Browsing by discipline "Onkologi"

Radiation induced tumor cell death is strongly dependent on oxygen. As abnormal tumor vasculature promotes tumor hypoxia, drugs that induce vascular normalization, such as the anti-vascular endothelial growth factor (VEGF) antibody, have been tested as radiation sensitizers in preclinical and clinical settings. The insufficient benefit obtained with anti-VEGF therapy prompted us to test if antibodies blocking the endothelial growth factor angiopoietin-2 (Ang2) could improve the effect of radiation in mouse tumor allografts and human tumor xenografts. Mouse or human tumor cells were injected subcutaneously in isogenic immunocompetent or immunodeficient (NSG) mice, respectively, and tumors were allowed to form. The mice were injected with anti-Ang2 or control antibodies every three or four days starting three days before 3x2 Gy or 4x0.5 Gy whole-body radiation, followed by analysis of tumor growth, histology, vasculature, hypoxia and necrosis. Combination treatment with anti-Ang2 and radiation improved tumor growth inhibition and extended the survival of mice with melanoma or colorectal carcinoma allografts. A similar anti-Ang2 plus radiation response was also obtained in immunodeficient mice implanted with a human colorectal carcinoma xenograft, indicating that the adaptive immune response was not essential for the effect. Histological and immunohistochemical analysis of the tumors showed that the combination treatment decreased tumor vasculature, and increased tumor hypoxia and tumor necrosis in comparison with control tumors and tumors treated with the monotherapies. Our results suggest that a combination of Ang2 blocking antibodies and radiation increases tumor growth inhibition and extends the survival of tumor-bearing mice. Significance: These findings offer a preclinical rationale for further testing of the use of Ang2 blocking antibodies in combination with radiation to improve the overall outcome of cancer treatment.

Background: 5-year survival rate of oral tongue squamous cell carcinoma (OTSCC) has been low (less than 60%) despite developing treatment modalities. A previous research revealed that different populations of inflammatory cells infiltration in OTSCC were associated with different clinical outcomes. On the other hand, extracellular vesicles (EVs) secreted by OTSCC cells suggested crosstalk between OTSCC cells and tumor infiltrating inflammatory cells. Study aims: This study aims to investigate the interaction between OTSCC cells and inflammatory cells and answer 3 questions: (1) Can human peripheral blood mononuclear cells (MNCs) affect activities of OTSCC cells such as proliferation, migration and invasion? (2) Can EVs of OTSCC cells affect polarization of macrophages? (3) Can EVs of OTSCC cells affect cytotoxic activity of CD8+ T cells and NK cells? Materials and methods: Two OTSCC cell lines (HSC-3 and SCC-25) were used. OTSCC cells and human peripheral blood MNCs were co-cultured using a 3D organotypic myoma model. Proliferation and invasion into myoma tissue of OTSCC cells were detected by Immunohistochemical staining of pan-cytokeratin and Ki67. Invasion area and depth of OTSCC cells were measured using ImageJ software. Migration of OTSCC cells in the presence of MNCs was monitored using a scratch wound healing assay with IncuCyte™ system. OTSCC EVs were isolated with ultracentrifugation and characterized with NTA and Immuno-EM. Human primary monocytes, CD8+ T cells and NK cells were isolated using MACS, and their purity was checked using FACS. Expression of macrophage phenotypic markers was checked with qPCR. Cytotoxic activity was evaluated using an IncyCyte™ cell killing assay. Results: Activated human peripheral blood MNCs significantly reduced proliferation of both OTSCC cell lines, and invasion area of only HSC-3. None of the inflammatory cells in the experiment had any effect on invasion depth and migration of OTSCC cells. On the other hand, OTSCC cell-derived EVs didn't influence macrophage polarization, but had heterogeneous modulating effects on cytotoxic activity of CD8+ T cells and NK cells. Conclusion: We detected effects of OTSCC cells and inflammatory cells on each other by secreted molecule mediators or EVs, but the results were not uniform and varied in different OTSCC cell lines or inflammatory cell populations and sources. The outcome of the study emphasizes the importance of a personalized design of cancer treatment, which takes other components in tumor microenvironment such as inflammatory cells and EVs into consideration.

Kolesteroli on solukalvojen välttämätön rakennemolekyyli ja sen pitoisuus solukalvolla vaikuttaa muun muassa kasvutekijäsignalointiin. MLN64 (STARD3) on proteiini, joka kykenee siirtämään kolesterolia solun eri kalvojen välillä ja jonka yli-ilmentyminen soluissa muuttaa solukalvon kolesterolipitoisuutta ja voi siten vaikuttaa solukalvon signaalireitteihin. HER2 (erbB-2) on tyrosiinikinaasireseptoreihin kuuluva HER-ryhmän solukalvoreseptori. HER2-positiivisuus rintasyövässä on syövän ennustetta huonontava tekijä. HER2-positiiviset rintasyövät ilmentävät lähes poikkeuksetta voimakkaasti myös MLN64:ä. Tutkielmani tarkoituksena oli selvittää, vaikuttaako rintasyövän MLN64-proteiinin ilmentyminen kolesterolisynteesin avainentsyymin, HMG-CoA-reduktaasin (HMGCR), tai steroidisynteesin pullonkaulaentsyymin, CYP11A1:n, lähetti-RNA-määriin (mRNA-määriin). CYP11A1 ei fysiologisena ilmenny rintarauhaskudoksessa ja sen ilmeneminen voisi mahdollistaa rintasyövän oman steroidisynteesin. Materiaalina oli FinHer-aineiston formaliinifiksoiduista ja parafiiniin valetuista rintasyöpänäytteistä eristetty RNA. RNA käännettiin cDNA:ksi kvantitatiivista PCR-reaktiota varten, jolla tutkittiin näytteiden HMGCR:n ja CYP11A1:n lähetti-RNA-tasot. Tutkielman mukaan MLN64:n voimakas ilmentyminen korreloi positiivisesti HMGCR:n mRNA-määrään. CYP11A1 ilmentyi 16 % näytteistä, mutta korrelaatiota MLN64:n määrään ei havaittu. Tutkimus tukee näkemystä, jonka mukaan voimakas MLN64:n ilmentyminen vaikuttaa syöpäsolujen kolesterolituotantoon.

Miesten rintasyöpä on hyvin harvinainen sairaus, ja sen syyt tunnetaan huonosti. Perinnöllisillä tekijöillä saattaa olla miesten rintasyövän synnyssä tärkeä rooli, ja perheen syöpähistoria on havaittu epidemiologisissa tutkimuksissa merkittäväksi riskitekijäksi. Perinnölliset mutaatiot BRCA1- ja etenkin BRCA2-geeneissä aiheuttavat myös miehille kohonneen riskin. Myös muita, pienemmän riskin aiheuttavia alleeleja on tunnistettu viime vuosina. Niiden kliininen merkitys eri populaatioissa vaihtelee suuresti. Tässä tutkimuksessa genotyypitettiin 72 miespuolisen rintasyöpäpotilaan sekä yhdeksän perheen, joissa esiintyy miesten rintasyöpää, perimä kiinnostuksen kohteina olleiden mutaatioiden suhteen CHEK2-, PALB2-, RAD51C-, RAD51D- ja FANCM-geeneissä.

Next-generation sequencing has evolved during the past 10 years to become the go-to method for genome-wide analysis projects. Based on parallelizable PCR methods adopted from the traditional Sanger sequencing, NGS platforms can produce massive amounts of genetic information in a single run and read an entire DNA molecule within a day. The immense amount of nucleotide sequence data produced by a single sample has brought us to an era of algorithmic optimization for analysis and guring out parallelization schema. For cohort projects generally cloud based systems are used due to vast computing power requirements. Anduril is an integration and parallelization framework well suited for NGS analysis, as is shown in this study. After a brief review of the golden standard methods of NGS analysis, we describe the incorporation of the main tools into the new sequencing bundle for Anduril. Tools for alignment (BWA, Bowtie), recalibration (GATK, Picard-tools) and variant calling (GATK, Samtools, VarScan) are in main focus. The Best Practice of Broad Institute, creators of The Genome Analysis Toolkit (GATK), has been a big inspiration in the creation of our sequencing pipeline. The evolution of sequencing bundle tools into a pipeline is discussed through three separate project examples. First, a small group of 8 chronic myeloid leukemia patient samples were analysed after implementation of the main tools of the pipeline. The results were consistent with previous results, but no novel relevant mutations were found. Second, exome sequencing data from 180 breast cancers with controls available in TCGA (The Cancer Genome Atlas) were processed for use in various projects in our lab. The example showed the power of Anduril in gross cohort analysis projects, enabling automatic parallelization and intelligent work ow management system. Third, we analysed exome data from 330 TCGA ovarian cancers with controls and created a prototypical set of database components for creation of a database of annotated variants for use in analytical queries. Compared to other integration frameworks (e.g. GATK, Crossbow and Hadoop), Anduril is a robust contender for the programming oriented scientist. As cloud computing is becoming at an increasing rate a requirement in large genome-wide analysis projects, Anduril provides an e ective generalizable framework for adding tools, creating pipelines and executing entire work ows on multi-nodal computing servers. As technology advances and available computational resources grow, fast multi-processor analysis can be incorporated into health care more and more for detection of disease causing genes, medication kinetics altering polymorphisms and cancer driving mutations in an everyday setting.

Hudmelanom är en malign sjukdom som afficierar huden. Tumören infiltrerar kringliggande vävnader och sprids genom metastasering, främst via lymfkärl till lymfknutor. Vid utredning av melanom är det viktigt att utreda om sjukdomen spridit sig till de närmaste lymfknutorna, portvaktskörtlarna. Ifall metastaser konstateras i portvaktskörtel utförs körtelutrymning av det lymfknutområde där metastaser konstaterats. Målet med studien är att granska metastasfynd i portvaktskörtel från åren 2012 och 2013 för att utreda hur metastasfyndet påverkar patientens prognos samt hur ofta ytterligare metastaser konstateras i körtelutrymning. I studien konstaterades metastaser i portvaktskörtel hos 16,7 % av patienterna. Hos 16,7 % av patienterna konstaterades metastaser i körtelutrymning. Överlevnadsprognosen för patienter med metastaser i portvaktskörtel motsvarade resultat i tidigare studier. Övriga faktorer som försämrade prognosen bland patientmaterialet var hög ålder, stor metastas och lokalrecidiv. Studiens material bestod av 42 patienter, för att kunna fatta nya vårdbeslut om melanom krävs studier med större patientmaterial.

The majority of cancers, such as breast cancer, originate from epithelial structures. Highly organized epithelial tissues are comprised of cells which manifest apico-basal polarization. Factors regulating apico-basal polarity and epithelial integrity are often observed deregulated in cancer cells and loss of polarity is often observed in tumors. However, the importance and the specific role of epithelial integrity regulators in tumorigenesis are still not fully defined. This study shows that the key regulators of epithelial cell polarity Par6B and Par6G proteins have a role in the restriction of cell proliferation in mammary epithelial cell lines. Using the shRNA silencing approach, downregulation of PARD6B and PARD6G in the cells lead to the impediment of the cell-cycle exit, verified in proliferation suppressive conditions in which cells normally would enter quiescence. Par6 proteins were shown to regulate cell proliferation via the canonical PI3K/Akt pathway, which is one of the most commonly deregulated cell proliferation promoting pathways in cancer cells. The results demonstrate the unknown function of Par6B and Par6G proteins as cell proliferation regulators and a previously unrecognized relation between Par6 proteins and PI3K/Akt pathway. However, the detailed interaction between Par6 proteins and the PI3K/Akt pathway ought to be investigated further. In addition, the results revealed that Par6 proteins have different effects on cell proliferation suggesting biological differences between Par6 proteins and that certainly bears further investigation. In conclusion, the study presents a previously unknown connection between epithelial integrity regulators and cancer-relevant cell proliferation promoting pathways, which may provide new targets for therapeutic intervention in the future.