应用荧光DDRT-PCR初探低剂量的ECPs诱导细胞适应性反应的机理

RESEARCH ON MECHANISM OF CELL ADAPTIVE RESPONSE INDUCED BY LOW CON-CENTRATION OF ECPS USING FLUORESCENCE DDRT-PCR

[目的]用荧光DDRT-PCR技术寻找低剂量环境化学污染物(ECPs)过氧化氢(H_2O_2)、甲醛(FA),三氯乙烯(TCE)诱导细胞适应性反应差异表达的基因,为进一步研究低剂量的环境化学污染物(ECPs)诱导机体的生物学效应及其机理提供科学依据。[方法]依据H_2O_2、FA、TCE对细胞毒性的剂量反应关系,建立适应性反应的模型。然后用荧光DDRT-PCR寻找对照组、低剂量组、高剂量组,适应性反应组差异表达的基因,并鉴定和验证部分差异表达的基因。[结果]按照对照组、低剂量组、高剂量组、适应性反应组处理细胞后,DDRT-PCR结果显示各处理组与对照组比较:H_2O_2有60个差异条带、FA有61个,TCE有52个。选择差异比较明显的条带进行2次PCR,然后通过进一步克隆、测序、同源性比较发现一些已知基因如BCL-2、EIF3S5、NDUFS4、RPS10等。[结论]低剂量环境化学污染物诱导机体的适应性反应可能有维持细胞稳态、抗氧化系统、DNA损伤修复级激活细胞信号分子等过程的参与。

[Objective] To observe differential display genes of cell adaptive response induced by low concentration of environmental chemical pollution substance （ECPs） , such as hydrogen peroxide （H2Oz） , formaldehyde （FA） and trichloroethylene （TCE） and to explore the mechanism of adaptive response. [Methods] Models of adaptive response of cells induced by hydrogen peroxide （H2O2）. formaldehyde （FA）. triehloroethylene （TCE） were established according to the doseeffect manners of their toxicology, then differential display genes of different treatment groups were determined using fluorescence differential display polymerase chain reaction （DDRT-PCR） . [Results] The results showed that 60 differential display straps of H2O2, 61 FA, 52 TCE were found among low dose group, high dose group, and adaptive response group compared with control group. Obvious differential display straps were chosen to identified as BCL-2. EIF3S5. NDUFS4. RPS10 and so on. [Conclusion] The mechanism of adaptive response induced by low concentrtration of ECPs maybe include protein of maintaining homeostasis, antioxidation system, DNA damage-repair system, signaling molecules and so on.