• 四川大學(xué)華西第二醫(yī)院（成都，610041）1婦產(chǎn)科，2病理科;

【摘要】　目的　 研究腫瘤壞死因子相關(guān)凋亡誘導(dǎo)配體（TRAIL）蛋白對SKOV3移植瘤細胞半胱天冬氨酸蛋白酶-3（Caspase-3）表達的影響及其與腫瘤細胞凋亡的關(guān)系。　方法　 建立雌性裸小鼠SKOV3移植瘤24只，隨機分為4組，每組6只。TRAIL組單用重組人TRAIL蛋白（10 μg/kg），順鉑（DDP）組單用DDP（3 mg/kg），TRAIL+DDP組聯(lián)合使用TRAIL蛋白（10 μg/kg）和DDP（3 mg/kg），空白對照組給予0.5 mL生理鹽水。經(jīng)處理后，各組的組織切片用免疫組織化學(xué)染色檢測Caspase-3的表達和末端脫氧核苷酸轉(zhuǎn)移酶介導(dǎo)核苷酸缺口標(biāo)記技術(shù)（TUNEL）檢測腫瘤細胞凋亡指數(shù)。　結(jié)果　 Caspase-3的表達水平在TRAIL組（171.67±14.38）、DDP組（172.50±14.75）、聯(lián)合組（230.00±40.99）中均明顯高于對照組（135.83±16.25）（P lt;0.05）。SKOV3移植瘤細胞凋亡指數(shù)在空白對照組、TRAIL組、DDP組和聯(lián)合組分別為16.67±5.43、33.17±8.42、24.33±4.59和40.50±6.16，TRAIL組和聯(lián)合組細胞凋亡指數(shù)較空白對照組和DDP組明顯增高（P lt;0.05）。　結(jié)論　 TRAIL蛋白使卵巢癌移植瘤細胞的Caspase-3表達增強，TRAIL蛋白促進腫瘤細胞凋亡發(fā)生。
【Abstract】　Objective　 To investigate the effects of Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) on the expression of Cysteine/aspartic acid specific protease- 3 (Caspase-3) in SKOV3 ovarian tumor cells and its relationship with the apoptosis of the ovarian tumor xenografts in nude mice. 　Methods　 Twenty-four nude mice with SKOV3 cell line ovarian tumor were randomly divided into four groups with 6 in each group. TRAIL (10 μg/kg) was given to the mice in the TRAIL group; DDP (3 μg/kg) was given to the mice in the DDP group; TRAIL (10 μg/kg) and DDP (3 μg/kg) were given to the mice in the TRAIL+DDP group; and 0.5 mL of saline solution was give to the mice in the control group. The expression of Caspase-3 was detected with immunohistochemistry. The apoptosis index (AI) of cells was determined by Terminal deoxynucleotidyl transferase mediated-dUTP nick end labeling (TUNEL).　Results　 The expression of Caspase-3 in the TRAIL group (171.67±14.38), DDP group (172.50±14.75), and TRAIL+DDP group (230.00±40.99) was significantly higher than that in the control group (135.83±16.25) (P lt;0.05). The apoptosis index for the control group, TRAIL group, DDP group and TRAIL+DDP group was 16.67±5.43, 33.17±8.42, 24.33±4.59, and 40.50±6.16, respectively. The apoptosis index for the TRAIL group and the TRAIL+DDP group was significantly higher than that in the control group and the DDP group (P lt;0.05).　Conclusion　 Soluble TRAIL has an effect on enhancing the expression of Caspase-3 in implanted tumor in nude mice. TRAIL protein can inhibit the growth of SKOV3 cells in nude mice by inducing cell apoptosis.

引用本文： 李春梅,彭芝蘭,楊開選,劉英,黃琴. 腫瘤壞死因子相關(guān)凋亡誘導(dǎo)配體對卵巢癌裸鼠移植瘤SKOV3細胞的促凋亡研究. 華西醫(yī)學(xué), 2011, 26(4): 481-484. doi: 復(fù)制

1. 　Cretney E, Takeda K, Yagita H, et al. Increased susceptibility to tumor initiation and metastasis in TNF-related apoptosis-inducing ligand-deficient mice[J]. J Immunol, 2002, 168(3): 1356-1361.
2. 　MeKay TR, Bell S, Tenev T, et al. Procaspase 3 expression in ovarian carcinoma cells increases survivin transcription which can be countered with a dominant-negative mutant, survivin T34A; a combination gene therapy strategy[J].Oncogene, 2003, 22(23): 3539-3547.
3. 　李春梅, 彭芝蘭, 尹如鐵, 等. TRAIL對人卵巢癌SKOV3細胞生長及凋亡的影響[J]. 四川大學(xué)學(xué)報(醫(yī)學(xué)版), 2006, 37(5): 761-764.
4. 　Kischkel FC, Lawrence DA, Chuntharapai A, et al. Apo2L/ TRAIL-dependent recruitment of endogenous FADD and caspase-8 to death receptors 4 and 5[J]. Immunity, 2000, 12(6): 611-620.
5. 　Lin ML, Lu YC, Chung JG, et al. Aloe-emodin induces apoptosis of human nasopharyngeal carcinoma cells via caspase-8-mediated activation of the mitochondrial death pathway[J]. Cancer Lett, 2010, 291(1): 46-58.
6. 　Jin H, Yang R, Fong S, et al. Apo2 ligand/tumor necrosis factor-related apoptosis-inducing ligand cooperates with chemotherapy to inhibit orthotopic lung tumor growth and improve survival[J]. Cancer Res, 2004, 64(14): 4900-4905.
7. 　Shankar S, Singh TR, Chen X, et al. The sequential treatment with ionizing radiation followed by TRAIL/Apo-2L reduces tumor growth and induces apoptosis of breast tumor xenografts in nude mice[J]. Int J Oncol, 2004, 24(5): 1133-1140.
8. 　Bellail AC, Qi L, Mulligan P, et al. TRAIL agonists on clinical trials for cancer therapy: the promises and the challenges[J]. Rev Recent Clin Trials, 2009, 4(1): 34-41.
9. 　Takeda K, Yamaguchi N, Akiba H, et al. Induction of tumor-specific T cell immunity by anti-dr5 antibody therapy[J]. J Exp Med, 2004, 199(4): 437-448.
10. 　Johnstone RW, Frew AJ, Smyth MJ. The TRAIL apoptotic pathway in cancer onset, progression and therapy[J]. Nat Rev Cancer, 2008, 8(10): 782-798.
11. 　El-Gazzar A, Perco P, Eckelhart E, et al. Natural immunity enhances the activity of a DR5 agonistic antibody and carboplatin in the treatment of ovarian cancer[J]. Mol Cancer Ther, 2010, 9(4): 1007-1018.
12. 　李春梅, 彭芝蘭, 尹如鐵. TRAIL蛋白與順鉑聯(lián)合應(yīng)用對卵巢癌裸鼠腹腔移植瘤的生長抑制作用[J]. 中華婦產(chǎn)科雜志, 2006, 41(8): 568-556.
13. 　Gliniak B, Le T. Tumor necrosis factor-related apoptosis-inducing ligand’s antitumor activity in vivo is enhanced by the chemotherapeutic agent CPT-11[J]. Cancer Res, 1999, 59(24): 6153-6158.
14. 　Nagane M, Pan G, Weddle J, et al. Increased death receptor 5 expression by chemotherapeutic agents in human gliomas causes synergistic cytotoxicity with tumor necrosis factor-related apoptosis-inducing ligand in vitro and in vivo[J]. Cancer Res, 2000, 60(4): 847-853.
  1. 1. 　Cretney E, Takeda K, Yagita H, et al. Increased susceptibility to tumor initiation and metastasis in TNF-related apoptosis-inducing ligand-deficient mice[J]. J Immunol, 2002, 168(3): 1356-1361.
  2. 2. 　MeKay TR, Bell S, Tenev T, et al. Procaspase 3 expression in ovarian carcinoma cells increases survivin transcription which can be countered with a dominant-negative mutant, survivin T34A; a combination gene therapy strategy[J].Oncogene, 2003, 22(23): 3539-3547.
  3. 3. 　李春梅, 彭芝蘭, 尹如鐵, 等. TRAIL對人卵巢癌SKOV3細胞生長及凋亡的影響[J]. 四川大學(xué)學(xué)報(醫(yī)學(xué)版), 2006, 37(5): 761-764.
  4. 4. 　Kischkel FC, Lawrence DA, Chuntharapai A, et al. Apo2L/ TRAIL-dependent recruitment of endogenous FADD and caspase-8 to death receptors 4 and 5[J]. Immunity, 2000, 12(6): 611-620.
  5. 5. 　Lin ML, Lu YC, Chung JG, et al. Aloe-emodin induces apoptosis of human nasopharyngeal carcinoma cells via caspase-8-mediated activation of the mitochondrial death pathway[J]. Cancer Lett, 2010, 291(1): 46-58.
  6. 6. 　Jin H, Yang R, Fong S, et al. Apo2 ligand/tumor necrosis factor-related apoptosis-inducing ligand cooperates with chemotherapy to inhibit orthotopic lung tumor growth and improve survival[J]. Cancer Res, 2004, 64(14): 4900-4905.
  7. 7. 　Shankar S, Singh TR, Chen X, et al. The sequential treatment with ionizing radiation followed by TRAIL/Apo-2L reduces tumor growth and induces apoptosis of breast tumor xenografts in nude mice[J]. Int J Oncol, 2004, 24(5): 1133-1140.
  8. 8. 　Bellail AC, Qi L, Mulligan P, et al. TRAIL agonists on clinical trials for cancer therapy: the promises and the challenges[J]. Rev Recent Clin Trials, 2009, 4(1): 34-41.
  9. 9. 　Takeda K, Yamaguchi N, Akiba H, et al. Induction of tumor-specific T cell immunity by anti-dr5 antibody therapy[J]. J Exp Med, 2004, 199(4): 437-448.
  10. 10. 　Johnstone RW, Frew AJ, Smyth MJ. The TRAIL apoptotic pathway in cancer onset, progression and therapy[J]. Nat Rev Cancer, 2008, 8(10): 782-798.
  11. 11. 　El-Gazzar A, Perco P, Eckelhart E, et al. Natural immunity enhances the activity of a DR5 agonistic antibody and carboplatin in the treatment of ovarian cancer[J]. Mol Cancer Ther, 2010, 9(4): 1007-1018.
  12. 12. 　李春梅, 彭芝蘭, 尹如鐵. TRAIL蛋白與順鉑聯(lián)合應(yīng)用對卵巢癌裸鼠腹腔移植瘤的生長抑制作用[J]. 中華婦產(chǎn)科雜志, 2006, 41(8): 568-556.
  13. 13. 　Gliniak B, Le T. Tumor necrosis factor-related apoptosis-inducing ligand’s antitumor activity in vivo is enhanced by the chemotherapeutic agent CPT-11[J]. Cancer Res, 1999, 59(24): 6153-6158.
  14. 14. 　Nagane M, Pan G, Weddle J, et al. Increased death receptor 5 expression by chemotherapeutic agents in human gliomas causes synergistic cytotoxicity with tumor necrosis factor-related apoptosis-inducing ligand in vitro and in vivo[J]. Cancer Res, 2000, 60(4): 847-853.