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用快速印迹(quick blot)和延续转录法(run-offtranscription assay),证明在1-o-十四酰基咐(口拜)醇-13-乙酸酯(12-o-tetradccanoyl phorbol-13-acctate,TPA)刺激下,静止期NIH 3T3细胞中c-fos,c-myc,c-myb表达均有增加,但在蛋白激酶C(protcin kinase c,PK—C)抑制剂十六酰基肉毒碱(pamitoyl-L-carnitinc chloride,PMC)存在下,TPA诱发的c-fos,c-myc,c-myb表达均受到抑制。说明促癌剂TPA的基因诱导作用是通过PK-C介导的。c-fos等初级反应基因表达的检测和PK-C在这些基因表达中的作用分析有希望作为TPA型促癌物的检测方法。
Using a quick blot and a run-off transcription assay, it was demonstrated in 1-o-tetradccanoyl phorbol-13-acctate. The expression of c-fos, c-myc, and c-myb in quiescent NIH 3T3 cells was increased under stimulation with TPA, but in the inhibitor of protein kinase C (PK-C) hexadecanoyl carnitine In the presence of pamitoyl-L-carnitinc chloride (PMC), the expression of c-fos, c-myc, c-myb induced by TPA was inhibited. This indicates that the gene-inducing effect of the promoter TPA is mediated by PK-C. The detection of the expression of primary response genes such as c-fos and the analysis of the role of PK-C in the expression of these genes are promising as a method for the detection of TPA-type carcinogens.