五氯酚对HeLa细胞毒性及DNA损伤的研究

以人宫颈癌HeLa细胞为研究对象,运用MTT法检测PCP处理后HeLa细胞的增长抑制率;通过测定细胞培养液中乳酸脱氢酶(LDH)的漏出率和总超氧化物歧化酶(SOD)的活性,评价PCP的细胞毒性作用;彗星实验检测HeLa细胞经不同浓度PCP处理后的DNA损伤.结果表明,PCP对HeLa细胞的半数抑制浓度(IC50)为66.59μmol.L-1;HeLa细胞在12.5、25、50、100和200μmol.L-1 PCP染毒条件下,LDH的漏出率随着染毒时间的增加逐渐增大,具有明显的时间-效应关系,在12.25、17.5和25μmol.L-1 PCP染毒下,细胞培养液中SOD的活性随着染毒时间的增加逐渐下降,存在显著的时间-效应关系,低浓度PCP(25μmol.L-1)显著增加细胞培养液中LDH的漏出率以及PCP(12.25μmol.L-1)显著降低总SOD的活性;PCP(实验浓度为6.25、12.5、25和50μmol.L-1)不会导致HeLa细胞DNA损伤.因此SOD和LDH可作为评价低浓度PCP毒性效应的敏感性生物标志物.

Effects of Pentachlorophenol on DNA Damage and Cytotoxicity of HeLa Cells

Using human cervical carcinoma HeLa cells, the cell viability was determined by MTT assay after pentachlorophenol (PCP) treatment, the cytotoxicity of PCP was evaluated by detecting lactate dehydrogenase (LDH) leakage rate and total superoxide dismutase (SOD) activity in cell culture medium; DNA damage was detected by comet assay. The results indicated that HeLa cells proliferation was inhibited by PCP and the median inhibitory concentration (IC50) was 66.59 micromol x L(-1); PCP did not induce DNA damage in the concentration range from 6.25 micromol x L(-1) to 50 micromol L(-1); LDH leakage rate increased gradually with the increasing of exposure time when HeLa cells were treated by PCP in the concentration range from 12.5 micromol x L(-1) to 200 micromol x L(-1); SOD activity decreased gradually as the increasing of exposure time when HeLa cells were treated by PCP at lower concentration of 12.25 micromol x L(-1), 17.5 micromol x L(-1), 25 micromol x L(-1) respectively, LDH leakage rate increased significantly at 25 micromol x L(-1) and activity of SOD decreased markedly at 12.25 micromol x L(-1) in HeLa cells following PCP-treatment respectively. Results suggested that SOD and LDH might be regarded as candidate sensitive biomarkers for evaluating toxicity of PCP at low concentration on human and wildlife.