CN-122011153-A - Short peptide, slow virus and cell expressing short peptide and application thereof

Abstract

The invention provides a short peptide, and a lentiviral expression vector and a recombinant cell for expressing the short peptide. And further provides its use in the treatment of tumors. After the short peptide is delivered to HT1080 cells through a lentiviral system, the intracellular SLC7A11 protein level is down regulated, the intracellular Fe 2+ and lipid peroxidation level are obviously increased, the short peptide has obvious inhibition effect on cell proliferation, and the short peptide has tumor treatment potential.

Inventors

• WU GUIXIAN
• ZHOU YUANZE
• LUO CHENGXIN
• XU SHUANGNIAN

Assignees

• 中国人民解放军陆军军医大学第一附属医院

Dates

Publication Date

20260512

Application Date

20260128

Claims (9)

1. 1. A short peptide has an amino acid sequence of SEQ ID NO. 1.
2. 2. The gene encoding the short peptide according to claim 1, wherein the nucleotide sequence is SEQ ID NO. 2.
3. 3. A recombinant lentiviral expression vector comprising the coding gene of claim 2.
4. 4. A recombinant lentiviral expression vector according to claim 3, comprising a CMV promoter, preferably pLVX lentiviral vectors.
5. 5. A recombinant cell comprising a gene encoding the recombinant lentiviral expression vector of claim 3 or 4; preferably, the recombinant cell is prepared by a method comprising the steps of: The recombinant lentiviral expression vector of claim 3 or 4, together with packaging plasmid psPAX and pMD2G, is co-transfected into HEK293T cells for 48-72 hours, the culture medium containing the viral particles is collected, the culture medium containing the viral particles is co-cultured with target cells for 48 hours, and cell clones with stable expression are selected by puromycin.
6. 6. A pharmaceutical composition comprising the short peptide of claim 1, the recombinant lentiviral expression vector of claim 3 or 4, or the recombinant cell of claim 5.
7. 7. Use of the short peptide of claim 1, the recombinant lentiviral expression vector of claim 3 or 4, or the recombinant cell of claim 5 in the manufacture of a medicament for treating a tumor.
8. 8. The use of claim 7, wherein the tumor is selected from one or more of Fibrosarcoma (FS), sarcoma (SARC), breast cancer (BRCA), hepatocellular carcinoma (LIHC) or pancreatic cancer (PAAD).
9. 9. The use according to claim 8, wherein the tumour is fibrosarcoma.

Description

Short peptide, slow virus and cell expressing short peptide and application thereof Technical Field The invention relates to the technical field of biological medicines, in particular to application of an SLC7A11 derivative peptide FP-01 as an iron death inducer in tumor treatment. Background Iron death (Ferroptosis) is an iron ion dependent apoptosis characterized by lipid peroxidation and glutathione depletion. Iron death is morphologically characterized by mitochondrial atrophy, increased membrane density and decreased mitochondrial cristae, and is mechanistically regulated by various pathways such as the cystine-glutamate antiport protein System (System XC-), the mevalonate pathway and the mitochondrial DHODH. Iron death is a novel cell death mode, and biological effects after activation or inhibition are widely focused and studied. The iron death inhibitor can protect normal tissues by inhibiting iron death, is a potential or auxiliary treatment means for aging-related injury diseases such as acute kidney injury, acute liver injury, radiation injury, neurodegeneration and the like, and can directly induce partial tumor cells to generate iron death or combine with clinical tumor medicaments to achieve the effect of cooperatively treating tumors. However, toxicity problems and ambiguous mechanisms of the related formulations, whether iron death inhibitors or iron death activators, are important obstacles to their clinical transformation. The cystine-glutamate antiport protein System (System XC-) is a dimer inlaid in cell membranes, consists of SLC3A2 and SLC7A11, and is one of the main mechanisms for regulating cell iron death. SLC7A11 is the main subunit of System XC-function, and is highly expressed in various cancer cells, and the survival time of a patient with high expression of SLC7A11 is obviously lower than that of a patient with low expression, which indicates that the SLC7A11 with high expression is a bad prognosis factor of tumor. Mechanically, overexpression of SLC7a11 causes increased biosynthesis of reduced Glutathione (GSH), which in turn inhibits the occurrence of lipid peroxidation, thereby inhibiting iron death of the cells and promoting tumor growth. Thus, targeted modulation of the SLC7a11 or System XC-System is a potential approach to tumor treatment. SLC7A11 is a multiple transmembrane protein consisting of the N-terminal (1-43), C-terminal (471-501) and core transmembrane region (44-470) located in the cytoplasmic region. Disclosure of Invention The invention aims to provide a short peptide drug (FP-01) with iron death activity and application thereof in tumor treatment. The short peptide medicine FP-01 is used as an iron-death promoting preparation, can promote iron death of tumor cells by downregulating the protein level of core subunit SLC7A11 in a cystine-glutamic acid antiport protein System (System XC-) path, and further relieves tumor-bearing pressure of mice tumors, and has the potential of tumor treatment. The invention firstly provides a short peptide, the amino acid sequence of which is SEQ ID NO. 1. The invention further provides a coding gene of the short peptide, and the nucleotide sequence of the coding gene is SEQ ID NO. 2. The invention provides a recombinant lentiviral expression vector, which comprises the coding gene. In one embodiment according to the invention it comprises a CMV promoter, preferably pLVX lentiviral vectors. The invention also provides a recombinant cell which comprises the coding gene of the recombinant lentiviral expression vector; in one embodiment according to the invention, the recombinant cell is prepared by a method comprising the steps of: the recombinant lentiviral expression vector is co-transfected with a packaging plasmid psPAX and pMD2G into HEK293T cells for 48-72 hours, a culture medium containing virus particles is collected, the culture medium containing virus particles is co-cultured with target cells for 48 hours, and cell clones with stable expression are screened out through puromycin. In yet another aspect, the invention provides a pharmaceutical composition comprising the above-described short peptide, recombinant lentiviral expression vector, or recombinant cell. In another aspect, the invention provides the use of the above-described short peptide, recombinant lentiviral expression vector, or recombinant cell in the preparation of a medicament for treating a tumor. In one embodiment according to the invention, the tumor is selected from one or more of Fibrosarcoma (FS), sarcoma (SARC), breast cancer (BRCA), hepatocellular carcinoma (LIHC) or pancreatic cancer (PAAD). In one embodiment according to the invention, the tumor is fibrosarcoma. The technical scheme of the invention has the following beneficial effects: The short peptide FP-01 provided by the invention is an active peptide capable of inducing cells to generate iron death, can promote iron death of tumor cells by downregulating the protein level of core subunit