沉默DCR3表达增加卵巢癌细胞对紫杉醇敏感性的实验研究
摘要
目的研究沉默诱骗受体(DCR)3对卵巢癌细胞紫杉醇敏感性的影响。方法以靶向DCR3慢病毒干扰载体感染卵巢癌细胞ES-2,用Realtime PCR和Western印迹检测干扰效果。用不同浓度的紫杉醇处理ES-2细胞,细胞计数试剂盒(CCK-8)测定细胞存活情况,用紫杉醇处理沉默DCR3的ES-2细胞,CCK-8检测细胞增殖,细胞克隆实验检测细胞克隆能力,流式细胞术检测细胞凋亡,Western印迹检测细胞中活化的含半胱氨酸的天冬氨酸蛋白水解酶(C-Caspase)-12、C-Caspase-3、CCAAT/增强子结合蛋白同源蛋白(CHOP)水平。结果靶向DCR3慢病毒干扰载体下调ES-2细胞中DCR3 mRNA和蛋白水平。不同浓度的紫杉醇抑制ES-2细胞增殖,10~(-6) mol/L的紫杉醇处理后的卵巢癌细胞存活率为(53.26±4.15)%,选用10~(-6)mol/L的紫杉醇处理沉默DCR3的ES-2细胞。沉默DCR3后的ES-2细胞增殖能力显著降低,细胞克隆形成能力显著降低,细胞凋亡率显著升高,细胞中C-Caspase-12、C-Caspase-3、CHOP蛋白水平显著升高。紫杉醇处理沉默DCR3后的ES-2细胞增殖和克隆形成能力显著降低,细胞凋亡率显著升高,C-Caspase-12、C-Caspase-3、CHOP蛋白水平显著升高,与未沉默DCR3的ES-2细胞比较,差异有统计学意义(P<0.05)。结论沉默DCR3协同紫杉醇抑制卵巢癌细胞增殖,诱导卵巢癌细胞凋亡,增加卵巢癌细胞紫杉醇敏感性。
引文
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2 Takahama Y,Yamada Y,Emoto K,et al.The prognostic significance of overexpression of the decoy receptor for Fas ligand (DcR3) in patients with gastric carcinomas[J].Gastric Cancer,2002;5(2):61-8.
3 Fu Z,Chen S,Liu S,et al.DcR3 gene polymorphisms are associated with sporadic breast infiltrating ductal carcinoma in Northeast Chinese women[J].Oncotarget,2016;7(36):57970-7.
4 Sung HY,Wu HG,Ahn JH,et al.Dcr3 inhibit p53-dependent apoptosis in gamma-irradiated lung cancer cells[J].Int J Radiation Biol,2010;86(9):780-90.
5 Wu Y,Guo E,Yu J,et al.High DcR3 expression predicts stage pN2-3 in gastric cancer[J].Am J Clin Oncol,2008;54(79):79-83.
6 Xu YC,Cui J,Zhang LJ,et al.Anti-apoptosis effect of decoy receptor 3 in cholangiocarcinoma cell line TFK-1[J].Chin Med J,2018;131(1):82-7.
7 Bamias G,Kaltsa G,Siakavellas SI,et al.High intestinal and systemic levels of decoy receptor 3 (DcR3) and its ligand TL1A in active ulcerative colitis[J].Clin Immunol,2010;137(2):242-9.
8 Zhou J,Song S,He S,et al.Silencing of decoy receptor 3 (DcR3) expression by siRNA in pancreatic carcinoma cells induces Fas ligand-mediated apoptosis in vitro and in vivo[J].Int J Mol Med,2013;32(3):653-60.
9 Otsuki T,Tomokuni A,Sakaguchi H,et al.Over-expression of the decoy receptor 3 (DcR3) gene in peripheral blood mononuclear cells (PBMC) derived from silicosis patients[J].Clin Exper Immunol,2000;119(2):323-7.
10 钟智强.DcR3基因在肺癌的扩增和表达及其临床意义研究[D].福州:福建医科大学,2006.
11 陈罡.DcR3在肝癌中的表达及对肝癌细胞生长调控、迁移的影响和在凋亡中作用的研究[D].南宁:广西医科大学,2007.
12 Zhang Y,Rasul A,Li X,et al.Lentivirus-mediated shRNA targeting decoy receptor 3 (DcR3) inhibits proliferation and augments apoptosis in pancreatic cancer Capan-1 cells[J].African J Pharmacy Pharmacol,2013;7(5):203-10.
13 Simon I,Liu Y,Krall KL,et al.Evaluation of the novel serum markers B7-H4,Spondin 2,and DcR3 for diagnosis and early detection of ovarian cancer[J].Gynecol Oncol,2007;106(1):112-8.
14 Bradford L,Ziebarth A,Felder M,et al.Association of ascites and serum decoy receptor 3 (DCR3) levels with tumor infiltrating lymphocytes (TILS) and tumor vessels counts in epithelial ovarian cancer (EOC)[J].Gynecol Oncol,2010;116(3):598.
15 乔子晗.紫杉醇抗肿瘤机制和临床应用研究[J].临床医药文献电子杂志,2016;3(40):8085-6.
16 徐晚成.抗癌药物紫杉醇的作用机理及机制[J].滨州医学院学报,2017;40(4):294-7.
17 Takei Y,Kadomatsu K,Goto T,et al.Combinational antitumor effect of siRNA against midkine and paclitaxel on growth of human prostate cancer xenografts[J].Cancer,2006;107(4):864-73.
18 Wei W,Lv PP,Chen XM,et al.Codelivery of mTERT siRNA and paclitaxel by chitosan-based nanoparticles promoted synergistic tumor suppression[J].Biomaterials,2013;34(15):3912-23.
19 Liu Y,Wang Y,Sun X,et al.miR-449a promotes liver cancer cell apoptosis by downregulation of calpain 6 and POU2F1[J].Oncotarget,2016;7(12):13491-501.
20 Su CC.Tanshinone ⅡA could inhibit pancreatic cancer BxPC-3 cells through increasing PERK,ATF6,caspase-12 and CHOP expression to induce apoptosis[J].J Biomedical Sci Engineer,2015;8(3):149-59.
21 Liu G,Su L,Hao X,et al.Salermide up-regulates death receptor 5 expression through the ATF4-ATF3-CHOP axis and leads to apoptosis in human cancer cells[J].J Cell Mol Med,2012;16(7):1618-28.
22 Lee JY,Jung KH,Morgan MJ,et al.Sensitization of TRAIL-induced cell death by 20(S)-ginsenoside Rg3 via CHOP-mediated DR5 upregulation in human hepatocellular carcinoma cells[J].Mol Cancer Therapeutics,2013;12(3):274-85.
23 Yan MY,Chien SY,Kuo SJ,et al.Tanshinone ⅡA inhibits BT-20 human breast cancer cell proliferation through increasing caspase 12,GADD153 and phospho-p38 protein expression[J].Int J Mol Med,2012;29(5):855-63.
24 Wang W,Yan H,Dou C,et al.Human leptin triggers proliferation of A549 cells via blocking endoplasmic reticulum stress-related apoptosis[J].Biochem Biokhimiia,2013;78(12):1333-41.
25 Huang Y,Li X,Wang Y,et al.Endoplasmic reticulum stress-induced hepatic stellate cell apoptosis through calcium-mediated JNK/P38 MAPK and Calpain/Caspase-12 pathways[J].Mol Cell Biochem,2014;394(1-2):1-12.