Type II cGMP-dependent protein kinase inhibits epidermal growth factor-induced phosphatidylinositol-3-kinase/Akt signal transduction in gastric cancer cells.
Type II cGMP-dependent protein kinase inhibits epidermal growth factor-induced phosphatidylinositol-3-kinase/Akt signal transduction in gastric cancer cells.
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DOI：
10.3892/ol.2013.1630
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发表时间：
2013-12
期刊：
影响因子：
2.9
通讯作者：
Qian H
中科院分区：
文献类型：
--
作者：
Wu M;Chen Y;Jiang L;Li Y;Lan T;Wang Y;Qian H
我们的前期研究表明，II型cGMP依赖性蛋白激酶（PKG II）通过抑制表皮生长因子（EGF）受体（EGFR）的磷酸化/活化，抑制EGF诱导的MAPK/ERK和MAPK/JNK介导的信号转导。由于EGFR除了MAPK介导的信号转导途径外还介导其他几种信号转导途径，因此本研究旨在研究PKG II是否能够抑制EGF/EGFR诱导的磷脂酰肌醇-3-激酶（PI 3 K）/Akt介导的信号转导。AGS人胃癌细胞系用编码PKG II的cDNA的腺病毒构建体（Ad-PKG II）感染以增加PKG II的表达，并用8-pCPT-cGMP处理以激活酶。Western blotting检测EGFR、PI 3 K、Akt、mTOR和NF-κB等信号转导通路关键组分的磷酸化/活化。Western blotting检测凋亡相关蛋白Bax、Bcl-2、caspase 9和DNA片段因子（DFF）的表达。末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法检测AGS细胞凋亡。结果显示，EGF处理后EGFR、PI 3 K、Akt和mTOR的磷酸化（活化）增加，NF-κB的核定位（活化）增加。EGF处理也减少了AGS细胞的凋亡，增加了抗凋亡蛋白Bcl-2的表达，但对促凋亡蛋白Bax的表达没有影响，并且没有改变caspase 9和DFF的水平。用Ad-PKG II感染AGS细胞并用8-pCPT-cGMP刺激AGS细胞来增加PKG II活性，抑制EGF诱导的EGFR、PI 3 K、Akt、mTOR和NF-κB的活化;引起caspase 9分解（活化）和DFF水平增加;并逆转EGF的抗凋亡作用。结果表明，PKG II也可能抑制EGF诱导的PI 3 K/Akt介导的信号转导途径，并进一步证实PKG II能够阻断EGFR的激活。
Our previous study revealed that Type II cGMP-dependent protein kinase (PKG II) inhibits epidermal growth factor (EGF)-induced MAPK/ERK and MAPK/JNK-mediated signal transduction through the inhibition of the phosphorylation/activation of the EGF receptor (EGFR). As EGFR also mediates several other signal transduction pathways besides MAPK-mediated pathways, the present study was designed to investigate whether PKG II was able to inhibit EGF/EGFR-induced phosphatidylinositol-3-kinase (PI3K)/Akt-mediated signal transduction. The AGS human gastric cancer cell line was infected with adenoviral constructs encoding a cDNA of PKG II (Ad-PKG II) to increase the expression of PKG II, and treated with 8-pCPT-cGMP to activate the enzyme. Western blotting was used to detect the phosphorylation/activation of the key components of the signal transduction pathway, including EGFR, PI3K, Akt, mTOR and NF-κB. The levels of apoptosis-related proteins, including Bax, Bcl-2, caspase 9 and DNA fragment factor (DFF), were also determined by western blotting. Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling staining was used to detect the apoptosis of the AGS cells. The results revealed that EGF treatment increased the phosphorylation (activation) of EGFR, PI3K, Akt and mTOR, and increased the nuclear localization (activation) of NF-κB. EGF treatment also reduced the apoptosis of the AGS cells and increased the expression of the anti-apoptotic protein, Bcl-2, but had no effect on the expression of the pro-apoptotic protein, Bax, and did not alter the levels of caspase 9 and DFF. Increasing the PKG II activity of AGS cells by infecting them with Ad-PKG II and stimulating them with 8-pCPT-cGMP inhibited the EGF-induced activation of EGFR, PI3K, Akt, mTOR and NF-κB; caused an increase in caspase 9 breakdown (activation) and DFF levels; and reversed the anti-apoptotic effect of EGF. The results suggest that PKG II may also inhibit EGF-induced signal transduction of PI3K/Akt-mediated pathways, and further confirm that PKG II is able to block the activation of EGFR.
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影响因子：
7.3
作者：
Rangaswami H;Schwappacher R;Marathe N;Zhuang S;Casteel DE;Haas B;Chen Y;Pfeifer A;Kato H;Shattil S;Boss GR;Pilz RB
通讯作者：
Pilz RB
DOI：
10.1006/bbrc.1996.0477
发表时间：
1996-03-27
影响因子：
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2008-01-01
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DOI：
10.1016/j.bbrc.2010.07.098
发表时间：
2010-08-27
影响因子：
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