通过研究PKB/Akt与Girdin蛋白之间的关系,目的在于揭示Girdin蛋白在PKB/Akt调控小鼠受精卵微丝聚集中的作用。研究中首先结合软件(http://scansite.mit.edu//)预测了PKB/Akt对Girdin蛋白的磷酸化位点,并制定特定位点磷酸化抗体,通过蛋...通过研究PKB/Akt与Girdin蛋白之间的关系,目的在于揭示Girdin蛋白在PKB/Akt调控小鼠受精卵微丝聚集中的作用。研究中首先结合软件(http://scansite.mit.edu//)预测了PKB/Akt对Girdin蛋白的磷酸化位点,并制定特定位点磷酸化抗体,通过蛋白免疫印迹检测经PKB/Akt m RNA或PKB/Akt si RNA处理后的小鼠受精卵中Girdin蛋白磷酸化改变情况。同时检测了磷酸化的Girdin蛋白亚细胞定位及同微丝骨架的定位关系。进一步通过显微注射PKB/Akt m RNA再注射Girdin si RNA检测小鼠受精卵微丝骨架的聚集。结果显示经持续激活型PKB/Akt m RNA、野生型PKB/Akt m RNA处理后的小鼠受精卵中p-Girdin-1 417分布位置更加集中在2-细胞分裂沟处。经野生型和持续激活型PKB/Aktm RNA注射组p-Girdin-1 417表达增强,但是并不影响Girdin蛋白的总的表达。说明si RNA介导的PKB/Akt表达敲低非常明显地降低了Girdin蛋白的磷酸化。激光共聚焦研究显示在Akt m RNA和Girdin si RNA共注射组微丝骨架分布明显散乱。本研究结果充分说明Girdin蛋白在小鼠受精卵中受到PKB/Akt的调节,PKB/Akt通过磷酸化Girdin蛋白改变微丝骨架的聚集。展开更多
Apigenin is a flavonoid widely distributed in fruits and vegetables. It possesses growth inhibitory properties against numerous cancer cell lines. However,the molecular mechanism(s) by which api-genin elicits its effe...Apigenin is a flavonoid widely distributed in fruits and vegetables. It possesses growth inhibitory properties against numerous cancer cell lines. However,the molecular mechanism(s) by which api-genin elicits its effects have not been fully elucidated. Here we studied whether apigenin inhibits growth and induces apoptosis in human gastric carcinoma cells. We showed that the flavonoid inhibited growth of the cells and caused apoptosis,as evidenced by DNA Ladder,cleavage of pro-caspase-3 in a time-dependent manner. Induction of apoptosis was dependent on inhibition of the PKB/Akt activity. We found that while apigenin had no effect on the expression of Akt and Bad,it inhibited specific phosphorylation of the two proteins that are associated with pro-survival mechanisms. We propose that this important flavonoid induces apoptosis in gastric cancer cells by inhibiting Akt activity. Since Akt is often activated in cancers,our findings may have clinical implications.展开更多
文摘通过研究PKB/Akt与Girdin蛋白之间的关系,目的在于揭示Girdin蛋白在PKB/Akt调控小鼠受精卵微丝聚集中的作用。研究中首先结合软件(http://scansite.mit.edu//)预测了PKB/Akt对Girdin蛋白的磷酸化位点,并制定特定位点磷酸化抗体,通过蛋白免疫印迹检测经PKB/Akt m RNA或PKB/Akt si RNA处理后的小鼠受精卵中Girdin蛋白磷酸化改变情况。同时检测了磷酸化的Girdin蛋白亚细胞定位及同微丝骨架的定位关系。进一步通过显微注射PKB/Akt m RNA再注射Girdin si RNA检测小鼠受精卵微丝骨架的聚集。结果显示经持续激活型PKB/Akt m RNA、野生型PKB/Akt m RNA处理后的小鼠受精卵中p-Girdin-1 417分布位置更加集中在2-细胞分裂沟处。经野生型和持续激活型PKB/Aktm RNA注射组p-Girdin-1 417表达增强,但是并不影响Girdin蛋白的总的表达。说明si RNA介导的PKB/Akt表达敲低非常明显地降低了Girdin蛋白的磷酸化。激光共聚焦研究显示在Akt m RNA和Girdin si RNA共注射组微丝骨架分布明显散乱。本研究结果充分说明Girdin蛋白在小鼠受精卵中受到PKB/Akt的调节,PKB/Akt通过磷酸化Girdin蛋白改变微丝骨架的聚集。
基金Supported by Postgraduate Innovation Grants of Harbin Medical University
文摘Apigenin is a flavonoid widely distributed in fruits and vegetables. It possesses growth inhibitory properties against numerous cancer cell lines. However,the molecular mechanism(s) by which api-genin elicits its effects have not been fully elucidated. Here we studied whether apigenin inhibits growth and induces apoptosis in human gastric carcinoma cells. We showed that the flavonoid inhibited growth of the cells and caused apoptosis,as evidenced by DNA Ladder,cleavage of pro-caspase-3 in a time-dependent manner. Induction of apoptosis was dependent on inhibition of the PKB/Akt activity. We found that while apigenin had no effect on the expression of Akt and Bad,it inhibited specific phosphorylation of the two proteins that are associated with pro-survival mechanisms. We propose that this important flavonoid induces apoptosis in gastric cancer cells by inhibiting Akt activity. Since Akt is often activated in cancers,our findings may have clinical implications.
