酸性神经磷脂酶在维生素E琥珀酸酯诱导胃癌细胞凋亡过程中对内质网应激的影响

doi:10.3969/j.issn.1004-616x.2013.05.001

Abstract

Abstract:

OBJECTIVE: To study the effect of acid sphingomyelinase (ASMase) on endoplasmic reticulum stress (ERS) in vitamin E succinate (VES)-induced apoptosis in human gastric cancer cells. METHODS： SGC-7901 human gastric cancer cells were cultured in vitro. The cells were divided into four groups: desipramine (DES) group (cells were treated at 12.5 μmol/L for 2 h)，control group (containing 2% FBS RPMI-1640 culture medium)，VES+ DES group (cells were treated with DES at 12.5 μmol/L for 2 h，then with VES at 20 μg/ml) and VES group (20 μg/ml). ASMase activity detection kit was used to assess the ASMase activity of VES and VES + DES groups at 0，1.5，3 and 6 h. MTT assay was used to measure the growth inhibition at 12，24，48 h. Cell morphology was examined by the inverted microscope after treated for 24 h，and flow cytometry was used to evaluate the apoptosis，then we measured the ERS-related protein expressions of GRP78，GRP94，Perk，p-Perk，Chopand Caspase-4 by Western blot. RESULTS： ASMase activity continuously increased for VES (20 μg/mL) treatment，peaked at 3 h，then gradually decreased. ASMase activity of VES+DES group remained at a low level. There was no significant change in control group and DES group throughout the tests. Cell proliferation in VES group and VES+DES group was obviously inhibited after treatment for 12，24 and 48 h (all P<0.05) and showed a time-effect trend. Cell relative growth rates at different time points in VES+DES group were significantly higher than those in VES group(all P<0.05). The number of dead cells obviously increased in VES group，and the number of dead cells in VES+DES group decreased compared with VES group. Apoptosis rate evaluated by flow cytometry revealed that VES group (39.21%±1.90%) was significantly higher than the control group(3.91%±1.68%) and the DES group(4.07%±1.39%)(P<0.05)，VES+DES group (19.47%±4.46%) was significantly lower than the VES group(P<0.05). ERS-related protein GRP78，GRP94，p-Perk，Chop and c-Caspase-4 expression levels in VES+DES group were lower than those in VES group (all P<0.05). CONCLUSION：VES may induce apoptosis of human gastric cancer cell SGC-7901 cells though ERS by ASMase regulation pathways.

Key words: acid sphingomyelinase, vitamin E succinate, endoplasmic reticulum stress, apoptosis

DONG Rui-hua，ZHAO Wei，WANG Li-ming，ZHANG Xu-guang，WU Kun. Effects of acid sphingomyelinase on endoplasmic reticulum stress in VES- induced apoptosis in human gastric cancer cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, doi: 10.3969/j.issn.1004-616x.2013.05.001.

References

[1] 贾莉，王彦军，张旭光，等. 维生素E琥珀酸酯通过氧化应激诱导胃癌SGC-7901细胞凋亡的研究[J]. 癌变·畸变·突变，2010，22 (3): 186-190.

[2] 黄晓莉，赵艳，张志宏，等. 维生素E琥珀酸酯通过内质网应激诱导胃癌SGC-7901细胞凋亡的研究[J]. 癌变·畸变·突变，2012，24 (3): 195-198.

[3] Zhang X，Peng X，Yu W，et al. Alpha-tocopheryl succinate enhances doxorubicin-induced apoptosis in human gastric cancer cells via promotion of doxorubicin influx and suppression of doxorubicin efflux[J]. Cancer Lett，2011，307 (2): 174-181.

[4] Park MA，Zhang G，Martin AP，et al. Vorinostat and sorafenib increase ER stress，autophagy and apoptosis via ceramide-dependent CD95 and PERK activation[J]. Cancer Biol Ther， 2008，7 (10): 1648-1662.

[5] Li J，Yu W，Tiwary R，et al. Alpha-TEA-induced death receptor dependent apoptosis involves activation of acid sphingomyelinase and elevated ceramide-enriched cell surface membranes[J]. Cancer Cell Int，2010，10 (1): 40.

[6] Erdreich-Epstein A，Tran LB，Cox OT，et al. Endothelial apoptosis induced by inhibition of integrins alphavbeta3 and alphavbeta5 involves ceramide metabolic pathways[J]. Blood， 2005，105 (11): 4353-4361.

[7] Perrotta C，De Palma C，Falcone S，et al. Nitric oxide，ceramide and sphingomyelinase-coupled receptors: a tale of enzymes and messengers coordinating cell death，survival and differentiation [J]. Life Sci，2005，77 (14): 1732-1739.

[8] Zhu Q，Lin L，Cheng Q，et al. The role of acid sphingomyelinase and caspase 5 in hypoxia-induced HuR cleavage and subsequent apoptosis in hepatocytes[J]. Biochim Biophys Acta，2012， 1821 (12):1453-1461.

[9] Barth BM，Gustafson SJ，Hankins JL，et al. Ceramide kinase regulates TNF alpha-stimulated NADPH oxidase activity and eicosanoid biosynthesis in neuroblastoma cells[J]. Cell Signal， 2012，24 (6): 1126-1133.

[10] Lin WC，Chuang YC，Chang YS，et al. Endoplasmic reticulum stress stimulates p53 expression through NF-kappa B activation[J]. PLoS One，2012，7 (7): 39120.

[11] de Ridder G，Ray R，Misra UK，et al. Modulation of the unfolded protein response by GRP78 in prostate cancer[J]. Methods Enzymol，2011，489 (2): 245-257.

[12] Kuang E，Wan Q，Li X，et al. ER Ca2+ depletion triggers apoptotic signals for endoplasmic reticulum (ER) overload response induced by overexpressed reticulon 3 (RTN3/HAP) [J]. J Cell Physiol，2005，204 (2): 549-559.

[13] Gentile CL，Frye M，Pagliassotti MJ. Endoplasmic reticulum stress and the unfolded protein response in nonalcoholic fatty liver disease[J]. Antioxid Redox Signal，2011，15 (2): 505-521.

[14] Rojas-Rivera D，Armisén R，Colombo A，et al. TMBIM3/GRINA is a novel unfolded protein response (UPR) target gene that controls apoptosis through the modulation of ER calcium homeostasis[J]. Cell Death Differ，2012，19 (6): 1013-1026.

[15] Saito A，Ochiai K，Kondo S，et al. Endoplasmic reticulum stress response mediated by the PERK-eIF2 (alpha)-ATF4 pathway is involved in osteoblast differentiation induced by BMP2[J]. J Biol Chem，2011，286 (6): 4809-4818.

[16] Obeng EA，Boise LH. Caspase-12 and caspase-4 are not required for caspase-dependent endoplasmic reticulum stress-induced apoptosis[J]. J Biol Chem，2005，280 (33): 29578-29587.

[17] Schroder M. Endoplasmic reticulum stress responses[J]. Cell Mol Life Sci，2008，65 (6): 862-894.

[18] Oshitari T，Hata N，Yamamoto S. Endoplasmic reticulum stress and diabetic retinopathy[J]. Vasc Health Risk Manag，2008，4 (1): 115-122.

[19] Huang X，Li L，Zhang L，et al. Crosstalk between endoplasmic reticulum stress and oxidative stress in apoptosis induced by alpha-tocopheryl succinate in human gastric carcinoma cells[J]. Br J Nutr，2012，7 (1): 1-9.

Effects of acid sphingomyelinase on endoplasmic reticulum stress in VES- induced apoptosis in human gastric cancer cells

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