Sulfatase 2 protects hepatocellular carcinoma cells against apoptosis induced by the PI3K inhibitor LY294002 and ERK and JNK kinase inhibitors.
Sulfatase 2 protects hepatocellular carcinoma cells against apoptosis induced by the PI3K inhibitor LY294002 and ERK and JNK kinase inhibitors.
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DOI：
10.1111/j.1478-3231.2010.02336.x
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发表时间：
2010-11
期刊：
影响因子：
--
通讯作者：
Roberts LR
中科院分区：
文献类型：
--
作者：
Lai JP;Sandhu DS;Yu C;Moser CD;Hu C;Shire AM;Aderca I;Murphy LM;Adjei AA;Sanderson S;Roberts LR
硫酸酯酶2（SULF 2）是一种细胞外硫酸乙酰肝素6-O-内硫酸酯酶，在肝细胞癌（HCC）中具有致癌作用，部分通过磷脂酰肌醇蛋白聚糖3介导，其促进肝素结合生长因子信号传导和HCC细胞生长。SULF 2还增加抗凋亡Akt激酶底物GSK 3 β的磷酸化，并且SULF 2表达与人HCC中凋亡指数降低相关。我们采用免疫组化、Western免疫印迹、基因转染、实时定量聚合酶链反应、MTT和凋亡检测以及免疫细胞化学等方法研究了SULF 2对药物诱导的肝癌细胞凋亡的功能和机制。通过免疫组化和免疫印迹证实SULF 2在人HCC中的表达增加。用MEK、JNK和PI 3激酶抑制剂处理降低了SULF 2阴性Hep 3B HCC细胞的活力，并诱导凋亡的半胱天冬酶3和7活性，其中PI 3 K抑制剂LY 294002诱导的凋亡最强。SULF 2在Hep 3B细胞中的强制表达显著降低了凋亡半胱氨酸蛋白酶3和7的活性，并诱导对LY 294002诱导的凋亡的抵抗。如预期的，LY 294002抑制PI 3 K对Akt激酶的激活。相反，使用靶向SULF 2 mRNA的shRNA构建体敲低SULF 2诱导了显著的细胞生长停滞，并使内源性表达SULF 2的HCC细胞系Huh 7和SNU 182对药物诱导的凋亡敏感。SULF 2基因敲低对肝癌细胞的影响是通过Akt磷酸化减少、细胞周期蛋白D1和抗凋亡分子Bcl-2下调以及促凋亡分子BAD上调介导的。SULF 2在HCC中的促存活、抗凋亡作用是通过激活PI 3 K/Akt通路介导的。
Sulfatase 2 (SULF2), an extracellular heparan sulphate 6-O-endosulphatase, has an oncogenic effect in hepatocellular carcinoma (HCC) that is partially mediated through glypican 3, which promotes heparin-binding growth factor signalling and HCC cell growth. SULF2 also increases phosphorylation of the anti-apoptotic Akt kinase substrate GSK3β and SULF2 expression is associated with a decreased apoptotic index in human HCCs. We investigated the functional and mechanistic effects of SULF2 on drug-induced apoptosis of HCC cells using immunohistochemistry, Western immunoblotting, gene transfection, real-time quantitative polymerase chain reaction, MTT and apoptosis assays and immunocytochemistry. The increased expression of SULF2 in human HCCs was confirmed by immunohistochemistry and immunoblotting. Treatment with inhibitors of MEK, JNK and PI3 kinases decreased the viability of SULF2-negative Hep3B HCC cells and induced apoptotic caspase 3 and 7 activity, which was most strongly induced by the PI3K inhibitor LY294002. Forced expression of SULF2 in Hep3B cells significantly decreased activity of the apoptotic caspases 3 and 7 and induced resistance to LY294002-induced apoptosis. As expected, LY294002 inhibited activation of Akt kinase by PI3K. Conversely, knockdown of SULF2 using an shRNA construct targeting the SULF2 mRNA induced profound cell growth arrest and sensitized the endogenously SULF2-expressing HCC cell lines Huh7 and SNU182 to drug-induced apoptosis. The effects of knockdown of SULF2 on HCC cells were mediated by decreased Akt phosphorylation, downregulation of cyclin D1 and the anti-apoptotic molecule Bcl-2, and upregulation of the pro-apoptotic molecule BAD. The prosurvival, anti-apoptotic effect of SULF2 in HCC is mediated through activation of the PI3K/Akt pathway.
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影响因子：
25.7
作者：
Lai JP;Sandhu DS;Moser CD;Cazanave SC;Oseini AM;Shire AM;Shridhar V;Sanderson SO;Roberts LR
通讯作者：
Roberts LR
影响因子：
11.2
作者：
Chau, B. Nelson;Diaz, Robert L.;Cleary, Michele A.
通讯作者：
Cleary, Michele A.
影响因子：
4.8
作者：
Dai, YM;Yang, Y;Sanderson, RD
通讯作者：
Sanderson, RD
影响因子：
11.2
作者：
Chen, Kuen-Feng;Yeh, Pei-Yen;Cheng, Ann-Lii
通讯作者：
Cheng, Ann-Lii
影响因子：
13.5
作者：
Lai, Jin-Ping;Sandhu, Dalbir S.;Roberts, Lewis R.
通讯作者：
Roberts, Lewis R.