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研究生(外文):Hsiang-Yu Han
論文名稱(外文):Investigation of the disparity of chemoresistance and anti-apoptosis between colon cancer cells and colon cancer stem-like cells
外文關鍵詞:cancer stem-like cellreal-time PCRdrug resistanceanti-apoptosissphere culturecell sorting
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In recent years, the hypothesis of cancer stem-like cells (CSCs) received striking attention. This hypothesis indicated CSCs possess stem cell properties, the ability of self-renew and differentiate, and proliferate infinitely. Besides, they also have characteristics of chemotherapy-resistance and anti-apoptosis. CSCs are more difficult to eradicate and easy to cause cancer relapse because of these properties. Therefore, the mechanisms for CSCs which can escape from death after therapy merits further study.
Here, colorectal cancer cells (CRCs) were studied by investigating the characteristics of drug-resistance and anti-apoptosis within colon CSCs and non-CSCs. First, colon CSCs and non-CSCs were separated by FACS. The cytotoxicity assay showed that CSCs were more resistant to clinic chemotherapeutic drugs than non-CSCs. Moreover, the result of real-time PCR indicated that the gene expression of CSCs on certain drug resistant and anti-apoptosis was at a higher level than non-CSCs. Therefore, these findings provided practical information to design more effective therapeutic strategy for eradicating CRC.
中文摘要 Ι
英文摘要 Π
1.Introduction 1
2.Materials and methods 5
2.1 Materials 6
2.2Flow Cytometry Activated Cell Sorting ( FACSAria cell sorter) with CD133 mAb and CD44 mAb Staining 8
2.3 Sphere culture assay 9
2.4 Soft Agar Assay for Colony formation 10
2.5 Immunocytochemistry and confocal microscopy 10
2.6 Evaluation of cytotoxicity (MTT and WST-1 assay) 11
2.7 real-time PCR 12
2.8 Statistical analysis 15
3. Results 16
3.1 Flow Cytometry Activated Cell Sorting ( FACSAria cell sorter) 16
3.2 The stem cell culture condition 16
3.3 The colony formation assay 16
3.4 The proliferation ability determined by PCNA immunofluorescence staining 17
3.5 The sensitivity to chemotherapy drugs 17
3.6 real-time PCR 18
4. Discussion 19
5. Conclusion 23
6. References 24
7. Appendix 31
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