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人肿瘤坏死因子相关凋亡诱导配体(hTRAIL)因对肿瘤细胞有选择性杀伤作用而成为新型抗肿瘤候选药物。预测的恒河猴TRAIL(mmTRAIL)与hTRAIL高度同源,提示其有杀伤人肿瘤细胞的可能性。但目前尚无mmTRAIL蛋白功能的报道。本文首先从恒河猴外周血单个核淋巴细胞中扩增了可溶性mmTRAIL编码基因,然后构建了pQE30-mmTRAIL表达质粒并进行诱导表达。通过Ni-NTA agarose亲和层析从破菌上清液中获得mmTRAIL蛋白。聚丙烯酰胺凝胶电泳和凝胶过滤层析表明,mmTRAIL在溶液中主要以三聚体形式存在。体外条件下,mmTRAIL对结肠癌细胞COLO205显示强烈的杀伤作用,而对正常细胞BEA2b无害,表明其对肿瘤细胞的杀伤有选择性。裸鼠荷瘤模型进一步证实mmTRAIL能够抑制肿瘤生长,具有抗肿瘤作用。这些结果表明,mmTRAIL能够杀伤人肿瘤细胞,可作为新型肿瘤靶向治疗药物进一步深入研究。
Human tumor necrosis factor-related apoptosis-inducing ligand (hTRAIL) has become a new antitumor drug candidate because of its selective killing effect on tumor cells. The predicted rhesus TRAIL (mmTRAIL) is highly homologous to hTRAIL, suggesting its potential for killing human tumor cells. However, there is no report about the function of mmTRAIL protein. In this paper, the soluble mmTRAIL gene was amplified from peripheral blood mononuclear lymphocytes of rhesus monkeys. Then, the expression plasmid pQE30-mmTRAIL was constructed and induced. The mmTRAIL protein was obtained from the broken bacteria supernatant by Ni-NTA agarose affinity chromatography. Polyacrylamide gel electrophoresis and gel filtration chromatography showed that mmTRAIL is predominantly in the form of a trimer in solution. In vitro, mmTRAIL showed a strong killing effect on colon cancer cells COLO205, but not on normal cells BEA2b, indicating its selective killing of tumor cells. Nude mice tumor model further confirmed that mmTRAIL can inhibit tumor growth, with anti-tumor effect. These results indicate that mmTRAIL can kill human tumor cells and can be further studied as a novel tumor-targeted therapeutic.