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[目的]利用已构建好的肿瘤抑素相关活性肽T42肽高效表达基因工程重组菌pTYB2-T42提取纯化T42肽,验证T42肽联合环磷酰胺后抗肿瘤活性。[方法]经几丁质柱亲和层析目的蛋白纯化,Tricine-SDS-PAGE分析后用紫外分光光度法测其浓度,进行体外肿瘤细胞MTT实验,利用流式细胞仪检测肿瘤细胞凋亡。[结果]T42肽单独及与环磷酰胺联合用药均使人肝癌Hepg2细胞生长增殖受到抑制,其IC50分别为30μmol/ml、25μmol/ml,呈剂量依赖,而PBS组和空白组无明显变化,且两者均能促Hepg2细胞凋亡;联合环磷酰胺后促凋亡作用更加明显。[结论]该研究结果将为肿瘤抑素抗肿瘤的临床治疗提供重要的实验基础,为寻找既能增强抗肿瘤作用又能降低放疗、化疗药物毒副作用的联合用药配伍方式提供借鉴。
[Objective] The purpose of this study was to validate the anti-tumor activity of T42 peptide combined with cyclophosphamide after the constructed T42 tumor-related peptide T42 peptide was highly expressed in pTYB2-T42. [Method] Purification of target protein by chitin affinity chromatography, Tricine-SDS-PAGE analysis and UV spectrophotometry to determine the concentration of tumor cells in vitro MTT assay, the use of flow cytometry to detect apoptosis of tumor cells. [Results] T42 peptide alone and combined with cyclophosphamide both inhibited the growth of human HepG2 HepG2 cells. The IC50 values were 30 μmol / ml and 25 μmol / ml, respectively, in a dose-dependent manner. No significant changes were observed in PBS group and blank group, Both of them can promote the apoptosis of Hepg2 cells. Combined with cyclophosphamide, the pro-apoptotic effect is more obvious. [Conclusion] The results of this study will provide an important experimental basis for the clinical treatment of tumor suppressor tumor, and provide a reference for finding out the combination modality that can both enhance the anti-tumor effect and reduce the side effects of radiotherapy and chemotherapy drugs.