用不同体外实验评价了转基因大米Bt63的细胞毒性和遗传毒性。分别以200、100、50、25、12.5 μg/mL转基因大米Bt63全蛋白作用于人淋巴细胞，并各培养1.5、6、24 h。然后用彗星试验检测DNA损伤，用微核试验检测染色体畸变，用CCK8及中性红试验检测细胞毒性损伤。在经过不同的培养时间后，阳性对照组淋巴细胞的遗传及细胞损伤指标与对照组相比存在显著差异。其中，CCK8试验、中性红试验测得细胞存活率存在着明显的损伤作用时间效应关系。转基因大米Bt63全蛋白暴露组淋巴细胞遗传及细胞损伤指标与非转基因大米全蛋白暴露组相比无明显差异，且与对照差异不显著。转基因大米Bt63与非转基因大米两者在遗传毒性及细胞毒性上性状相似，实质等同。转基因大米Bt63对人淋巴细胞无明显遗传毒性及细胞毒性作用。另外，该实验体系与传统的毒理学方法相比，经济、简便、快速，具有较高的实用价值。
陈吴健1,郑伟2,*,吕沁风2,张明哲1,吴忠华2,徐琦2,吴志毅1,李禾2
. 体外试验评价转基因大米Bt63的安全性[J]. 中国水稻科学, 2012
, 26(5)
: 633
-636
.
DOI: 10.3969/j.issn.10017216.2012.05.019
To investigate the cytogenotoxicity of Bt63, a genetically modified(GM) rice in human lymphocytes in vitro, human lymphocytes were exposed to whole protein of Bt63 at doses of 200, 100, 50, 25, 12.5 μg/mL and incubated for 1.5 h, 6 h and 24 h, respectively. The DNA damage, chromosomal aberrations and cytotoxicity induced by whole protein of Bt63 were measured by comet assay, micronucleus test, CCK8 assay and neutral red uptake (NRU) assay. After different incubation periods, the cytogenotoxicity of lymphocytes in positive control group was significant higher than that of lymphocytes in blank control group. Moreover, the exposure timeeffect relationship was observed in positive control group by CCK8 assay and neutral red uptake (NRU) assay. There was no significant difference in cytogenotoxicity between Bt63 group and nonGM rice group. Also, the Bt63 GM rice group did not show higher cytogenotoxicity than that of nonGM rice group, indicating that Bt63 GM rice has similar cytogenotoxicity with nonGM rice, and Bt63 GM rice has no cytogenotoxicity effect on human lymphocytes in vitro. Compared with traditional toxicological methods, the experimental system is economic, simple, rapid and reliable.
\[1\]Cummins J, Ho M W, Ryan A. Hazardous CaMV promoter? Nat Biotechnol, 2000, 18: 363.
\[2\]Key S, Ma J K, Drake P M. Genetically modified plants and human health. J R Soc Med, 2008, 101(6): 290298.
\[3\]顾祖维. 转基因食品的安全性及其毒理学评价. 毒理学杂志, 2005, 19(1): 911.
\[4\]焦哲, 李攻科. 转基因食品及其安全性评价. 大学化学, 2007, 22(4): 3137.
\[5\]张文平, 李云云, 王伟刚, 等. 转几丁质酶和β1， 3葡聚糖基因玉米的致突变性检测. 中国药物与临床, 2009, 9(5): 405407.
\[6\]Anldanl E, Gadani F, Heinze P. Analytical methods for detection and determination of genetically modified organisms in agricultural crops and plantderived food products. Eur Food Res Technol, 2002, 214(1): 326.
\[7\]Pereira A. A transgenic perspective on plant functional genomics. Transgenic Res, 2000, 9(415): 245260.
\[8\]Millstone E, Brunner E, Mayer S. Beyond substantial equivalence. Nature, 1999, 401(7): 525526.
\[9\]Kuiper H A, Noteborn H P, Peijnenbury A A. Adequacy of methods for testing the safety of genetically modified foods. Lancet, 1999, 354: 13151316.
\[10\]Bryan D, James D A, Helen C, et al. Evaluation of protein safety in the context of agricultural biotechnology. Food Chem Toxicol, 2008, (46): 7197.
\[11\]Kristina Hug. Genetically modified organisms: Do the benefits outweigh the risks? Medicina (Kaunas), 2008, 44(2): 8799.
\[12\]刘岭峰, 曹江山, 吴文花, 等. 转基因食品安全性评价及我国管理现状. 江西农业学报, 2009, 21(4): 155158.
\[13\]Kuipera H A, Konig A, Kletera G A, et al. Concluding remarks. Food Chem Toxicol, 2004, 42: 11951202.
\[14\]Anthony J C, Jeanne M E J. Genetic engineering of crops as potential source of genetic hazard in the human diet. Mut Res, 1999, 443: 223234.
\[15\]Konig A, Cockburnb A, Crevelc R W R et al. Assessment of the safety of foods derived from genetically modified (GM) crops. Food Chem Toxicol, 2004, (42): 10471088.
\[16\]Kok E J, Keijer J, Kleter G A, et al. Comparative safety assessment of plantderived foods. Regul Toxicol Pharmacol, 2008, 50: 98113.
