摘要
目的 探索钙调素与细胞增殖及肿瘤发生发展的关系。方法 从全血分离的淋巴细胞以 5× 10 5/ml接种于含10 %小牛血清的RPMI16 40中 ,加入PHA 2 0mg/L使其活化 ,以不加PHA者为对照 ;K5 6 2细胞以 2× 10 5/ml接种 ,加入CaM拮抗剂TFP 2 0 μmol/L ,以不加TFP组为对照。上述细胞于 37℃、5 %CO2 培养箱中培养 ,离心收集细胞 ,提取总RNA ,以管家基因 β 肌动蛋白 (β actin)为内参照 ,用RT PCR方法检测细胞内CaMmRNA的表达水平。结果 PHA活化的淋巴细胞和K5 6 2细胞CaMmRNA的表达水平分别为1 184± 0 0 2 3和 2 740± 0 132 ,均显著高于静息淋巴细胞(0 5 75± 0 15 3) (P <0 0 1) ,与TFP共育不影响K5 6 2细胞CaMmRNA的表达水平。结论 胞内CaM与细胞增殖有关 。
Objective To evaluate association of CaM with cell proliferation and tumor genesis. Methods Quiescent l ymphocytes were obtained from healthy donors and cultured at 5×105/ml in RPMI 1640 medium supplemented with 10% new born calf serum, and with the presence or absence of PHA(20 mg/L). K562 cells were seeded at 2×105/ml, and CaM antagon ist TFP was added to 20 μmol/L,while K562 cells not treated with TFP were used a s control. After being cultured under 37 ℃ in a humidified atmosphere containin g 5% CO 2.cells were collected by centrifugation and total cellular RNA was ext racted. Levels of intracellular CaM mRNA were evaluated by RT-PCR using houseke eping gene β-actin as a reporter gene.Results Levels of CaM m RNA in PHA-activated lymphocytes and K562 cells were 1.184±0.023 and 2.740±0 132 respectively, significantly higher than those of quiescent lymphocytes (0. 575±0 153)( P <0 01). Moreover,it was found that levels of CaM mRNA were n ot affected when K562 cells were incubated with TFP.Conclusion CaM plays a role in cell proliferation and may be involved in the genesis and de velopment of tumor
出处
《肿瘤》
CAS
CSCD
北大核心
2001年第4期263-265,共3页
Tumor
基金
江苏省自然科学基金资助 (编号 :BK95 14 13 0 7)
关键词
钙调蛋白
MRNA
细胞分裂
淋巴细胞
肿瘤细胞
Calmodulin
RNA
Neoplasms
Cell divisi on
Lymphocytes
Tumor cells,cultured
