Background: Antimicrobial and antifungal activities of Thrombocidin-1 (TC-1) is shown previously, however,the anti-cancerous feature of this peptide is still uncovered.

Objective: The objective is to evaluate anti-cancerous feature of recombinant TC-1.

Methods: In this study, based on the significant similarity of rTC-1 and IL-8 in case of coding sequence, tertiary structure, and also docking and molecular dynamic simulation (MD) results with CXCR1, a receptor which has positive correlation with different cancers, a likely pathway for anticancerous effect of rTC-1 was proposed. In addition, the coding sequence of TC-1+6xhistidine (rTC-1) was inserted into the pET22b(+) vector and cloned and expressed by E. coli BL21 and finally purified through nickel affinity column. Afterward, the retrieved rTC-1 was used in MTT assay against mouse colon adenocarcinoma, hepatocellular carcinoma, chondrosarcoma, mouse melanoma, and breast adenocarcinoma cell lines to investigate its probable anticancer application.

Results: Docking and MD simulation results showed that rTC-1 and IL-8 share almost the same residues in the interaction with CXCR1 receptor. Besides, the stability of the rTC-1_CXCR11-38 complex was shown during 100ns MD simulation. In addition, the successful expression and purification of rTC-1 depict an 8kD peptide. The IC50 results of MTT assay revealed that rTC-1 has cytotoxic effect on C26-A and SW1353 cancerous cell lines.

Conclusion: Therefore, apart from probable anti-cancerous effect of rTC-1 on C26-A and SW1353 cell lines, this peptide may be able to mimic the anti-cancerous pathway of IL-8.

背景：先前已显示了血栓蛋白1（TC-1）的抗菌和抗真菌活性，但该肽的抗癌特性仍未发现。

目的：目的是评估重组TC-1的抗癌特性。

方法：本研究基于rTC-1和IL-8在编码序列，三级结构以及与CXCR1的对接和分子动力学模拟（MD）结果方面的显着相似性，该受体与不同的受体具有正相关对于癌症，有人提出了rTC-1抗癌作用的可能途径。此外，将TC-1 + 6xhistidine（rTC-1）的编码序列插入pET22b（+）载体中，并通过大肠杆菌BL21克隆和表达，最后通过镍亲和柱纯化。之后，将回收的rTC-1用于针对小鼠结肠腺癌，肝细胞癌，软骨肉瘤，小鼠黑素瘤和乳腺腺癌细胞系的MTT分析，以研究其可能的抗癌应用。

结果：对接和MD模拟结果表明，rTC-1和IL-8在与CXCR1受体的相互作用中共有几乎相同的残基。此外，在100ns MD模拟过程中显示了rTC-1_CXCR11-38复合体的稳定性。另外，rTC-1的成功表达和纯化显示了一个8kD肽。MTT分析的IC50结果表明，rTC-1对C26-A和SW1353癌细胞系具有细胞毒性作用。

结论：因此，除了rTC-1对C26-A和SW1353细胞系可能具有抗癌作用外，该肽可能还可以模拟IL-8的抗癌途径。