Tübingen, Universiẗat, Diss., 2006.

Thymidine kinase 1 (TK1) is a cancer biomarker which has diagnostic and prognostic potential in a variety of malignancies. TK1 is significantly elevated in the serum and tumor tissue of most malignancies. This increase in TK1 can be detected in the very early stages of malignancy, including in pre-malignant disease with an increased risk for progression. Several studies have demonstrated that elevated TK1 is found in serum months before any clinical symptoms of malignancy. It has also been demonstrated that TK1 is elevated months before clinical recurrence of malignancy. This work first sought to demonstrate the early nature of TK1 expression in breast tumor tissue and pre-malignant tissue. We found that TK1 is elevated in breast hyperplasia tissue and breast carcinoma tissue. In this study we also identified some cases of ‘normal’ tumor margins (considered normal by current pathological standards) which also had elevated TK1 expression. Conversely, true normal breast tissue from noncancerous individuals had no reported elevation in TK1 expression. This study illustrated that TK1 is elevated in pre-malignant breast hyperplasia tissue, as well as some 'normal' tumor margins. TK1 expression was significantly elevated in lung, prostate, colon, esophagus, stomach, liver, and kidney tissues. This work further investigated TK1 expression in a variety of malignant tissue including the two leading causes of cancer mortality in men: lung and prostate cancer. In our study, TK1 was significantly elevated in lung and prostate cancer but not significantly elevated in prostate hyperplasia tissue. TK1 expression also increased with increasing grade in prostate carcinoma tissue. Overall, this work demonstrated that TK1 is a good universal marker of malignancy and is elevated in early cancer development. Despite the potential for TK1 as both a screening and monitoring treatment tool, there have been significant challenges associated with developing a clinically relevant method of TK1 detection. This work proposes one clinically relevant method of detection, namely a TK1 ELISA. Using preoperable lung cancer patients and normal controls, we developed a sensitive and specific ELISA which shows highly statistically significant differences in serum TK1 levels between stage 1 and stage 2 lung cancer compared with normal controls. In fact, this TK1 ELISA is more sensitive and accurate than the traditional TK radioassay, which was unable to detect differences in TK1 between early stage lung cancer and normal patients. Although elevated TK1 is not lung cancer specific, we reported significantly elevated TK1 levels in lung cancer sputum. Screening of sputum and serum for TK1 may be one method for the early detection of lung cancer. Overall, we report TK1 has promising diagnostic potential in a variety of malignancies. We also propose one sensitive and specific method to detect TK1 levels which may easily be adapted to meet current clinical applications. We hope this work will help propel TK1 forward into clinical view in the coming years.

Understanding cancer biomarkers for diagnosis and prognosis leads to improved patient treatments and care. This thesis addresses the relevance of thymidine kinase 1 (TK1) as a cancer biomarker and the role of TK1 in cancer progression. Worldwide, cancer leads to more than 12 million deaths annually. In the United States alone, each year over 1.5 million cases will be diagnosed and over half a million persons will die. The most prevalent cancer types include skin, lung, breast, prostate, and colon. TK1 is known to be present in the serum of patients with multiple cancer types, including lung, breast, colon and prostate. In fact, it is shown to be detectable in cancer patients even before they manifest clinical symptoms. Additionally, the levels of TK1 increase progressively with increasing tumor grade; meaning that levels of TK1 can indicate tumor grade. Cellular proliferation markers such as p53 and Ki-67 have been compared to TK1 in cancer diagnosis and prognosis. TK1 has potential as both a prognostic and diagnostic biomarker in various cancer types including breast. Breast cancer is one of the most aggressive cancer types with 20-30% of diagnosed tumors becoming metastatic. Recent findings have identified additional involvement of TK1 downstream of cellular proliferation in cancer progression, including cellular invasion which is a part of cancer metastasis. These findings while efficacious, fail to identify the individual contribution of TK1 in downstream processes that aid in cancer progression. As mentioned previously, TK1 is upregulated in several different cancer types. We propose that there is an advantage to upregulated levels of TK1 in cancer progression and seek to explore its role specifically in cell invasion and survival. Based on our current understanding of TK1, we first wanted to review the history of TK1 and show the importance of understanding this crucial enzyme. Finally, we report our results from experiments exploring the influence of TK1 in vitro on breast cancer cell invasion and survival.

O saco vitelino é uma estrutura única dos vertebrados que evoluiu uma grande diversidade de características de desenvolvimento, estrutural e funcional. É considerado de fundamental importância para a sobrevivência inicial do embrião. Apesar dos intensos estudos sobre a placentação no guinea pig, ainda existem lacunas importantes para a sua compreensão, no que se refere ao saco vitelino. Esta pesquisa visa detalhar a caracterização morfológica e do desenvolvimento do saco vitelino no Cavia porcellus. Dezessete amostras foram coletadas em idades gestacionais de 12 a 55 dias e investigadas mediante histologia convencional, histoquímica, imunohistoquímica incluindo o marcador de proliferação e microscopia eletrônica de transmissão. O material histórico da implantação embrionária foi utilizado da Embryological Collection of the Museum of Natural History Berlim, Germany. Foi observada a presença temporária do saco vitelino parietal e da cavidade do saco vitelino, durante a implantação do blastocisto. O saco vitelino, que recobre a placenta corioalantoica, mostrou uma grande diferença estrutural entre o 12º e 18º dia, como a transição do epitélio pseudoestratificado para projeções, que se assemelham a uma árvore. A membrana de Reichert não está desenvolvida no 12º dia, entretanto verifica-se um espaço preenchido de material extracelular e vacúolos. A membrana de Reichert foi encontrada a partir do 16º dia de gestação e com o desenvolvimento gestacional e se tornou cada vez mais espessa e constituída de material amorfo e acelular. Os dados morfológicos sugerem que a transferência de nutrientes do saco vitelino para o feto foi mais intensa entre o 22º e 40º dia, especialmente no 40º dia. No último terço da gestação, houve uma diminuição dos tecidos de ambos os sacos vitelinos placentário e visceral. Em conclusão, o desenvolvimento do saco vitelino do guinea pig revela grande semelhança com os outros caviomorfos. Os dados sugerem que o saco vitelino nos caviomorfos é fundamental na gestação média, mas, não tanto, para os períodos iniciais da gestação, como é o caso dos outros vertebrados. / The yolk sac is a unique structure of vertebrates that evolved a great diversity of developmental, structural and functional features. It is regarded to have a fundamental role for early embryo survival. Although placentation has been intensely studied in the guinea pig, there are important gaps as in regard to the yolk sac. This research will undergo a detailed morphological and developmental characterization of the yolk sac in Cavia porcellus. Seventeen specimens have been collected at gestational ages from day 12 to 55 and investigated by conventional histology, histochemistry, immunohistochemistry including proliferation marker and transmission electron microscopy. Historical material on earlier stages was used from the Embryological Collection of the Museum of Natural History Berlin, Germany. A temporary parietal yolk sac and yolk sac cavity occurred during the blastocyst implantation. The yolk sac covering the chorioallantoic placenta showed great structural differentiation between day 12 and 18 such as a transition from a pseudostratified epithelium to projections that resemble a tree. A Reicherts membrane was not developed on day 12, but a space filled with extracellular material and vacuoles was present. The membrane was found from day 16 onward and became thick and composed of amorphous and acellular material. The morphological data suggest that a transfer of nutrients from the yolk sac to the fetus was most advanced between 22th and 40th day, especially on the 40th day. In the last third of gestation a decrease in both, placenta and visceral, yolk sacs tissues was observed. In conclusion, the development of the yolk sac in the guinea pig showed similarities to other caviomorphs. Data suggest that the yolk sac in caviomorphs is most important for mid gestation, but not so much for early pregnancy as it is for other vertebrates.

O saco vitelino é uma estrutura única dos vertebrados que evoluiu uma grande diversidade de características de desenvolvimento, estrutural e funcional. É considerado de fundamental importância para a sobrevivência inicial do embrião. Apesar dos intensos estudos sobre a placentação no guinea pig, ainda existem lacunas importantes para a sua compreensão, no que se refere ao saco vitelino. Esta pesquisa visa detalhar a caracterização morfológica e do desenvolvimento do saco vitelino no Cavia porcellus. Dezessete amostras foram coletadas em idades gestacionais de 12 a 55 dias e investigadas mediante histologia convencional, histoquímica, imunohistoquímica incluindo o marcador de proliferação e microscopia eletrônica de transmissão. O material histórico da implantação embrionária foi utilizado da Embryological Collection of the Museum of Natural History Berlim, Germany. Foi observada a presença temporária do saco vitelino parietal e da cavidade do saco vitelino, durante a implantação do blastocisto. O saco vitelino, que recobre a placenta corioalantoica, mostrou uma grande diferença estrutural entre o 12º e 18º dia, como a transição do epitélio pseudoestratificado para projeções, que se assemelham a uma árvore. A membrana de Reichert não está desenvolvida no 12º dia, entretanto verifica-se um espaço preenchido de material extracelular e vacúolos. A membrana de Reichert foi encontrada a partir do 16º dia de gestação e com o desenvolvimento gestacional e se tornou cada vez mais espessa e constituída de material amorfo e acelular. Os dados morfológicos sugerem que a transferência de nutrientes do saco vitelino para o feto foi mais intensa entre o 22º e 40º dia, especialmente no 40º dia. No último terço da gestação, houve uma diminuição dos tecidos de ambos os sacos vitelinos placentário e visceral. Em conclusão, o desenvolvimento do saco vitelino do guinea pig revela grande semelhança com os outros caviomorfos. Os dados sugerem que o saco vitelino nos caviomorfos é fundamental na gestação média, mas, não tanto, para os períodos iniciais da gestação, como é o caso dos outros vertebrados. / The yolk sac is a unique structure of vertebrates that evolved a great diversity of developmental, structural and functional features. It is regarded to have a fundamental role for early embryo survival. Although placentation has been intensely studied in the guinea pig, there are important gaps as in regard to the yolk sac. This research will undergo a detailed morphological and developmental characterization of the yolk sac in Cavia porcellus. Seventeen specimens have been collected at gestational ages from day 12 to 55 and investigated by conventional histology, histochemistry, immunohistochemistry including proliferation marker and transmission electron microscopy. Historical material on earlier stages was used from the Embryological Collection of the Museum of Natural History Berlin, Germany. A temporary parietal yolk sac and yolk sac cavity occurred during the blastocyst implantation. The yolk sac covering the chorioallantoic placenta showed great structural differentiation between day 12 and 18 such as a transition from a pseudostratified epithelium to projections that resemble a tree. A Reicherts membrane was not developed on day 12, but a space filled with extracellular material and vacuoles was present. The membrane was found from day 16 onward and became thick and composed of amorphous and acellular material. The morphological data suggest that a transfer of nutrients from the yolk sac to the fetus was most advanced between 22th and 40th day, especially on the 40th day. In the last third of gestation a decrease in both, placenta and visceral, yolk sacs tissues was observed. In conclusion, the development of the yolk sac in the guinea pig showed similarities to other caviomorphs. Data suggest that the yolk sac in caviomorphs is most important for mid gestation, but not so much for early pregnancy as it is for other vertebrates.