98 research outputs found

    Percentage of neoplastic cells in papillary thyroid carcinoma: implications for DNA microarray studies of gene expression profile

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    Badanie profilu ekspresji gen贸w technik膮 mikromacierzy DNA wykonywane jest albo w pr贸bkach guza, albo w izolowanych kom贸rkach nowotworowych uzyskanych technik膮 mikrodyssekcji. W tym przypadku uzyskuje si臋 znacznie lepsz膮 informacj臋 o ekspresji gen贸w zwi膮zanych z transformacj膮 nowotworow膮, ale zebranie wystarczaj膮cej ilo艣ci nieuszkodzonego RNA jest bardzo trudne i w praktyce klinicznej niemo偶liwe. Wyj艣ciem kompromisowym jest pr贸ba odniesienia profilu ekspresji gen贸w do procentowego udzia艂u kom贸rek nowotworowych w utkaniu guza. Rak brodawkowaty tarczycy (PTC, papillary thyroid carcinoma) nale偶y do tych guz贸w nowotworowych, kt贸re charakteryzuj膮 si臋 du偶ym udzia艂em pod艣cieliska. Udzia艂 kom贸rek nowotworowych w utkaniu raka brodawkowatego oceniano przez przybli偶one badanie liczby kom贸rek widocznych w 10-18 polach widzenia oraz badanie korelacji mi臋dzy t膮 informacj膮 a profilem ekspresji gen贸w, uzyskanych metod膮 mikromacierzy DNA. Badanie wykonano w 40 przypadkach raka brodawkowatego tarczycy i stwierdzono, 偶e odsetek kom贸rek PTC waha si臋 od 20-95% i tylko w oko艂o 1/4 przypadk贸w przekracza 75%. Wykazano, 偶e trafne rozr贸偶nienie profilu ekspresji guza nowotworowego od prawid艂owego utkania tarczycy jest mo偶liwe, gdy odsetek kom贸rek nowotworowych przekracza 25-30%.Sygna艂 informacyjny wynikaj膮cy z badania profilu ekspresji gen贸w metod膮 mikromacierzy DNA jest w raku brodawkowatym tarczycy bardzo silny i pozwala na prawid艂owe rozr贸偶nienie utkania nowotworowego od utkania prawid艂owego nawet w przypadkach, gdy odsetek kom贸rek nowotworowych waha si臋 w zakresie 25-50%. Wniosek ten zach臋ca do kontynuacji bada艅 mikromacierzowych, gdy偶 silnie wskazuje na mo偶liwo艣膰 uzyskania sygnatury genowej przydatnej w rutynowej diagnostyce raka brodawkowatego tarczycy w materiale uzyskanym z biopsji podejrzanego guza.Studies of gene expression profile using DNA microarray technology are usually performed using either tumor-derived sample material or isolated neoplastic cells obtained through microdissection. The scope of information about neoplastic transformation gained from studying profile of gene expression in microdissected samples would be much wider but collection of sufficient amounts of intact RNA is very difficult. A compromise could be reached by relating gene expression profile to percentage of neoplastic cells in the investigated tissue sample. The ratio of neoplastic cells in the investigated sample of papillary thyroid cancer was assessed through evaluation of approximated count of cell number in 10-18 examined image fields. This information was related to gene expression profiles obtained from DNA microarrays. The study involved 40 cases of papillary thyroid cancer; the percentage of PTC cells varied between 20 and 95% and only in half of the cases exceeded 75%. Correct differentiation of tumor and normal sample by means of gene expression profile was possible only when the percentage of tumor cells exceeded 25-30%. Seventeen genes showing the best correlation with the tumor cell numbers were selected and their classification potential was evaluated.Strength of information derived from gene expression profile studies by DNA microarrays in papillary thyroid cancer cells is very reliable and permits distinguishing correctly between normal and neoplastic tissues even when the percentage of cancer cells does not exceed 25-50%. However, the differentiation potential of gene expression profile is not markedly improved by selection of genes showing best correlation with the number of tumor cells

    Chemotherapy in disseminated thymoma - case report, own experience

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    Grasiczaki nale偶膮 do nowotwor贸w, kt贸re powstaj膮 z tkanki nab艂onkowej grasicy. Zabieg chirurgiczny odgrywa g艂贸wn膮 rol臋 w leczeniu grasiczak贸w, ale chemioterapia mo偶e znacz膮co wp艂ywa膰 na polepszenie prognoz dla tej grupy pacjent贸w. Skuteczna chemioterapia mo偶e mie膰 charakter leczenia neoadjuwantowego przed zabiegiem operacyjnym, uzupe艂niaj膮cego po zabiegu, stanowi膰 element leczenia skojarzonego z radioterapi膮 lub by膰 stosowana w przebiegu rozsiewu choroby. Celem pracy jest przedstawienie przypadku 66-letniego chorego, u kt贸rego rozpoznano zaawansowanego grasiczaka i zastosowano z dobrym efektem leczenie systemowe dw贸ch linii. Pneumonol. Alergol. Pol. 2011; 79, 3: 222-226Thymomas are thymic epithelial neoplasms. Surgery plays a major role in thymoma treatment but chemotherapy can significantly improve prognoses for this group of patients. Neoadjuvant chemotherapy (before surgery), adjuvant chemotherapy (after surgery), chemotherapy combined with radiotherapy and palliative chemotherapy in dissemination stage could be required in effective therapy. The aim of this paper is description of the effective systemic second line treatment in the case of 66 years old patient with advanced thymoma. Pneumonol. Alergol. Pol. 2011; 79, 3: 222-22

    Germinal mutations of RET, SDHB, SDHD, and VHL genes in patients with apparently sporadic pheochromocytomas and paragangliomas

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    Introduction: Pheochromocytomas and paragangliomas are derived from neural crest cells and are localized mainly in adrenal medulla and sympathetic or parasympathetic ganglia. They can be inherited (25%) and be part of multi-endocrine syndromes such as MEN2 syndrome, von Hippel-Lindau syndrome, pheochromocytoma/paraganglioma syndrome, neurofibromatosis type 1, and Sturge-Weber syndrome. Clinical presentation can sometimes be atypical and does not always allow proper diagnosis. In such situations, DNA analysis can be helpful, especially when the pheochromocytoma is the first and only symptom. Material and methods: We analyzed DNA from 60 patients diagnosed and treated in the Centre of Oncology with a diagnosis of pheochromocytoma or paraganglioma. DNA analysis was carried out for RET (exons 10, 11, 13, and 16), SDHB, SDHD, and VHL genes. Techniques used for the analysis were direct sequence analysis, MSSCP, and RFLP. Results: Germinal mutations were found in 16 patients (26,7%). Most frequent were mutations in RET proto-oncogene, followed by VHL gene, one mutation in SDHB, and one in SDHD genes. A comparison of some of the clinical features of both groups (with and without mutation) showed statistically significant differences. Conclusions: The results of our study show that genetic predisposition is frequent in chromaffin tissue tumours, which indicates that DNA analysis is necessary in every case, also because of possible atypical clinical presentation. (Pol J Endocrinol 2010; 61 (1): 43-48)Wst臋p: Guzy chromoch艂onne i nerwiaki przyzwojowe wywodz膮 si臋 z kom贸rek grzebienia nerwowego i zlokalizowane s膮 g艂贸wnie w rdzeniu nadnerczy oraz w zwojach autonomicznych wsp贸艂czulnych i przywsp贸艂czulnych. Mog膮 one (25%) stanowi膰 sk艂adnik zespo艂贸w wielogruczo艂owych, takich jak zesp贸艂 MEN2, zesp贸艂 von Hippela-Lindaua, zesp贸艂 mnogich guz贸w chromoch艂onnych i nerwiak贸w przyzwojowych (PPS, pheochromocytoma/paraganglinoma syndrome), a tak偶e nerwiakow艂贸kniakowato艣ci typu 1 czy zespo艂u Sturge-Webera. Nie zawsze obraz kliniczny pozwala jednoznacznie okre艣li膰 rodzaj zespo艂u, dlatego badanie DNA mo偶e by膰 pomocne w ustaleniu rozpoznania. Celem pracy by艂a ocena cz臋sto艣ci wyst臋powania dziedzicznie uwarunkowanych guz贸w chromoch艂onnych i przyzwojak贸w u zg艂aszanych jako pierwszy objaw. Materia艂 i metody: Przeanalizowano DNA pochodz膮ce od 60 chorych leczonych i diagnozowanych w Centrum Onkologii z powodu guza chromoch艂onnego i nerwiak贸w przyzwojowych. Przeprowadzono analiz臋 DNA w zakresie nast臋puj膮cych gen贸w: RET (eksony 10, 11, 13 i 16), SDHB, SDHD i VHL. Wykorzystywano sekwencjonowanie DNA, analiz臋 MSSCP oraz analiz臋 restrykcyjn膮. Wyniki: Mutacje germinalne znaleziono u 16 chorych (26,7%). Najcz臋stsze by艂y mutacje w genie RET, nast臋pnie w genie VHL oraz po jednej mutacji w genach SDHB i SDHD. Analiza danych klinicznych chorych b臋d膮cych nosicielami mutacji wykaza艂a znamienne statystycznie r贸偶nice w por贸wnaniu z grup膮 chorych z guzami sporadycznymi. Wnioski: Przeprowadzone badania wskazuj膮 na cz臋sty udzia艂 predyspozycji dziedzicznej w wyst膮pieniu guz贸w chromoch艂onnych, co wskazuje na konieczno艣膰 wykonywania badania DNA w ka偶dym przypadku, tak偶e ze wzgl臋du na mo偶liwo艣膰 nietypowego przebiegu klinicznego, zw艂aszcza w grupie chorych w wieku 20–40 lat. (Endokrynol Pol 2010; 61 (1): 43-48

    Cz臋sto艣膰 wyst臋powania mutacji somatycznych RAS w raku rdzeniastym tarczycy 鈥 analiza populacji polskiej

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    Introduction: Somatic RET mutations are detectable in two-thirds of sporadic cases of medullary thyroid cancer (MTC). Recent studies reported a high proportion of RAS somatic mutations in RET negative tumours, which may indicate RAS mutation as a possible alternative genetic event in sporadic MTC tumorigenesis. Thus, the aim of the study was to evaluate the frequency of somatic RAS mutations in sporadic medullary thyroid cancer in the Polish population and to relate the obtained data to the presence of somatic RET mutations.Material and methods: Somatic mutations (RET, RAS genes) were evaluated in 78 snap-frozen MTC samples (57 sporadic and 21 hereditary) by direct sequencing. Next, three randomly selected RET-negative MTC samples were analysed by the next generation sequencing.Results: RAS mutation was detected in 26.5% of 49 sporadic MTC tumours. None of all the analysed samples showed N-RAS mutation. When only RET-negative samples were considered, the prevalence of RAS mutation was 68.7%, compared to 6% observed in RET-positive samples. Most of these mutations were located in H-RAS codon 61 (72%). None of 21 hereditary MTC samples showed any RAS mutations.Conclusions: RAS mutations constitute a frequent molecular event in RET-negative sporadic medullary thyroid carcinoma in Polish patients. However, their role in MTC tumorigenesis remains unclear. (Endokrynol Pol 2015; 66 (2): 121鈥125)Wst臋p: Somatyczne mutacje proto-onkogenu RET wykrywane s膮 w trzech czwartych wszystkich sporadycznych rak贸w rdzeniastych tarczycy (MTC). Ostatnie badania wykaza艂y, 偶e mutacja genu RAS jest r贸wnie偶 cz臋stym wydarzeniem w sporadycznych guzach MTC, co mo偶e oznacza膰, 偶e mutacje gen贸w z rodziny RAS s膮 alternatywnym wydarzeniem molekularnym w kancerogezie sporadycznej postaci tego raka. Z tego wzgl臋du celem niniejszej pracy by艂o oszacowanie cz臋sto艣ci wyst臋powania mutacji gen贸w RAS w sporadycznym raku rdzeniastym tarczycy w populacji polskiej i odniesieniu cz臋sto艣ci ich wyst臋powania do obecno艣ci mutacji somatycznych proto-onkogenu RET.Materia艂 i metody: Materia艂 do bada艅 stanowi艂o 78 fragment贸w guza raka rdzeniastego tarczycy (57 pr贸bek postaci sporadycznej i 21 dziedzicznej MTC). Analizowano mutacje genu RET, H-RAS, K-RAS i N-RAS metod膮 bezpo艣redniego sekwencjonowania a tak偶e 3 pr贸bki raka sporadycznego, wybrane losowo, zosta艂y zeskwencjonowane metod膮 g艂臋bokiego sekwencjonowania (Illumina).Wyniki: Mutacj臋 gen贸w RAS wykryto w 26,5% z 49 przeanalizowanych guz贸w sporadycznej postaci MTC. Natomiast, gdy tylko brano pod uwag臋 pr贸bki RET-negatywne, cz臋sto艣膰 wyst臋powania mutacji gen贸w RAS wynosi艂a 68,7% w por贸wnaniu z 6% obserwowanych w guzach RET-pozytywnych. Nie wykryto, w 偶adnej z pr贸bek, mutacji genu N-RAS. Najcz臋艣ciej wykrywan膮 mutacj膮 by艂a zmiana w kodonie 61 genu H-RAS (72%). Nie wykryto mutacji gen贸w RAS w 偶adnej z pr贸bek dziedzicznego guza raka tarczycy.Wnioski: Mutacje somatyczne gen贸w RAS s膮 cz臋stym wydarzeniem obserwowanym w RET-negatywnych sporadycznych rakach rdzeniastych tarczycy w populacji polskiej. Jednak偶e rola tych mutacji w rozwoju rdzeniastego raka tarczycy nie jest do ko艅ca poznana. (Endokrynol Pol 2015; 66 (2): 121鈥125

    Ratio of proliferation markers and HSP90 gene expression as a predictor of pathological complete response in breast cancer neoadjuvant chemotherapy

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    Introduction. Prediction of response to preoperative breast cancer chemotherapy may offer a substantial optimization of medical management of this disease. The most efficient prediction would be done a priori, before the start of chemotherapy and based on the biological features of patient and tumor. Numerous markers have been proposed but none of them has been applied as a routine. The role of MKI67 and HSP90 expression has been recently suggested to predict treatment sensitivity in HER2-positive breast cancer. The aim of this study was to validate the utility of proliferation based markers (MKI67 and CDK1) and heat shock proteins (namely HSP90) to predict response to chemotherapy in cohort of breast cancer patients treated preoperatively. Material and methods. Ninety-three patients with breast cancer, all females, mean age 42.2 years, among them 32% T1-T2 patients, 49% T3 patients and 13% with T4 tumor stage, 27% N0, 42% N1, 16% N2, 15% N3 were subjected to initial chemotherapy. The majority of patients (86%) received anthracycline and taxane chemotherapy. Among the patients there were 9 individuals with metastatic disease (M1) at initial presentation, and 11 patients were not treated surgically after initial chemotherapy (no sufficient disease response). From 82 patients operated on, 20 patients (24%) showed pathological complete response (pCR), while in 62 patients there was no pCR. 42% of patients were hormone-sensitive HER2-negative, 20% hormone-sensitive HER2-positive, 9% only HER-positive and 29% with triple negative breast cancer. Four gene transcripts (MKI67, cyclin-dependent kinase 1 [CDK1], heat shock proteins HSP90AA1 and HSP- 90AB1) were analyzed in total RNA isolated from single core obtained during preoperative core needle biopsy by quantitative real-time PCR with fluorescent probes (Universal Probe Library, Roche). Results were normalized to the panel of reference genes. Results. There were no statistically significant differences in MKI67 and CDK1 expression between pCR and no pCR groups (p = 0.099 and 0.35, respectively), although the median expression of both genes was slightly higher in pCR group. In contrast, both HSP90AA1 and HSP90AB1 transcripts showed decreased expression in pCR group (medians 0.77 and 0.55) when compared to no p CR group (median 0.86 and 0.73), statistically significant for HSP90AA1 (p = 0.031) and of borderline significance for HSP90AB1 (p = 0.054). The most significant predictor of pCR was the ratio of CDK1 transcript to HSP90AA transcript. This ratio was significantly higher in CR group (median 0.99) than in no CR group (median 0.68, p = 0.0023), and showed a potential diagnostic utility (area under receiver operating characteristic [ROC] curve 0.72). Conclusions. HSP90AA1 and AB1 genes exhibit low expression in breast cancers highly sensitive to chemotherapy and may indicate the patients with higher probability of pathological complete response. The ratio of HSP90AA1 to proliferation-related markers (CDK1 or MKI67) may be even better predictor of pCR chance, with higher expression of proliferation genes and lower stress response in patients sensitive to chemotherapy

    Myxoma virus expressing LIGHT (TNFSF14) pre-loaded into adipose-derived mesenchymal stem cells is effective treatment for murine pancreatic adenocarcinoma

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    Pancreatic ductal adenocarcinoma (PDAC) is a weakly immunogenic fatal neoplasm. Oncolytic viruses with dual anti-cancer properties鈥攐ncolytic and immune response-boosting effects鈥攈ave great potential for PDAC management. Adipose-derived stem cells (ADSCs) of mesenchymal origin were infected ex vivo with recombinant myxoma virus (MYXV), which encodes murine LIGHT, also called tumor necrosis factor ligand superfamily member 14 (TNFSF14). The viability and proliferation of ADSCs were not remarkably decreased (1鈥2 days) following MYXV infection, in sharp contrast to cells of pancreatic carcinoma lines studied, which were rapidly killed by the infection. Comparison of the intraperitoneal (IP) vs. the intravenous (IV) route of ADSC/MYXV administration revealed more pancreas-targeted distribution of the virus when ADSCs were delivered IP to mice bearing orthotopically injected PDAC. The biodistribution, tumor burden reduction and anti-tumor adaptive immune response were examined. Bioluminescence data, used to assess the presence of the luciferase-tagged virus after IP injection, indicated enhanced trafficking into the pancreata of mice bearing orthotopically-induced PDAC, as compared to tumor-free animals, resulting in extended survival of the treated PDAC-seeded animals and in the boosted expression of key adaptive immune response markers. We conclude that ADSCs pre-loaded with transgene-armed MYXV and administered IP allow for the effective ferrying of the oncolytic virus to sites of PDAC and mediate improved tumor regression
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