| Multidrug resistance (MDR) is a phenomenon that tumor cell can produce resistance to many kind of chemotherapeutic drugs, include these construction, fuction and kill mechanism are different chemotherapeutic drugs. the production of MDR1 of tumor tissue result from MDR1 gene unnormal expansion and over expression. It's expression product is glycoprotein and molecular weight is 170KB, so it is named P-glycoprotein (P-gp). P-gp is a exhaust pump depend on ATP that can exhaust large molecular medicine and toxicant in cells. It make tumor cells resistance to chemotherapeutic drugs. we utilize this character of MDR1, adopt adenovirus vector as a expression vector, make MDR1 gene transfer to the cord blood nucleated cells to explore the gene theropy that can prevent hematopoietic cells from chemotherapeutic drugs attack, let tumor patient can receive large dose chemotherapeutic drugs, enhance the efficiency of chemotherapy and protect hematopoietic cells. Chemotheropy is one of the main treatment method for cancer. but, during chemotherapy, chemotherapeutic drugs usually damaged peripheral blood cells and marrow hematopoietic cells, make the chemotherapy project can't practice according. the schedule. Therefor affact the result of tumor chemotherapy. at present, people offen use hematopoietc growth factor to shorten the time of WBC decrease after chemotherapy, and use composition blood transfusion to overcome the marror suppression after chemotherapy. but they can't prevent marrow suppression from chemotherapy, and reapted chemotherapy can cause stored normal marrow hematopoietic cells decrease, result in marrow hematopoietic fuction failure. One of the method to overcome these question is transfer MDR1 gene into the hematopoietic cells and make it over espression in hematopoietic cells.then make chemotherapy can progress repeatedly in large dose chemotherapeutic drugs. In this study, we utilize resistace of MDR1, advantage of cord blood and character of adenovirus, use adenovirus as a expression vector, make MDR1 gene transfered cord blood nucleated cells (CBNC), enhance it resistance to chemotherapeutic drugs. we hope it can overcome the damage of WBC and marrow hematopoietic during chemotherapy and improve the chemotherapy efficiency. Umbilical Cord Blood is a good substance for human, because it have a extensive source, collects convenience, contains the deal abundant hematopoietic cells, immunogen is weak, no damage to donor. so resent years domistic and international reasercher have a high value to it. it have been used to replace marrow to treatment various blood disease and hereditary disease successfully, also used as a assistance treatment method after chemotherapy, acquired good result. because of its many advantage, it be used to as a target for gene therapy recently. In this reseach, we use advantage of umbilical cord blood, select cord blood nucleated cells (CBNC) as target cells, and utilize adenovirus vector as expression vector, transfer the MDR1 gene into CBNC, make CBNC with MDR1 gene resistance chemotherapeutic drugs. we have finished work include: 1) obtained full lenth of MDR1 gene: isolate the MDR1 gene from PHA MDR plasmid, identified and recovered MDR1 gene by electrophoresis. 2) use adenovirus expression vector transfer the MDR1 gene to the cord blood nucleated cells. 3) extract the DNA of CBNC of transferred MDR1 gene and expand MDR1 gene by PCR in MDR1 full lenth primer, confirm the efficiency of transfer. 4) demonstrate multidrug resistance of CBNC with MDR1 gene by anti-chemotherapeutic drug trial. we can confirm that MDR1 gene have transfered CBNC from the result of PCR expand, therefore, resistance trial for CBNC with MDR1 gene also confirm that CBNC with MDR1 gene have a obvious resistance to chmotherapeutic drug VCR (P<0.01, compared with control group), and two time transferred CBNC have a obviously resistance to VCR than one time transferred CBNC. The result of research demonstrate that CBNC is a good target for gene trans...
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