中文摘要：黑色素瘤是目前唯一一种致命的常见恶性皮肤肿瘤，很容易恶化和转移。本研究评价了牛蛙皮肤中改性颞叶蛋白-la (T-La)肽活性的抗肿瘤活性，并提高了其在黑色素瘤细胞中的靶向性。修饰T-La的氨基酸序列，得到抗肿瘤肽T-La (FS)；将T-La和T-La (FS)分别与RGD小分子多肽偶联，形成嵌合肽RGD-T-La和RGD-T-La (FS)。圆二色法评估发现嵌合肽的二级结构为α螺旋。利用生物信息学研究了T-La的结构。此外，还分析了修饰肽的抗肿瘤作用以及RGD嵌合肽对肿瘤的体内外靶向作用。采用MTT法测定体外抗肿瘤活性；利用流式细胞术检测高表达整合素αvβ3的肿瘤细胞；筛选肿瘤细胞对RGD-T-La (FS)的敏感性，建立裸鼠肿瘤模型；利用激光共聚焦显微镜实时测量几种肽对肿瘤细胞的影响；用扫描电镜观察其抗肿瘤作用机制。研究发现，B16黑色素瘤细胞对肽最敏感，10 mu g/ml RGD-T-La (FS)的细胞存活率为24.65%；RGD-La (FS)对肿瘤细胞起效迅速；RGD嵌合肽在肿瘤细胞中具有靶向性杀伤作用；在B16黑色素瘤小鼠模型中，这些肽可能通过抑制VEGF和促进caspase-3的表达，抑制黑色素瘤早期发展，诱导肿瘤凋亡，具有抗肿瘤作用。总之，本研究为小分子抗菌肽作为靶向抗肿瘤药物的应用提供了科学依据，为其作为抗肿瘤药物的临床应用奠定了基础。
外文摘要：Melanoma is a common malignant skin tumor, which is the only fatal skin tumor at present. Melanoma has a high degree of malignancy and metastasis. The activity of modified Temporin-La (T-La) peptides from bullfrog skin were evaluated for antitumor activity and improved targeting in melanoma cells. The amino acid sequence of T-La was modified, resulting in the antitumor peptide, T-La (FS). T-La and T-La (FS) were coupled to the RGD small molecule polypeptide to form the chimeric peptides RGD-T-La and RGD-T-La (FS), respectively. The secondary structures for the peptides, evaluated using circular dichroism, were found to be alpha-helical. The structure of T-La was evaluated using bioinformatics. In addition, the antitumor effects of the modified peptide and the targeting of RGD chimeric peptide to the tumor in vivo and in vitro were analyzed. Antitumor activity was measured in vitro using the MTT assay. Tumor cells with high integrin alpha v beta 3 expression were detected using flow cytometry, and tumor cells were screened for sensitivity to RGD-T-La (FS) to establish a tumor model in nude mice. The effects of the peptides on tumor cells were measured using laser confocal microscopy in real-time. The mechanism of the peptide antitumor activity in tumor cells was evaluated with scanning electron microscopy. B16 melanoma cells were the most sensitive to the peptides, for which the cell survival rate was 24.65% for 10 mu g/ml RGD-T-La (FS). RGD-La (FS) had a rapid effect on tumor cells. RGD chimeric polypeptides exhibited site-targeting cytotoxic effects in tumor cells. In the B16 melanoma mouse model, the peptides exhibited antitumor effects against early melanoma development and induced tumor apoptosis, possibly by inhibiting VEGF and promoting caspase-3 expression. Overall, the present study provides a scientific basis for the application of small molecule antimicrobial peptides as targeted antitumor agents and lays the foundation for the clinical application of these peptides as antitumor drugs.
外文关键词：antitumor peptide; RGD-chimera; targeting; antitumor effect; melanoma cells
作者：Liu, MY；Jiang, X；Fu, C；Zhao, RL；Jin, TM；Ma, JF；Qin, SY；Li, LA；Hu, Y；Zhang, X
作者单位：Tianjin Agr Univ
期刊名称：ONCOLOGY LETTERS
期刊影响因子：2.311
出版年份：2021
出版刊次：21
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