【作者】 陈文芳；

【导师】 齐浩；

【作者基本信息】 陕西师范大学 ， 动物学， 2005， 硕士

【摘要】 目的:研究磁场对细胞增殖的影响及其可能的机理。将磁场结合不同药物作用于肿瘤细胞,研究磁场与药物的协同抗癌作用。 方法:1.采用MTT法检测磁场对肿瘤细胞(K562和SMMC—7721细胞)和原代乳鼠肝细胞增殖的影响。2.通过HE染色、扫描电镜、原子力显微镜、透射电镜的观察和细胞周期分布及胞内游离钙离子浓度的检测,研究磁场抑制K562细胞增殖的可能原因。3.利用MTT法检测磁场结合环磷酰胺对K562细胞增殖的影响,并通过透射电镜观察和细胞周期分布检测探索磁场协同环磷酰胺抗肿瘤作用的可能原因。4.用MTT法检测磁场结合中药(土贝母皂甙、白屈菜红碱和血根碱)对K562细胞的杀伤作用。 结果:1.K562细胞以5×10~4cells/mL的细胞密度接种,经9mT磁场处理0～72h,MTT检测的结果表明曝磁12h时细胞增殖即受到抑制(P<0.05);以1×10~5cells/mL的细胞密度接种,经磁场处理0～24h,结果表明曝磁24h时细胞增殖受到抑制(P<0.05)。SMMC—7721细胞以5×10~4cells/mL的密度接种,经磁场处理0～72h,结果表明9h时细胞增殖即受到抑制(P<0.05)。原代乳鼠肝细胞以2×10~5cells/mL的细胞密度接种,经磁场处理0～36h,结果显示磁场在0～24h时对细胞增殖没有影响(P>0.05),暴露至36h时,曝磁组细胞的活性显著高于对照组(P<0.05)。 2.K562细胞经磁场处理后,细胞形态改变,细胞表面出现突起,细胞膜不完整,胞质内空泡增多,线粒体膨胀,染色质凝集,核膜增厚,部分细胞解体。磁场处理后K562细胞的周期分布改变。在0～24h内,随曝磁时间增加,G1、S期细胞减少,G2/M期细胞比例升高。比率荧光法检测胞内钙离子浓度的结果显示曝磁组细胞的胞内钙离子浓度从曝磁6h后即低于对照组细胞。 3.K562细胞经磁场与环磷酰胺联合处理12h时,MTT检测即发现细胞活性显著降低,与单纯环磷酰胺处理的细胞相比有显著性差异(P<0.01),联合处理24h后,细胞活性下降更为明显。细胞周期分析结果表明环磷酰胺结合磁场处理细胞12h后,S期细胞降低,G2/M期细胞上升;处理24h后,大部分细胞处于S期。透射电镜观察发现磁场结合环磷酰胺处理K562细胞24h后,细胞胞质内出现数量不等的空泡,有许多电子密度较高的颗粒状或泡状结构,并观察到有核膜增厚的现象。 4.K562细胞经土贝母造甙、白屈菜红碱、血根碱分别结合磁场处理24h后,MTT检测结果表明细胞活性均显著下降,与各自的单纯药物处理组相比,均有显著性差异(P<0.01)。更多还原

【Abstract】 Aim: We studied the growth inhibitory effects of static magnetic fields (SMF) on cancer cells, and the synergistic anticancer effects between static magnetic fields and drugs.Methods: 1. the survival of human tumor cells (K562 cells and SMMC-7721 cells) and rodent hepatocytes treated by SMF were measured by MTT test. 2.The morphologic changes of K562 cells exposed under SMF were observed by microscopy, scanning electron microscopy (SEM), atomic force microscopy (AFM) and transmission electron microscopy (TEM); The cell cycle distribution and cytoplasmic calcium concentration ([Ca2+]i)of K562 cells after exposure were tested by flow cytometer (FCM) and ratio fluorescence measurement, respectively. 3. The influence of SMF combined with cyclophosphamide (CP) on K562 cells were detected by MTT test, TEM and FCM. 4. The kill effects of SMF cooperated with Chinese herbs on K562 cells were tested by MTT test.Results: 1.When cell density was 5×10~4cells/mL, the growth of K562 cells were inhibited after exposing under SMF for 12h (P<0.05) .While the cell density was 1 × 10~5 cells/mL ,the growth of K562 cells were supressed after exposing under SMF for 24h (P<0.05) . The proliferation of SMMC-7721 cells were declined after exposing under SMF for 9h (P<0.05) . There were no influence on growth of rodent hepatocytes which was treated under SMF for 24h (P>0.05) .2. After exposure, there were some changes in the morphology of K562 cells, such as some protuberances appeared on the cell membrane, mitochondria swelled, and nuclear membranes incrassated. When K562 cells were treated with SMF for 24h, the cell cycle distributions would changed, and ratio of cells at G1 and S phase was down, while those at G2/M phase was up. The cytoplasmic calcium concentrations of exposed K562 cells were lower than those of controls after 6h.3. When K562 cells were dealt with SMF and CP for 12h, the cell activities were significantly reduced (P<0.01), and the cell cycle distributions as well as ultrastructure were also changed, compared with controls.4. When K562 cells were treated with SMF cooperated with tubeimuside, chelerythrine or sanguinarine respectively for 24h, cell activities were significantly declined.更多还原