• 1.解放軍第451醫(yī)院肝膽普外科(陜西西安 710054);;
  • 2.第四軍醫(yī)大學(xué)西京醫(yī)院消化病研究所(陜西西安 710032);;
  • 3.第四軍醫(yī)大學(xué)西京醫(yī)院肝膽外科(陜西西安 710032);

目的探討腫瘤相關(guān)糖蛋白72(TAG72)靶向性嵌合錨定T淋巴細(xì)胞的制備方法，并檢測(cè)它對(duì)TAG72陽(yáng)性肝癌細(xì)胞增殖的阻滯效應(yīng)。方法分離外周血單核細(xì)胞(PBMC)，然后用免疫磁珠法分離得到CD8+ T淋巴細(xì)胞。將重組真核表達(dá)載體anti-TAG72-scFv-CD28-pcDNA3.0采用脂質(zhì)體介導(dǎo)的細(xì)胞轉(zhuǎn)染和細(xì)胞培養(yǎng)，以制備TAG72靶向性的嵌合錨定T淋巴細(xì)胞; 將嵌合錨定T淋巴細(xì)胞與TAG72陽(yáng)性肝癌細(xì)胞SMMC7721共培養(yǎng)，通過(guò)流式細(xì)胞儀檢測(cè)肝癌細(xì)胞的周期變化，分析嵌合錨定T淋巴細(xì)胞對(duì)肝癌細(xì)胞增殖的抑制效應(yīng)。結(jié)果TAG72靶向性嵌合錨定T淋巴細(xì)胞可識(shí)別肝癌細(xì)胞SMMC7721； 用流式細(xì)胞儀檢測(cè)發(fā)現(xiàn)，嵌合錨定T淋巴細(xì)胞可引起肝癌細(xì)胞SMMC7721的增殖阻滯。結(jié)論TAG72靶向性嵌合錨定T淋巴細(xì)胞可特異性識(shí)別TAG72陽(yáng)性肝癌細(xì)胞SMMC7721并引起其增殖阻滯。

引用本文： 徐宏勇,徐立,李開(kāi)宗. 嵌合錨定T淋巴細(xì)胞對(duì)肝癌細(xì)胞增殖的阻滯效應(yīng). 中國(guó)普外基礎(chǔ)與臨床雜志, 2011, 18(4): 370-373. doi: 復(fù)制

1. 徐宏勇, 徐立, 李開(kāi)宗, 等. 抗TAG72抗原單鏈抗體的原核表達(dá)及對(duì)人肝癌的檢測(cè) [J]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2010, 17(7): 699702.
2. Ponnusamy MP, Venkatraman G, Singh AP, et al. Expression of TAG72 in ovarian cancer and its correlation with tumor stage and patient prognosis [J]. Cancer Lett，2007, 251(2): 247257.
3. Chauhan SC, Vinayek N, Maher DM, et al. Combined staining of TAG72, MUC1, and CA125 improves labeling sensitivity in ovarian cancer: antigens for multitargeted antibodyguided therapy [J]. J Histochem Cytochem, 2007, 55(8): 867875.
4. June CH, Bluestone JA, Nadler LM, et al. The B7 and CD28 receptor families [J]. Immunol Today, 1994, 15(7): 321331.
5. Petersen CC, Petersen MS, Agger R, et al. Accumulation in tumor tissue of adoptively transferred T cells: A comparison between intravenous and intraperitoneal injection [J]. J Immunother, 2006, 29(3): 241249.
6. 徐宏勇, 徐立, 李開(kāi)宗, 等. 靶向性腫瘤相關(guān)抗原TAG72的scFvCD28嵌合基因的真核表達(dá)載體的構(gòu)建 [J].中國(guó)普外基礎(chǔ)與臨床雜志, 2002, 9(6): 398401.
7. 徐宏勇, 徐立, 高建宏, 等. 嵌合錨定T細(xì)胞致癌胚抗原陽(yáng)性胃癌細(xì)胞的凋亡效應(yīng) [J]. 中華醫(yī)學(xué)雜志, 2007, 87(15): 10531057.
8. Willemsen RA, Weijtens ME, Ronteltap C, et al. Grafting primary human T lymphocytes with cancerspecific chimeric single chain and two chain TCR [J]. Gene Ther, 2000, 7(16): 13691377.
9. Ychou M, Duffour J, Kramar A, et al. Clinical significance and prognostic value of CA724 compared with TAG72 and CA199 in patients with gastric cancer [J]. Dis Markers, 2000, 16(34): 105110.
10. Hombach A, Heuser C, Marquardt T, et al. CD4+ T cells engrafted with a recombination immunorecrpter efficiently lyse target cells in a MHC antigen and Fas independent fashion [J]. J Immuno, 2001, 167(2): 10901095.
11. Hombach A, Wieczarkowiecz A, Marquardt T, et al. Tumorspecific T cell activation by recombinant immunoreceptors: CD3 zeta signaling and CD28 costimulation are simultaneously required for efficient IL2 secretion and can be integrated into one combined CD28/CD3 zeta signaling receptor molecule [J]. J Immunol, 2001, 167(11): 61236130.
12. Lee EY, Kim C, Kim MJ, et al. Signet ring cell carcinoma of the extrahepatic bile duct [J]. Gut Liver, 2010, 4(3): 402406.
13. Gupta S, Sathishkumar S, Ahmed MM. influence of cell cycle checkpoints and p53 function on the toxicity of temozolomide in human pancreatic cancer cells [J]. Pancreatology, 2010, 10(5): 565579.
14. Spitzner M, Emons G, Kramer F, et al. A gene expression signature for chemoradiosensitivity of colorectal cancer cells [J]. Int J Radiat Oncol Biol Phys, 2010, 78(4): 11841192.
15. WilliamFaltaos S, Rouillard D, Lechat P, et al. Cell cycle arrest and apoptosis induced by oxaliplatin (LOHP) on four human cancer cell lines [J]. Anticancer Res, 2006, 26(3A): 20932099.
16. WilliamFaltaos S, Rouillard D, Lechat P, et al. Cell cycle arrest by oxaliplatin on cancer cells [J]. Fundam Clin Pharmacol, 2007, 21(2): 165172.
  1. 1. 徐宏勇, 徐立, 李開(kāi)宗, 等. 抗TAG72抗原單鏈抗體的原核表達(dá)及對(duì)人肝癌的檢測(cè) [J]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2010, 17(7): 699702.
  2. 2. Ponnusamy MP, Venkatraman G, Singh AP, et al. Expression of TAG72 in ovarian cancer and its correlation with tumor stage and patient prognosis [J]. Cancer Lett，2007, 251(2): 247257.
  3. 3. Chauhan SC, Vinayek N, Maher DM, et al. Combined staining of TAG72, MUC1, and CA125 improves labeling sensitivity in ovarian cancer: antigens for multitargeted antibodyguided therapy [J]. J Histochem Cytochem, 2007, 55(8): 867875.
  4. 4. June CH, Bluestone JA, Nadler LM, et al. The B7 and CD28 receptor families [J]. Immunol Today, 1994, 15(7): 321331.
  5. 5. Petersen CC, Petersen MS, Agger R, et al. Accumulation in tumor tissue of adoptively transferred T cells: A comparison between intravenous and intraperitoneal injection [J]. J Immunother, 2006, 29(3): 241249.
  6. 6. 徐宏勇, 徐立, 李開(kāi)宗, 等. 靶向性腫瘤相關(guān)抗原TAG72的scFvCD28嵌合基因的真核表達(dá)載體的構(gòu)建 [J].中國(guó)普外基礎(chǔ)與臨床雜志, 2002, 9(6): 398401.
  7. 7. 徐宏勇, 徐立, 高建宏, 等. 嵌合錨定T細(xì)胞致癌胚抗原陽(yáng)性胃癌細(xì)胞的凋亡效應(yīng) [J]. 中華醫(yī)學(xué)雜志, 2007, 87(15): 10531057.
  8. 8. Willemsen RA, Weijtens ME, Ronteltap C, et al. Grafting primary human T lymphocytes with cancerspecific chimeric single chain and two chain TCR [J]. Gene Ther, 2000, 7(16): 13691377.
  9. 9. Ychou M, Duffour J, Kramar A, et al. Clinical significance and prognostic value of CA724 compared with TAG72 and CA199 in patients with gastric cancer [J]. Dis Markers, 2000, 16(34): 105110.
  10. 10. Hombach A, Heuser C, Marquardt T, et al. CD4+ T cells engrafted with a recombination immunorecrpter efficiently lyse target cells in a MHC antigen and Fas independent fashion [J]. J Immuno, 2001, 167(2): 10901095.
  11. 11. Hombach A, Wieczarkowiecz A, Marquardt T, et al. Tumorspecific T cell activation by recombinant immunoreceptors: CD3 zeta signaling and CD28 costimulation are simultaneously required for efficient IL2 secretion and can be integrated into one combined CD28/CD3 zeta signaling receptor molecule [J]. J Immunol, 2001, 167(11): 61236130.
  12. 12. Lee EY, Kim C, Kim MJ, et al. Signet ring cell carcinoma of the extrahepatic bile duct [J]. Gut Liver, 2010, 4(3): 402406.
  13. 13. Gupta S, Sathishkumar S, Ahmed MM. influence of cell cycle checkpoints and p53 function on the toxicity of temozolomide in human pancreatic cancer cells [J]. Pancreatology, 2010, 10(5): 565579.
  14. 14. Spitzner M, Emons G, Kramer F, et al. A gene expression signature for chemoradiosensitivity of colorectal cancer cells [J]. Int J Radiat Oncol Biol Phys, 2010, 78(4): 11841192.
  15. 15. WilliamFaltaos S, Rouillard D, Lechat P, et al. Cell cycle arrest and apoptosis induced by oxaliplatin (LOHP) on four human cancer cell lines [J]. Anticancer Res, 2006, 26(3A): 20932099.
  16. 16. WilliamFaltaos S, Rouillard D, Lechat P, et al. Cell cycle arrest by oxaliplatin on cancer cells [J]. Fundam Clin Pharmacol, 2007, 21(2): 165172.