摘要
RasG是盘基网柄菌单细胞增殖过程中的关键蛋白,其基因的异常表达常会导致细胞增殖速度的改变。经观察发现,尿囊酸酶基因干扰后的盘基网柄菌(Dictyostelium discoideum)细胞(RNAi-allC)的分裂速度明显快于野生型KAx-3细胞。为探讨RasG及相关蛋白参与盘基网柄菌细胞增殖的调控机制,该文使用实时荧光定量PCR检测受体酪氨酸激酶/Ras途径的disc I、myo I、gef R、rasG、mkk A、mek A及erk A在KAx-3和RNAi-allC细胞中mRNA水平上的表达情况。蛋白免疫印迹检测两细胞中Disc I和RasG的蛋白含量。实时荧光定量PCR结果显示,所有检测的基因在两种类型细胞中的表达均存在差异;相对于KAx-3细胞,RNAi-allC细胞除disc I和rasG显著下调外,其他基因均明显上调。蛋白免疫印迹结果显示,RNAi-allC细胞中Disc I和RasG的蛋白含量明显少于野生型细胞(P<0.05)。这些数据提示,尿囊酸酶基因的干扰引起了RasG和RasG上下游蛋白含量的变化,最终导致细胞周期缩短,细胞增殖加快。说明RasG及相关蛋白可能参与了盘基网柄菌细胞增殖的调控。
RasG is involved in cell proliferation. The abnormal expression of rasG can significantly affect Dictyostelium discoideum cell proliferation. RNAi-allC cells （silence of allantoicase gene） grew more quickly than wild-type cells KAx-3. To study the molecular mechanisim of RasG and its relative proteins during the cell proliferation, the relative genes of RTK/Ras pathway including disc 1, myo I, gef R, rasG, mkk A, mek A and erk A, were compared by real-time fluorescent quantitative PCR （qRT-PCR）. Disc I and RasG proteins were detected by West- ern blot. qRT-PCR results showed that these genes were obviously differentially expressed in the two types of cells. In contrast with KAx-3 cells, the expression of disc I and rasG in RNAi-allC cells was markedly down-regulated, while others increased. Western blot results showed that the expression of Disc I and RasG was significantly lower in RNAi-allC cells than in KAx-3 cells （P〈0.05）. It indicated that the high-speed division rate of RNAi-allC cells was due to the change of RasG and its relative proteins expression. The above results suggest that RasG and its relative proteins may play important roles in the regulation of D.discoideum cell proliferation.
出处
《中国细胞生物学学报》
CAS
CSCD
北大核心
2013年第6期830-835,共6页
Chinese Journal of Cell Biology
基金
国家自然科学基金(批准号:30970316
30670266)资助的课题~~
