人骨髓间充质干细胞对K562细胞增殖和化疗敏感性的影响.docVIP

人骨髓间充质干细胞对K562细胞增殖和化疗敏感性的影响 正文 人骨髓间充质干细胞对K562细胞增殖和化疗敏感性的影响 文1：人骨髓间充质干细胞对K562细胞增殖和化疗敏感性的影响 Influence of Human Mesenchymal Stem Cells on Cell Proliferation and Chemo-seitivity of K562 Cells Abstract This study was aimed to compare K562 cell proliferation，chemo-seitivity and alteration of MDR1 before and after adhesive culture with MSC，so as to evaluate the relatiohip between chemodrug-resistance of leukemia cells and hemopoietic microenvironment. K562 cell cultivated in suspeion and adhesively cultivated with MSC were collected respectively and cell proliferation curves were drawn; the cell cycle was determined by flow cytometry; the effect of chemotherapy on cellular viability and apoptosis of K562 cell was investigated，the MDR1 gene expression was determined by RT-PCR. The results showed that K562 cells adhesively cultivated with MSC were inhibited and cells in G0/G1 increased (P)，cells in S phase decreased (P) and those in G0/G1 increased (P)，compared with that cultivated in process of daunomycin-inducing apoptosis，K562 cell apoptosis in the adhesive culture with MSC was inhibited (P).MDR1 gene expression in K562 cells was not induced or altered by adhesive is concluded that by co-culture of cell-cell contact with MSC，growth suppression and induction of chemo-resistance of K562 cells take mechanism，however，seems not relevant with MDR1. Key words mesenchymal stem cell; K562; drug resistance; apoptosis; MDR1 耐药是根治白血病的最主要障碍，其发生机制及逆转的研究一直是白血病的研究热点，以往人们主要集中在对单个白血病细胞本身的分子生物学改变的研究，并取得了显著成果，但在体内仍难克服耐药的发生。近年来，随着对骨髓造血微环境（hematopoietic microenviroment，HM）的认识不断加深，发现骨髓基质细胞在白血病细胞恶性克隆的增殖、分化和凋亡中起重要作用［1］。而骨髓基质细胞的前体细胞骨髓间充质干细胞（mesenchymal stem cell，MSC）是骨髓造血微环境中基质细胞的主要来源，对正常造血也具有调控作用，但对白血病细胞的调节作用尚不清楚。本研究以人慢性髓细胞白血病K562细胞株为靶细胞，与正常人MSC共培养，观察其对K562细胞增殖和化疗敏感性的影响，以探讨MSC与白血病耐药的关系。 材料和方法 试剂 柔红霉素、阿霉素（DNR、ADM)，均购自Pharmacia公司；DMEM-LG、RPMI 1640培养液购自Gibco公司；FBS 购自Hyclone公司； RT-PCR试剂盒、蛋白酶K、RNA酶及琼脂糖均购自Promega公司； PCR引物由上海生物工程公司合成。TRIZOL购自Invitrogen公司；Percoll(密度 g/ml)，购自Ameham Biosciences公司。碘化丙锭（P