| In recent years,the incidence of cancer is higher than before.Finding the causes of cancer and effective treatment methods to prolong the patients' life and cure cancer patients completely has become the focus of current research.Currently,except traditional surgical treatments,radiotherapy and chemotherapy,immunotheapy and gene therapy are considered as the most promising methods to overcome cancer.Among them,CAR-T therapy is the mainstream in immunotherapy research,and targeted gene-virus targeted therapy is the focus of gene therapy research,several combination therapies are also under development and research,which requires us to have a clearer understanding of the effects and side effects of each method.Our group has been studied the effect of lectin genes in cancer gene therapy,using oncolytic vaccinia virus and adenovirus as vectors for a long time,and has found several effective lectin genes can promote apoptosis of tumor cell.We found Haliotis discus discus sialic acid binding lectin(HddSBL)has a unique role in the study of glioma,so we did a further study to understand the effect of HddSBL in glioma cells.In this study,we first constructed a recombinant oncolytic vaccinia virus named oncoVV-HddSBL,using PBS and empty oncolytic vaccinia virus named oncoVV as control,and study the effect of HddSBL in the in vivo treatment of C6 cell Xenografts tumor in nude mice.The results showed that oncoVV caused the death of mice,and HddSBL prolonged more than nine and a half days of survival time of mice;Using semi-quantitative PCR detected the expression of related inflammatory factor in C6 cells after viral treatment,and we found oncoVV increased the level of IL-1?,IL-6and TNF-?.Combining the phenomenon of weight loss before death of mice,we considered that oncoVV caused mice tumor cachexia,while HddSBL can reduce the expression of IL-6 and TNF-?,alleviated cachexia;Using dual luciferase reporter genes detected NF-?B and AP-1 inflammation-related transcription factors revealed that HddSBL did not inhibit NF-?B and AP-1 activity to reduce the expression ofrelated inflammatory factors.We detected IL-2 levels of rat in mice serum by ELISA,and found that oncoVV caused a high expression of IL-2 in C6 cells,while HddSBL could reduce the expression of IL-2,which may be the way of HddSBL reducing the expression of inflammation to alleviat cachexia and prolong the life of mice;To further study the role of HddSBL in C6 cells,we performed transcriptome sequencing and analysis.We have a keen interest that HddSBL affected multiple histone components and reduced multiple antiviral genes.Detection of related proteins by western blot revealed that HddSBL significantly decreased Histone H3 and Histone H4 protein levels,while Histone H3 8th arginine asymmetric dimethylation and Histone H4 3rd arginine asymmetric dimethylation increased significantly,which indicated HddSBL altered the state of histones,affecting chromatin nucleosome assembly,and participateed in the regulation of chromatin levels.This result was consistent with the results of transcriptome analysis.We used semi-quantitative PCR for related antiviral genes.The results showed that oncoVV increased the level of MX2,DDX58,IFIT3 and IFIT2 antiviral genes,while HddSBL reduced the level of MX2,DDX58,IFIT3 and IFIT2,which was consistent with the results of transcriptome sequencing.Moreover,we detected the replication of oncoVV and oncoVV-HddSBL in C6 cells and found that HddSBL promoted the replication of oncolytic vaccinia virus significantly in C6 cells,which may caused by downregulation of antiviral protein.In this study,we used oncolytic vaccinia virus as a carrier,providing some theoretical and research basis for HddSBL reduce the toxicity of oncolytic vaccinia virus in animal models of glioma. |
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