阴离子交换蛋白1与P16蛋白在HeLa细胞中共表达

Co-expression of anion exchanger 1 and P16 in HeLa cells

目的在人宫颈癌细胞株(HeLa)中共表达具有阴离子转运活性的阴离子交换蛋白(anion exchanger 1,AE1)与P16蛋白,观察两者之间的相互作用。方法用脂质体将目的基因转入HeLa,Western blot及免疫荧光方法验证蛋白表达及定位,描绘细胞生长曲线,运用6-甲氧基-N-(3-磺丙基)喹啉铵(SPQ)测定细胞阴离子转运活性。结果 P16和AE1均成功在HeLa细胞中过表达。共同转染pEGFP-C1-p16和pRBG4-AE1后HeLa细胞的阴离子交换功能明显增强,且细胞增殖能力明显下降。结论 P16与AE1之间存在相互作用和/或协同作用,为进一步研究AE1在细胞增殖和肿瘤发生、发展过程中作用奠定了基础。

Objective To investigate the interaction between P16 and anion exchanger 1（AE1）,a protein with anion transporting activity,in a cervical cancer cell line named HeLa constructed with co-expression of both proteins.Methods AE1 and P16 expression vector were co-transfected into HeLa cells with LipofectamineTM2000.Immunofluorescence and Western blot were employed to verify the protein expression and localization.The effects on cell growth were evaluated by cell growth curve.SPQ（6-methoxy-N-（3-sulfopropyl）-quinolinium） was used to detect the anion transporting activity of cells.Results Successful expression of P16 and AE1 were verified in HeLa.Cells with co-expression of pEGFP-C1-p16 and pRBG4-AE1 showed significant enhancement in anion transporting activity and reduction in proliferation ability.Conclusion These results demonstrate the interaction and cooperative role of tumor suppressor AE1 with P16.Also,it provides a novel regulation pathway for both AE1 and P16.