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The Establishment of Vincristine-resistant Human Sarcoma Cell Line (MS5/VCR)

Author: WangPengPeng
Tutor: YangLiYe
School: Shantou University
Course: Pediatrics
Keywords: Cancer stem cells Multidrug resistance Vincristine Sarcoma cell lines Abnormal hemoglobin Hb G - Chinese PCR Restriction endonuclease
CLC: R73-3
Type: Master's thesis
Year: 2009
Downloads: 15
Quote: 0
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Abstract


Objective: To establish in vitro human sarcoma the MS5 cell lines vincristine resistant cell line , observe the biological characteristics and in vivo tumorigenicity . Methods: In this thesis, the low concentration plus continued induce the established sarcoma MS5 cells resistant to vincristine model , named MS5/VCR . Light microscope morphology of the cells , in vitro drug sensitivity test two cells sensitive to chemotherapeutic drugs , MS5 and MS5/VCR of cells subcutaneously inoculated nude mice , kill the tumor in nude mice 55 days after the formation of the size of the tumor , immunohistochemistry was used to observe the transplanted tumor MDR - 1 expression . MS5 by the VCR of the results : sarcoma cells induced after seven months to obtain vincristine - resistant cell lines Sarcoma MS5/VCR MS5/VCR 6μg/ml medium vincristine concentration of stable growth , compared with MS5 cells resistant cell line slightly rounded shape . In vitro drug sensitivity test results on the VCR Show MS5/VCR cell lines have a certain resistance to tumor seeding experiments display MS5/VCR cells than MS5 cells tumorigenicity strong immunohistochemical results mdr-1 MS5 and MS5 / VCR transplanted tumors were expressed . Conclusion: 1 , in vitro concentration gradient incremental approach can be induced to establish sarcoma the MS5 vincristine resistant cell line MS5/VCR . 2, sarcoma the MS5 vincristine resistant cell line MS5/VCR clone in vitro and in vivo tumorigenicity ability to stronger than parental cells MS5 may contain more cancer stem cells . Purpose : the application of the experimental hematology and DNA analysis identified three cases of abnormal hemoglobin Hb G - Chinese . Methods: cellulose acetate membrane electrophoresis method found abnormal hemoglobin . Polymerase chain reaction , restriction enzyme digestion and DNA sequence determination to determine the presence of HB G - Chinese gene mutation . Results: The three cases HB G - Chinese carriers . Conclusion : PCR to determine gene mutations leading to hemoglobin variants a simple , fast, and inexpensive method , HB G - Chinese carriers of α-globin-2 G to C alternative to a pst I restriction sites .

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