转基因CHO细胞中VMAT_2抗毒性作用的研究

Study on the mithridatism of VMAT_2 in transgeneic CHO cell

目的 研究转基因中国仓鼠卵 (CHO)细胞中单胺囊泡转运体 (VMAT2 )的抗毒性作用。方法 利用转 PC1 2 细胞基因到 CHO细胞中形成的转基因 CHO细胞 (c DNACHO) ,采用 MTT比色法检测 1-甲基 -4 -苯基吡啶离子 (MPP+ )对 CHO细胞野生株 (wt CHO)和 c DNACHO细胞的毒性作用 ,并观察利血平——VMAT2 的特异性阻滞剂对 MPP+毒性作用的影响。结果 在 MPP+ 0 .5 m mol/ L 以上浓度 c DNACHO细胞对 MPP+ 敏感性比 wt CHO低得多 ;c DNACHO和 wt CHO对鱼藤酮 (rotenon)的敏感性无显著差异 ;加入VMAT2 的特异性阻滞剂利血平后 ,上述保护作用消失 ,c DNACHO对 MPP+ 敏感性与 wt CHO细胞无差异 ,而单独予以 wt CHO细胞利血平则不能改变它对低浓度 MPP+的敏感性。结论 此保护机制是由转基因细胞中 VMAT2 引起的 ,VMAT2 在转基因的非神经细胞系 (CHO细胞系 )中也能将 MPP+转运至囊泡内 ,从而保护细胞 ;同时也提示 PC1 2

Objective To study the mithridatism of VMAT 2 in transgeneic CHO cell.Methods Using technology of transgene from PC 12 to CHO, MTT reduction assay was used to detect the toxic effect on MPP + to wtCHO and cDNACHO,meanwhile the role of reserpine was observed,including the toxic effect to MPP + on specific blocking agent of VMAT 2.Results cDNACHO to the sensitivity of MPP + was much less than that of wtCHO over concentration of 0.5 mmol/L MPP +; cDNACHO had the same sensitive as wtCHO to rotenon;after the reserpine was added,the above role disappeared,but wtCHO reserpine was given alone,it couldn't change its sensitivity to MPP +.Conclusion VMAT 2 has protective effect on cDNACHO by transporting MPP + to vesicles; PC 12 possesses the antitoxic components.