| Summary: | 碩士 === 國立陽明大學 === 生物醫學影像暨放射科學系暨研究所 === 99 === Tumors are organized in a heterogeneous manner with a subpopulation of limited number(less than 5%) called Cancer stem cells (CSCs) with the ability to self-renewal and giving rise to a differentiated progeny that represents most of the tumor populations. CSCs are chemoresistant and metastatic, two features that very likely contribute to the poor prognosis of advanced cancer. Furthermore, CSCs are also the key player in tumor recurrence. Therefore, exploring cell markers or function specific for CSCs identification is urgently needed.
Recently, Leung EL et al have showed that CD44 is a tumor stem cell-associated marker in non-small cell lung cancer (NSCLC) H1299 cell, but their data showed up to 81.3% of CD44+ H1299 cells with cancer stem cell characteristics. However, this huge population of cells is quite different with the definition of CSCs. Thus, in this study, we start to seek CSCs of H1299 by other methodology. Aldehyde dehydrogenase class 1A1 (ALDH1A1) has been identified as highly expressed in normal and cancer stem cells, although the population of ALDH+ is hardly be detected in H1299 cells. Based on the fact that CSCs can be enriched by spheroid culture technique, we thus employ the spheroid culture system firstly and then using the ALDEFLUOR assay to mark or identify the CSCs from H1299 cells. To label and identify cells in vivo and in vitro, we also established two cell lines that stably expressed a dual (DsRedm-tTKsr39) or a triple fused reporter construct (mcherry-Fluc-tTKsr39), which is driven from the STAT3 and NF-κB hybrid promoter and Oct4 promoter, respectively.
Overall, using these reporter systems will allow us for tracking CSCs in the real-time manner so that improve our understanding on the molecular mechanism of CSCs and lead to progress in cancer management.
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