Preparation of a novel recombinant vaccinia virus carrying CXCL9 and IL-7 genes and its anti-tumor effect in vitro

Corresponding author: Zhanglin, stepinghuns2@163.com

DOI: 10.12201/bmr.202607.00010

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  Abstract: Objective: To prepare a novel recombinant vaccinia virus carrying CXCL9 and IL-7 double genes by molecular biological method, and to preliminarily verify the effect of the recombinant vaccinia virus in infecting and killing gastric cancer cells by in vitro cytological experiments. Methods: The recombinant vaccinia virus eukaryotic expression plasmid pSC65-CXCL9-IL7 carrying CXCL9 and IL-7 double genes was constructed step by step, and then the vaccinia virus strain was recombined in eukaryotic cells with this vector. After repeated screening and purification with limited dilution method, purified recombinant oncolytic poxvirus VV- CXCL9-IL7 was obtained, Then, the recombinant oncolytic poxvirus with high titer was obtained by infecting HeLa cells to expand the culture and using cesium chloride gradient centrifugation to concentrate and purify it. The recombinant oncolytic poxvirus was used to infect human gastric cancer cell line SGC7901 in vitro. The infectivity of recombinant virus was detected by TCID50 method, and the concentration of CXCL9 and IL-7 in cell culture supernatant was detected by ELISA. The cytotoxicity of VV-CXCL9-IL7 to SGC7901 cells was detected by MTT method. The effect of recombinant oncolytic poxvirus infection on cell growth was detected by cell growth curve method, and the changes of cell cycle and apoptosis of infected SGC7901 cells were detected by flow cytometry. Results: The recombinant oncolytic poxvirus VV-CXCL9-IL7 was successfully constructed, and the production and release of chemokines IL-7 and CXCL9 were preliminarily verified by the detection of the concentration of supernatant. The results of cell infection intervention showed that the killing effect of tumor cells in the VV-CXCL9-IL7 recombinant oncolytic poxvirus intervention group was significantly stronger than that in each control group, and the tumor growth was significantly slower than that in other control groups. The G0/G1 phase ratio of cells in the oncolytic poxvirus intervention group was significantly higher than that in each control group, and its apoptosis rate was significantly increased. Conclusion: In vitro cytological intervention experiment, VV-CXCL9-IL7 recombinant infection has a strong anti-tumor effect on gastric cancer cells, and has potential clinical application value for gastric cancer, which can be infected by local direct drug administration.

  Key words: gastric cancer; Oncolytic poxvirus; CXCL9; IL-7; Bioimmunotherapy

  Submit time: 26 August 2026

  Copyright: The copyright holder for this preprint is the author/funder, who has granted biomedRxiv a license to display the preprint in perpetuity.
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