论文部分内容阅读
目的:研究温石棉对人胸膜间皮细胞MeT-5A细胞毒性与诱导细胞恶性转化的能力。方法:于2016年6月，采用乳酸脱氢酶（LDH）法检测温石棉对MeT-5A细胞的细胞毒性；以5 μg/cmn 2温石棉间断染毒MeT-5A细胞，24 h/次，每周一次，共28次，待细胞呈现锚着不依赖性生长后，用软琼脂集落形成试验判断细胞恶性转化，并计算软琼脂集落形成率，紫外比色法测定糖代谢过程中的关键限速酶活性，包括丙酮酸激酶（PK）、磷酸果糖激酶（PFK）和己糖激酶（HK）。n 结果:温石棉对MeT-5A细胞具有细胞毒性，细胞相对存活率呈浓度依赖性下降。与未染毒时相比，10、20、40、80 μg/cmn 2温石棉染毒后MeT-5A细胞相对存活率显著下降（n P<0.05）。染毒28次后，温石棉转化组细胞生长速度加快、排列紊乱，失去接触抑制，出现叠层生长，并可在软琼脂上生长，细胞集落形成率为18.33‰±2.49‰，与对照组（0）比较，差异有统计学意义（n P<0.01）；温石棉转化组细胞PK[（19.51±1.52）U/L]、PFK[（0.12±0.02）U/10n 4个细胞]和HK[（0.26±0.01）U/10n 4个细胞]酶活性水平高于对照组细胞[（25.00±1.04）U/L、（0.15±0.01）U/10n 4个细胞和（0.33±0.01）U/10n 4个细胞]（n P<0.01）。n 结论:温石棉可诱导MeT-5A细胞发生恶性转化，并升高细胞糖酵解相关激酶PK、PFK、HK酶活性水平。“,”Objective:To study the cytotoxicity and malignant transformation ability of chrysotile on MeT-5A cells.Methods:In June 2016, lactate dehydrogenase (LDH) method was used to detect the cytotoxicity of chrysotile to MeT-5A cells. MeT-5A cells were treated with 5 μg/cm n 2 chrysotile intermittently for 24 h a time, once a week and a total of 28 times. After the cells showed anchorage independent growth, the cell features of malignant transformation were identified by colony forming frequency in soft agar, and the soft agar colony formation rates were calculated. The activities of key speed limiting enzymes of glycolysis metabolism including hexokinase (HK) , phosphofructokinase (PFK) and pyruvate kinase (PK) were determined by UV colorimetry.n Results:Chrysotile was cytotoxic to MeT-5A cells in a concentration-dependent decline. Compared with the control group, the relative survival rates of MeT-5A cells were significantly decreased after exposed to chrysotile at 10, 20, 40 and 80 μg/cm n 2 (n P<0.05) . After 28 times of exposure, the growth rate of the cells in chrysotile transformed MeT-5A cells was accelerated, the arrangement was disordered, the contact inhibition was lost, and the double layer growth appeared, which could grow on soft agar. The colony forming rate of the chrysotile transformed MeT-5A cells was 18.33‰±2.49‰. Compared with the control group (0) , the difference was statistically significant (n P<0.01) . The activities of glycolysis related kinase including PK [ (19.51±1.52) U/L], PFK[ (0.12±0.02) U/10n 4 cell] and HK[ (0.26±0.01) U/10n 4 cell] were increased in the chrysotile transformed MeT-5A cells compared with control group [ (25.00±1.04) U/L、(0.15±0.01) U/10n 4 cell and (0.33±0.01) U/10n 4 cell] (n P<0.01) .n Conclusion:Chrysotile can induce malignant transformation of MeT-5A cells and increase the activities of glycolysis related kinases including PK, PFK and HK.