论文部分内容阅读
AIM We have previously reported that inducibleover-expression of Bak may prolong cell cycle inG_1 phase and lead to apoptosis in HCC-9204 cells.This study is to investigate whether p27~(KIP1)playsan important role in this process.METHODS In order to elucidate the exactfunction of p27~(KIP1)in this process,a zinc induciblep27~(KIP1)stable transfectant and transient p27~(KIP1)-GFP fusion transfectant were constructed.Theeffects of inducible,p27~(KIP1)on cell growth,cellcycle arrest and apoptosis were examined in themock,control pMD vector,and pMD-KIP1transfected HCC-9204 cells.RESULTS This p27~(KIP1)-GFP transfectant maytransiently express the fusion gene.The cellgrowth was reduced by 35% at 48 h of p27~(KIP1)induction with zinc treatment as determined bytrypan blue exclusion assay.These differencesremained the same after 72 h of p27~(KIP1)expression,p27~(KIP1)caused cell cycle arrest after24 h of induction,with 40% increase in G_1population.Prolonged p27~(KIP1)expression in thiscell line induced apoptotic cell death reflected byTUNEL assay.Fourty-eight h and 72 h of p27~(KIP1)expression showed a characteristic DNA ladder onagarose gel electrophoresis. CONCLUSION Bak may induce cell cycle arrest inG_1 phase through upregulating expression ofp27~(KIP1)and subsequently lead to apoptosis inHCC-9204 cells.The p27~(KIP1)-GFP fusion proteincan be transiently expressed in HCC-9204 cells.The inducible p27~(KIP1)-expressing cell line providesa model to assess p27~(KIP1)function.
AIM We have previously reported that inducibleover-expression of Bak may prolong cell cycle inG_1 phase and lead to apoptosis in HCC-9204 cells.This study is to investigate whether p27~(KIP1)playsan important role in this process.METHODS In order to elucidate The exact function of p27~(KIP1) in this process,a zinc induciblep27~(KIP1)stable transfectant and transient p27~(KIP1)-GFP fusion transfectant were constructed.The effects of inducible,p27~KIP1) on cell growth,cell arrest And apoptosis were examined in themock,control pMD vector,and pMD-KIP1transfected HCC-9204 cells.RESULTS This p27~(KIP1)-GFP transfectant maytransiently express the fusion gene.The cellgrowth was reducedat 35% at 48 h of p27~( KIP1)induction with zinc treatment as determined bytrypan blue exclusion assay.The difference between the effects of 72h of p27~KIP1 expression, p27~KPI1caused cell cycle arrest after24h of induction, with 40% increase in G_1population.Prolonged P27~(KIP1)expression in this cell lin单词induced apoptotic cell death reflected byTUNEL assay.Fourty-eight h and 72h of p27~(KIP1)expression showed a characteristic DNA ladder onagarose gel electrophoresis. CONCLUSION Bak may induce cell cycle arrest inG_1 phase through upregulating expression ofp27~(KIP1)and Then lead to apoptosis in HCC-9204 cells. The p27~(KIP1)-GFP fusion protein can be transiently expressed in HCC-9204 cells. The inducible p27~(KIP1)-expressing cell line providesa model to assess p27~(KIP1)function.