Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Tiran, V.
Identification and characterization of putative cancer stem cells from adenocarcinoma of the lung
Doktoratsstudium der Medizinischen Wissenschaft; Humanmedizin; [ Dissertation ] 5; 2017. pp.
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- Autor*innen der Med Uni Graz:
- Betreuer*innen:
- Balic Marija
- Dandachi Nadia
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- Abstract:
- Every year, more than 1,800,000 people worldwide are diagnosed with lung cancer, and approximately 1,600,000 people die each year of this disease. Despite advances in diagnosis and treatment, lung cancer remains a fatal disease with a dismal 5-year overall survival rate of around 10%. During the past decade, technological advances have facilitated in-depth analyses of lung cancer genomes, and this knowledge has further defined the remarkable molecular and cellular heterogeneity of this disease. Specifically, intratumoral heterogeneity has been associated with therapeutic failure and drug resistance, which pose considerable clinical obstacles in finding effective treatment modalities. In this context, the cancer stem cell model has attracted a significant amount of attention in recent years as an appealing explanation for the tumor heterogeneity, drug resistance, dormancy and metastasis. The aim of the present study was to isolate, culture and characterize cancer stem cells (CSCs) in order to add new aspects to CSC biology and find new biomarkers. At the Medical University of Graz 95 patients with an adenocarcinoma of the lung were recruited. Peripheral preoperative blood and a piece of the primary tumor after resection was collected. The primary tumor was processed to isolate and enrich CSCs in cultures. Tumor cells were characterized with immunofluorescence and qRT-PCR and, subsequently, functional assays were performed. CTCs were enriched with a size-based method by filtering blood through a novel parylene microfilter system. We could establish 3 cell lines from patients with lung adenocarcinomas and 1 cell line of a squamous cell carcinoma. In the spheroid culture of each cell line we could enrich CSCs of each tumor, which could be observed as an increase of CSC markers ALDH1 and CD133, respectively. In one established cell culture, further biological evaluation was performed in a mouse model. In addition, common aberrations of lung adenocarcinomas and squamous cell carcinomas were detected with copy number variation analysis. Grown on a chorioallantoic membrane, the LT8s, LT22s and LT67s cells demonstrated invasive behavior and recapitulated the primary tumor. Our data demonstrate low efficiency of primary cell culture. However, the established primary cell cultures provide an important basis for the study of disease biology and further molecular characterization. Although these cell cultures were heterogeneous, we were able to challenge the established hypothesis and add existing knowledge on CSC biology, metastasis and resistance.