中国肿瘤生物治疗杂志2011，Vol.18Issue(6)：597-604,8.DOI:10.3872/j.issn.1007-385X.2011.06.003
缺氧微环境对TSST-1诱导的抗CEA+结肠癌LoVo细胞免疫治疗的调控
Hypoxia microenvironment regulates immunotherapy effect of TSST-1 on CEA positive colon cancer LoVo cells
摘要
Abstract
Objective: To evaluate the regulation of hypoxia microenvironment on cytotoxicity effect of peripheral blood lymphocyte (PBL) activated by the superantigen of toxic shock syndrome toxin-1 ( TSST-1 ) against CEA+ human colon cancer cell line LoVo. Methods: The recombinant retroviral vector pLEGFP-Nl-5HRE-CEAp-TSST-l-linker-CD80TM (pLEGFP-Nl-5HCTC) containing a CEA promoter and 5 copies of the hypoxia-response elements (HRE) enhancer, which was constructed in our previous study, was packaged and collected. pLEGFP-Nl-5HCTC was then transducted into CEA + LoVo cells or CEA ~ human cervical carcinoma HeLa cells, and LoVo or HeLa cells with stable expression of trans-membrane superantigen fusion gene TSST-1 -linker-CD80TM protein (TC fusion protein) were obtained. Hypoxia microenvironment was simulated by CoCl2, and RT-PCR and Western blotting were employed to examine the expression of TSST-1 regulated by hypoxia. Human PBL was extracted and co-cultured with transgene tumor cells infected with pLEGFP-Nl-5HCTC retrovirus; H-TdR assay was employed to detect the proliferation of PBL regulated by hypoxia; and the cytotocixi-ty effect of PBL regulated by hypoxia against the transgene tumor cells was detected by MTT assay. Results; CEA + LoVo cells infected with recombinant retrovirus pLEGFP-Nl-5HCTC were obtained (5 HCTC/LoVo). The expression of TSST-1 mRNA and protein in 5HCTC/LoVo cells was further increased with hypoxia, which was confirmed by RT-PCR and Western blotting. There was no TSST-1 expression in CEA ~ HeLa cells under either hypoxic condition or normoxic condition. The proliferation of PBL activated by 5HCTC/LoVo cells was increased with hypoxia (7. 3 × 103 vs 3. 1 × 103 cpm, P < 0. 05) ; and the cytotocixity of PBL on 5HCTC/LoVo cells was more effective in hypoxia condition than in normoxic condition (82. 69% vs 53. 50% , P <0. 01) . There was no proliferation of PBL activated by CEA" HeLa cells, and PBL also could not inhibit the proliferation of CEA" HeLa cells. Conclusion: Hypoxia microenvironment can increase the cytotocixity effect of targeting expressed superantigen TSST-1 on CEA + LoVo cells.关键词
缺氧微环境/中毒性休克综合征毒素-1(TSST-1)/CEA+结肠癌细胞/LoVo细胞/HeLa细胞/逆转录病毒/外周血淋巴细胞Key words
hypoxia environment/ toxic shock syndrome toxin-1 (TSST-1) / CEA positive tumors cells/ LoVo cell/ HeLa cell/ retrovirus/ peripheral blood lymphocyte分类
医药卫生引用本文复制引用
王炜,孙学军,王伟,田勇,郑见宝,成亮,张超..缺氧微环境对TSST-1诱导的抗CEA+结肠癌LoVo细胞免疫治疗的调控[J].中国肿瘤生物治疗杂志,2011,18(6):597-604,8.基金项目
国家自然科学基金资助项目(No.30672070,No.30400430) （No.30672070,No.30400430）
陕西省科学技术研究发展计划资助项目(No.2009 K01-70). （No.2009 K01-70）
