbenik od statusa limfnih čvorova na početku liječenja., Introduction: After the consensus conference in St. Gallen and updated NCCN guidelines, we started doing sentinel lymph node biopsy (SLNB) in May 2017, for breast cancer patients who achieve clinical axillary remission following neoadjuvant treatment. This study’s primary goal was to evaluate the clinical impact of SLNB after neoadjuvant therapy in the group mentioned above. Methods: We retrospectively analyzed all neoadjuvant breast cancer patients from May 2016 until May 2018 at Clinical Hospital Center Rijeka. Our preliminary results recorded the appearance of locoregional and distant recurrence. Results: From 65 patients involved in this analysis, 48 patients were node-positive at the time of diagnosis, and 45.83% among those achieved complete pathological axillary remission. After the first postoperative year, there were no locoregional relapses nor statistically significant differences in the prevalence of distant recurrences, regardless of the extent of surgical procedure. However, results showed higher rates of locoregional and distant relapse for the group of patients that did not attain complete axillary remission. Conclusion: SLNB is a reliable alternative to ALND for locoregional and overall disease control for breast cancer patients who achieve complete clinical axillary remission after preoperative systemic treatment. The clinical axillary lymph node status, after neoadjuvant therapy, is a more relevant prognostic factor than the clinical axillary lymph node status at the beginning of the treatment.

Background: Tumor infiltrating lymphocytes (TILs) are involved in host imunity against tumor cells. However, in later phases of the disease high TIL infiltration is related to disease progression. Tumor immunogenicity is strongly correlated with the higher tumor mutation burden. Triple negative (TN) and HER-2 enriched breast cancers have the highest immunogenic potential so the aim of our study was to investigate the TIL infiltration and expression of PD-L1, HSP-70 in such tumors. Material and Methods: TIL infiltration was investigated in the 112 tissue samples of TN and HER-2 enriched breast cancers of women diagnosed and treated in the Clinical Hospital Centre Rijeka, Croatia, in the period between 2008 and 2016. The invasive front of the tumor (host-tumor interface), the surface layer, as well as the deep layer of the tumor were analysed. Immunohistochemistry staining of PDL-1 (SP142), HSP70 (ab2787), CD4 (SP35 Cell Marque) and CD8 (144B DakoCytomation) was performed. The results were analysed using Statistica 13 software. Results: Overall, there is a statistically significant correlation of high (over 50%) TIL infiltration with longer 5-year survival (p = 0.035, Long rank test). In the surface layer of the tumor (invasive front) there is statistically significant correlation of the intermediate TIL infiltration with the higher survival (p = 0.051, Long rank test) whereas there is no significant difference in the deep layer of the tumor. There is significant association of TIL infiltration with CD8+ T lymphocyte expression in the surface and deep layers of the tumor (Mann Whitney U test, p = 0.004 and p < 0.001, respectively), CD4+ lymphocyte expression (p < 0.001, p < 0.001, respectively) and PDL-1 expression (p < 0.001, p < 0.001, respectively). Statistically significant correlation of TIL infiltration and HSP-70 protein was only detected in the deep tumor layer (Mann Whitney U test, p < 0.001). Furthermore, in the TIL infiltrated deep tumor layer there is statistically significant positive correlation of PD-L1 and HSP-70 expression (Mann Whitney U test, p = 0.029) as well as positive European Journal of Cancer 138, Suppl. 1 (2020) correlation of the HSP-70 expression and the stage of the disease (Anova, p = 0.08). Conclusion: Although TIL infiltration in the surface layer of the tumor is corellated with higher survival rate, there is no such correlation in the deep layer. We have shown that in both layers there is increased expression of CD4 and CD8 positive T lymphocytes. However, the increased expression of inhibitory molecules PD-L1, and in the deep layer HSP-70 protein is noted as well. It is possible that in this context HSP-70 is involved in activation of Tregs and thus inducing immunotolerance to oncoproteins and along with PD-L1 molecule stimulates the development of immunosuppressive environment in the deep tumor layer thus supporting tumor immune evasion.

INTRODUCTION: Breast cancer is one of the leading cause of cancer-related deaths in women worldwide and increasing rapidly in developing countries. Heat shock proteins (HSP) and heat shock factor 1 (HSF1), key factors in the heat shock response (HSR) have been implicated in the etiology of breast cancer. Available data indicate that extracellular HSP70 has a potent motogenic activity for fibroblasts, vascular and epithelial cells, and can induce angiogenesis and matrix remodelling, suggesting a possible role in cancer development and dissemination. In the present study, we investigated the potential role of HSP70 on migration and invasion capability of breast cancer cell line in vitro. MATERIAL AND METHODS: Breast cancer cell line MDA-MB-231 was used for in vitro model study to reflect in vivo immunophenotypic features of triple negative breast cancer. Stimulation with HSP70 was tested on cancer cells lines (PANC-1, MDA-MB-231, JAR) by migration and invasion assays. The gelatinase zymography method will be used to investigate the functional activity of matrix metalloproteases 2 and 9 (MMP-2 and-9) after the invasion assay. Double immunofluorescence of actin/tubulin was investigated on HSP70 stimulated breast cancer cell lines MDA-MB-231 . RESULTS: Extracellular HSP70 induce migration and invasion in a different human cancer cell line (PANC-1, MDA-MB-231, JAR) as well as remodel of extracellular matrix by MMP-2 and-9 upregulation. CONCLUSION: These results suggest that HSP70 may facilitate cancer cell extravasation and invasion, a crucial event for carcinogenesis. Modulating cancer cell migration and invasion as well as upregulation of MMP-2 and-9, HSP70 might promote and/or sustain a permissive microenvironment for cancer cell invasion and metastasis. Acknowledgement: The experiments were financed by the grant No. 19-171498

Background: Breast cancer (BC) is the most common malignancy in women. Aim: To assess the impact of HER2 status on axillary lymph node (ALN) involvement in patients with invasive ductal carcinoma of no special type (IDC-NST) both at diagnosis and during the 4-year postoperative period. Patients and Methods: We retrospectively included 375 women with an early clinical stage of non-luminal IDC-NST who between 2007 and 2013 underwent breast surgery at a clinical hospital. They were divided into phenotype-based groups: HR+HER2-, HR+HER2+, HR-HER2+ and HR-HER2-. Only patients with sentinel lymph node (SLN) macrometastases underwent ALN dissection. If > 3 ALNs were positive, radiotherapy was delivered. All patients were treated with chemotherapy, HER2+ BC patients received trastuzumab, and hormone receptor (HR)-positive BC patients received hormonal therapy. Results: Larger tumor size, higher grade, HR+, HER2+ status, and lymphovascular invasion (LVI) were predictive for ALN metastases at diagnosis. The poorest overall, disease-free, and distant recurrence-free survival (OS, DFS, DRFS) were found in the HR-HER2- group, while the poorest locoregional recurrence-free survival (LRFS) was observed in HR-HER2+ and HR-HER2- groups. HER2 status was not predictor of survival. Conclusions: HER2+ status was predictive for ALN involvement at diagnosis but had no effect on 4-year LRFS in these patients. Antecedentes: El cáncer de mama es el tumor maligno más común en mujeres. Objetivo: Conocer el impacto del estado HER2 sobre el compromiso ganglionar axilar al momento del diagnóstico y durante los primeros cuatro años después de la cirugía en mujeres con carcinoma ductal invasivo de tipo no especial (IDC-NST). Pacientes y Métodos: Incluimos retrospectivamente a 375 mujeres en etapas clínicas iniciales de IDC-NST que fueron operadas en un hospital clínico. Ellas se dividieron en grupos de acuerdo al fenotipo: HR+HER2-, HR+HER2+, HR-HER2+y HR-HER2-. La disección de ganglios axilares se efectuó solo en las pacientes con macrometástasis en el ganglio centinela. Si había más de tres ganglios comprometidos, se efectuó radioterapia. Todas las pacientes se trataron con quimioterapia. Las pacientes HER2+ recibieron trastuzumab y las pacientes HR+ recibieron hormonoterapia. Resultados: Tumores más grandes, de mayor grado de malignidad, HR+, HER2+ y la invasión linfovascular fueron predictivos de la presencia de metástasis axilares al momento del diagnóstico. La sobrevida más baja se observó en pacientes HR-HER2+. La sobrevida libre de recurrencia locorregional más baja, se observó en pacientes HR-HER2+ y HR-HER2-. HER2 no fue predictor de sobrevida. Conclusiones: En estas mujeres, HER2+fue predictor de la presencia de compromiso ganglionar axilar al momento del diagnóstico pero no de la sobrevida a cuatro años.

Since 2016, pathologists have been collecting data on people newly diagnosed with breast cancer in Croatia. Five coordinators from large centers enter data obtained from colleagues from smaller hospitals.Here we show the data collected in 2017.

Background: Upon St. Gallen consensus conference (1) and updated version of NCCN guidelines (2) from 2017, in May 2017 in Clinical Hospital Center Rijeka sentinel lymph node biopsy (SLNB) was introduced in clinical practice for axillary staging of breast cancer patients that achieved complete clinical axillary remission after neoadjuvant systemic oncologic treatment. This trial aims to evaluate the clinical impact of SLNB performance after neoadjuvant systemic treatment (NAST) in initially node-positive breast cancer patients and to determine the prognostic value of the axillary complete pathological response. Patients and Methods: Breast cancer patients in clinical stage T1-T3 N0-N2 M0, surgically treated in our institution from September 2018 till May 2022 would be included in this trial and divided into three groups according to protocol. SLNB would be performed in all patients presenting with cN0 stage at the time of surgery, including those patients who shift from cN1-N2 to cN0 during NAST. All patients involved in this trial would be monitored for five postoperative years in order to determine following parameters: rates of local and regional recurrence, rate of disease progression to M1 stage, regional recurrence- free survival, disease progression-free survival, cancer-related mortality rate, and overall survival. Results: Results gained from this trial would be compared among groups and with our previous data of patients in equivalent stage treated in period from 2011 till 2014 when all patients were primary surgically treated, therefore axillary lymph node dissection (ALND) was performed for all node-positive patients. Conclusion: Hopefully, the results of this trial would provide enough evidence that SLNB performance after NAST does not have a negative impact on clinical outcome in breast cancer patients who had reached complete clinical axillary remission. In addition, we would try to determine the prognostic value of the axillary complete pathologic response. ClinicalTrials.gov: This protocol has been registered at clinicaltrials.gov with ID: NCT03719833 Abbreviations: SLNB=sentinel lymph node biopsy, ALND=axillary lymph node dissection, NAST=neoadjuvant systemic treatment, US=ultrasound, MRI= magnetic resonance imaging, RRR=regional recurrence rate, RRFS=regional recurrence-free survival, DFS=disease free survival, CSMR=cancer specific mortality rate, OS=overall survival

Uvod: Temeljem konsenzusa konferencije u St. Gallen-u i revidiranih NCCN-ovih smjernica za liječenje raka dojke iz 2017., u svibnju iste godine u KBC-u Rijeka biopsija sentinel limfnog čvora (SLNB) uvedena je u kliničku praksu kao metoda izbora procjene aksilarnog stadija za bolesnice oboljele od karcinoma dojke sa inicijalno zahvaćenom aksilom, a koje su neoadjuvantnim sistemskim liječenjem (NAST) postigle kompletnu kliničku aksilarnu remisiju. Kako bi evaluirali utjecaj izvođenja SLNB nakon NAST na ishod liječenja kod spomenute skupine pacijentica i pokušali objektivizirati prognostički značaj kompletnog patološkog aksilarnog odgovora (pCR) osmislili smo protokol prospektivnog opservacijskog kliničkog istraživanja. Pacijenti i metode: U istraživanje će biti uključene bolesnice oboljele od karcinoma dojke u stadiju T1-T3 N0-N2 M0, kirurški liječene u KBC Rijeka u periodu od rujna 2018. do svibnja 2022. Iste će biti podijeljene u tri grupe u skladu sa definiranim protokolom. SLNB će se učiniti kod svih bolesnica koje se preoperativno prezentiraju u stadiju cN0, uključujući i bolesnice koje su se neoadjuvantnim liječenjem konvertirale iz stadija cN1-N2 u stadij cN0. U petogodišnjem postoperativnom periodu pratit će se pojavnost lokalnog i regionalnog recidiva, progresije bolesti u M1 stadij, smrtnost od karcinoma i sveukupno preživljenje. Rezultati: Rezultati će se usporedit između skupina definiranih protokolom ovog istraživanja te sa rezultatima skupina bolesnica ekvivalentnog stadija bolesti liječenih u periodu od 2011. do 2014. kada su bolesnice bile primarno kirurški liječene, odnosno kada je svim bolesnicama sa inicijalno pozitivnim aksilarnim limfnim čvorovima standardno učinjena aksilarna disekcija (ALND). Zaključak: Dobivenim rezultatima želimo potvrditi osnovnu hipotezu ovog istraživanja; da procedura SLNB nakon NAST nema negativan utjecaj na ishod liječenja kod bolesnica sa inicijalno pozitivnom aksilom koje su preoperativnom sistemskom terapijom postigle remisiju bolesti u aksili. Uz navedeno pokušat ćemo odrediti prognostički značaj downstaging- a aksile neoadjuvantnim liječenjem. ClinicalTrial.gov: Protokol istraživanja je registriran i javno dostupan na stranicama www.clinicaltrials.gov (ID:NCT03719833), Background: Upon St. Gallen consensus conference (1) and updated version of NCCN guidelines (2) from 2017, in May 2017 in Clinical Hospital Center Rijeka sentinel lymph node biopsy (SLNB) was introduced in clinical practice for axillary staging of breast cancer patients that achieved complete clinical axillary remission after neoadjuvant systemic oncologic treatment. This trial aims to evaluate the clinical impact of SLNB performance after neoadjuvant systemic treatment (NAST) in initially node-positive breast cancer patients and to determine the prognostic value of the axillary complete pathological response. Patients and Methods: Breast cancer patients in clinical stage T1-T3 N0-N2 M0, surgically treated in our institution from September 2018 till May 2022 would be included in this trial and divided into three groups according to protocol. SLNB would be performed in all patients presenting with cN0 stage at the time of surgery, including those patients who shift from cN1-N2 to cN0 during NAST. All patients involved in this trial would be monitored for five postoperative years in order to determine following parameters: rates of local and regional recurrence, rate of disease progression to M1 stage, regional recurrence-free survival, disease progression-free survival, cancer related mortality rate, and overall survival. Results: Results gained from this trial would be compared among groups and with our previous data of patients in equivalent stage treated in period from 2011 till 2014 when all patients were primary surgically treated, therefore axillary lymph node dissection (ALND) was performed for all node-positive patients. Conclusion: Hopefully, the results of this trial would provide enough evidence that SLNB performance after NAST does not have a negative impact on clinical outcome in breast cancer patients who had reached complete clinical axillary remission. In addition, we would try to determine the prognostic value of the axillary complete pathologic response. ClinicalTrials.gov: This protocol has been registered at clinicaltrials.gov with ID: NCT03719833 Abbreviations: SLNB=sentinel lymph node biopsy, ALND=axillary lymph node dissection, NAST=neoadjuvant systemic treatment, US=ultrasound, MRI= magnetic resonance imaging, RRR=regional recurrence rate, RRFS=regional recurrence- free survival, DFS=disease free survival, CSMR=cancer specific mortality rate, OS=overall survival

Uvod: Debljina odnosno indeks tjelesne mase (ITM) je etiološki i prognostički čimbenik te ima ulogu u imunološkom odgovoru u žena oboljelih od karcinoma dojke. Cilj ove preliminarne studije je utvrditi odnos između ITM-e i odgovora na neoadjuvantno liječenje u bolesnica s karcinomom dojke te utvrditi odnos ITM-e prema imunološkom odgovoru tumor infiltrirajućih limfocita (TIL) u stromi tumora. Slučajevi i metode: U analizu je uključeno 66 bolesnica s uznapredovalim rakom dojke koje su na Klinici za onkologiju i radioterapiju KBC-a Rijeka liječene neoadjuvantnom kemoterapijom i bioterapijom te operirane od 2016. do 2017. godine. Materijal su uzorci iglenih biopsija i tkiva dojke uzetih na Klinici za radiologiju KBC-a Rijeka i na Klinici za opću i onkološku kirurgiju KBC-a Rijeka, preparati kojih su napravljeni na Zavodu za Patologiju KBC-a Rijeka, Medicinskog fakulteta Sveučilišta u Rijeci. ITM je izračunat uz pomoć formule koja koristi omjer tjelesne težine i visine bolesnica nakon čega je kategoriziran prema vrijednostima navedenim definicijom Svjetske Zdravstvene Organizacije te za potrebe statistike dodatno podijeljen u dvije skupine koje čine bolesnice normalne odnosno smanjene tjelesne težine i bolesnice povećane tjelesne težine odnosno adipozne. Na HE obojenim rezovima iglenih biopsija procijenjen je postotak stromalne limfocitne upalne infiltracije na četiri područja na granici tumora prema zdravom tkivu, te je izračunata srednja vrijednost postotka TIL-a. Nakon kompletne kirurške obrade na operacijskom materijalu je procijenjen rizik za nastanak recidiva karcinoma uz pomoć sustava Pathologic Residual Burden Calculator (PRCB) kojim se klasificira ostatni primarni tumor i ostatni metastatski tumor u lokalnom limfnom čvoru nakon neoadjuvantnog liječenja. Statistička analiza je napravljena uz pomoć programa Statistica 13.3. Rezultati: Uspoređene su dobivene vrijednosti ITM-e s PRCB odgovorom, veličinom ostatnog tumora, statusom limfnih čvorova, postotkom stromalnih limfocita, histološkim gradusom tumora u iglenoj biopsiji, imunofenotipom tumora i Ki-67 indeksom proliferacije. Nađen je pozitivan odnos ITM-e i PRCB-a (p=0.007, hi kvadrat test), ITM-e i veličine ostatnog tumora (p=0.005, T-student test) kao i ITM-e i statusa limfnih čvorova (p=0.006, hi kvadrat test). Odnos ITM-e prema histološkom gradusu tumora nije statistički značajan (p=0.211, hi kvadrat test). Dobivena je statistički značajna povezanost između ITM-e i limfocitnog upalnog infiltrata pri čemu bolesnice s većim ITM-e imaju niži postotak limfocitnog upalnog infiltrata (p=0.005, hi kvadrat test). Nađena je statistički značajna povezanost između ITM i postotka izraženosti progesteronskih receptora (PR) pri čemu porast ITM-e prati porast PR –a (p=003, Mann Whitney test). Odnos ITM i izraženosti estrogenskih receptora nije statistički značajan (p=0.085, Mann Whitney test) kao niti odnos ITM-e i Ki-67 indeksa proliferacije (p=0.481, hi kvadrat test). Zaključak: Pozitivan odnos ITM-e i izražaja hormonskih receptora prikazan je u više studija s naglaskom na poslijemenopauzalnu životnu dob. Navedeni rezultati pozitivnog međuodnosa ITM –e i veličine tumora kao i statusa limfnih čvorova odgovaraju rezultatima nekoliko studija. Međutim rezultati studija o povezanosti ITM-e i odgovora na neoadjuvantno liječenje nisu ujednačeni. Pojedine studije ITM prikazuju kao nezavisan prognostički čimbenik lošijeg odgovora na neoadjuvantno liječenje i čimbenik ranijeg povrata bolesti. Odgovor imunološkog sustava u karcinomu dojke ovisi o više čimbenika od kojih je debljina samo jedan od modulatora. Na prikazani rezultat najvjerojatnije utječe veći broj slučajeva s visokom ekspresijom hormonskih receptora u skupini visokog ITM-e koji negativno posreduju imuni odgovor.

Utjecaj količine i distribucije masnog tkiva na tumorske karakteristike invazivnog karcinoma dojke u postmenopauzalnih žena

Uvod: Mogućnost neoadjuvantnog liječenja uznapredovanog karcinoma dojke s jednakim imunofenotipom, a različitim odgovorom bolesnica na isto potiče na razmatranje utjecaja imunološkog sustava prije svega tumorskog infiltrata limfocita (TIL) u odgovoru na neoadjuvantno liječenje (1). Cilj ove preliminarne studije je bio utvrditi mogući prediktivni značaj TIL-a u odgovoru na neoadjuvantno liječenje, neovisno o molekularnim podtipovima invazivnog karcinoma dojke. Ispitanici i metode: U analizu je uključeno 30 bolesnica s uznapredovalim rakom dojke koje su na Klinici za radioterapiju i onkologiju KBC-a Rijeka liječene neoadjuvantnom kemoterapijom i bioterapijom te operirane tijekom 2016. godine. Materijal su uzorci iglenih biopsija i tkiva dojke uzetih na Klinici za radiologiju KBC-a Rijeka i na Klinici za opću i onkološku kirurgiju KBC-a Rijeka, preparati kojih su napravljeni na Zavodu za Patologiju KBC-a Rijeka, Medicinskog fakulteta Sveučilišta u Rijeci. Na HE bojenim rezovima iglenih biopsija procijenjen je postotak stromalnih TIL-a na četiri mjesta u tumoru, te je izračunata srednja vrijednost. Na operativnom materijalu procijenjen je rizik za nastanak recidiva karcinoma uz pomoć sustava Pathologic Residual Burden Calculator (PRCB) kojim se klasificira ostatni primarni tumor i ostatni metastatski tumor u lokalnom limfnom čvoru nakon neoadjuvantnog liječenja. Uspoređene su dobivene vrijednosti postotka stromalnih TIL-a s PRCB odgovorom, veličinom ostatnog tumora, statusom limfnih čvorova, histološkim gradusom tumora u iglenoj biopsiji, imunofenotipom tumora i Ki-67 indeksom proliferacije. Statistička analiza je napravljena uz pomoć programa Statistica 13.2. Rezultati: Dobivena je statistički značajna povezanost između postotka TIL-a i odgovora na neodjuvantno liječenje (p=0.04, Student T-test) pri čemu se uz patološki kompletan odgovor nalazi veći postotak limfocita. Također je nađena značajna povezanost između postotka TIL-a i veličine ostatnog tumora i statusa limfnih čvorova (p