EFFECT OF MICROCYSTIN LR ON PP2A IN HL7702 CELL

L. Jing
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Abstract

Microcystins(MCs) are hepatotoxins which inhibit the activity of protein phosphatase 2A(PP2A) notably.The cells were incubated with different concentrations of MC-LR to determine the effect of MCs on PP2A in HL7702 respectively for 24h.The mRNA and protein levels of PP2A subunits were detected.Furthermore,the activity of PP2A was assayed after 3h,6h,12h and 24h exposure.The protein levels were not changed significantly except B55α which decreased concentration-dependently and significantly in 1000 nmol/L.For mRNA levels,15 subunits increased in 100 nmol/L group,whereas 7 subunits in 500 nmol/L group and 6 subunits in 1000 nmol/L group decreased.The activity of PP2A was inhibited significantly and concentration-dependently after 6h exposure while there were no changes in 3,12 and 24h exposure.In conclusion,MC-LR affects the transcription of PP2A subunits after 24h exposure,without concomitant changes in the interrelated proteins.Exposure time might be a determinant when considering the effect of MC-LR on PP2A activity.
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微囊藻毒素lr对hl7702细胞pp2a的影响
微囊藻毒素(microcystiins, MCs)是一种显著抑制蛋白磷酸酶2A(PP2A)活性的肝毒素。用不同浓度的MC-LR分别孵育细胞24h,测定mc对HL7702中PP2A的影响。检测PP2A亚基mRNA和蛋白水平。此外,在暴露3h、6h、12h和24h后检测PP2A的活性。除B55α在1000 nmol/L浓度下呈浓度依赖性显著降低外,其余蛋白水平无显著变化。在mRNA水平上,100 nmol/L组有15个亚基升高,500 nmol/L组有7个亚基降低,1000 nmol/L组有6个亚基降低。暴露6h后,PP2A活性明显受到抑制,且呈浓度依赖性,而暴露3、12、24h时,PP2A活性无明显变化。综上所述,MC-LR在暴露24h后会影响PP2A亚基的转录,而相关蛋白没有随之改变。在考虑MC-LR对PP2A活性的影响时,暴露时间可能是一个决定因素。
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