v-src,Ca^(2+),PKC和cAMP对3T3-tsLA90细胞间隙连接通讯的调节  

THE REGULATION OF v-src GENE,CALCIUM,CALMODULIN,PROTEIN KINASE C AND cAMP ON THE JUNCTIONAL INTERCELLUAR COMMUNICATION IN 3T3-tsLAg0 CELLS

作　　者：张云[1] 王端顺[1] 张鸿卿[1] 王瑞虹[1] 薛绍白[1] 

机构地区：[1]北京师范大学生物系,北京新外大街100875

出　　处：《北京师范大学学报（自然科学版）》1992年第2期233-239,共7页Journal of Beijing Normal University(Natural Science)

基　　金：国家自然科学基金

摘　　要：以3T3-tsLA 90细胞为正常和转化细胞模型,采用划痕标记染料示踪技术,初步研究了 v-src 基因及 Ca^(2+)-钙调素,蛋白激酶 C,cAMP 等细胞内信号物质对细胞间隙连接通讯的调节作用.40℃时正常表型的细胞间隙连接通讯正常,33℃时转化表型的细胞间隙连接被阻断.用 v—src 基因产物抑制剂、钙通道抑制剂、钙离子载体、钙调素和蛋白激酶 C 抑制剂及 cAMP研究了对细胞间隙连接的作用.当3T3-tsLA 90细胞由40℃转移到33℃时,v-src 基因活化,其产物 pp 60^(v-s c)通过 Ca^(2+)-CaM 和 DG-PKC 通路及 cAMP 系统相互作用对细胞间隙连接通讯功能起下降调节作用,从而导致细胞呈转化表型状态.Using 3T3-tsLA 90 cells as normal and its transformed cell medols,the regulatory effects of v-src gene,calcium,calmodulin,PKC and cAMP on intercellular junctional communication are investigated with scrape-loading and dye transfer technique. It is observed that intercelluar junction of 3T3-tsLA 90 cells cultured at the nonpermissive temperature 40 ℃ are permeable by the dye Lucifer yellow CH,that junctional permeability fall after the temperature is shifted to the permissive 33 ℃ for 1 h and that such fall in junctional permeability could be prevented by pretreating the cells with the inhibitor of pp60^(v-src)quercetin for 2h before the temperature shifiting.These facts indicate that the junctional alteration is a primary event in the transfor- mation process.it might be one of the earliest signal of transformal phe- notype and the state of gap junction of these cells is controled by v-src gene.It is also found that the downregulation effect of pp^(60v-src)at 33 ℃ on junctional communication could be mimicked by supplying the culture at nonpermissive temperature 40 ℃ with either calcium ionophore A23187-Br or PKC activator TPA,conversely,inhibition of calcium-CaM system by application of verapamil plus TWB-8(the inhibitors of exogenous and endog- enous Ca_2^+,respctively),TFP/W7(inhibitors of CaM)or inhibition of PKC by H7 all blocked the pp 60^(v-src)downregulation effect on junctional permeability at 33 ℃.The downregulation of junctional communication of 3T3-tsLA 9O cells is inhibited by both sodium butyrate and dibutyryl-cAMP that shows the increase in cAMP level inhibit the fall of junctional perme- ability in 3T3-tsLA 90 cells.These results reveal that Ca_2^+-CaM and DG- PKC system are critically involved in the route of pp 60^(v-src)taking to exert its downregulating effect on junctional permability.

关 键 词：间接连接通讯 细胞间隙 PKC 钙调素 

分 类 号：Q26[生物学—细胞生物学]