HSV-TK/IL-12联合基因疗法通过Cx43上调及免疫激活引发的旁观者效应，抑制非小细胞肺癌的生长。

Abstract

GCV reduces the survival rate of NSCLC cells. • The combination gene therapy of HSV-TK and IL-12 induces the apoptosis of NSCLC cells. • The combined HSV-TK/IL-12 gene therapy enhances lymphocyte proliferation. • HSV-TK/IL-12 combination therapy exerts a bystander effect on NSCLC cells. • This combined strategy effectively suppresses NSCLC tumor growth and experimental lung colonization in a tail-vein injection model, demonstrating its potential as a novel gene therapy to overcome current clinical treatment challenges. Background Lung cancer remains the leading cause of cancer-related mortality worldwide, with non-small cell lung cancer (NSCLC) representing a major subtype characterized by high incidence and mortality. Despite advances in targeted therapy and immunotherapy, treatment resistance and the immunosuppressive tumor microenvironment remain major clinical challenges. The combination of herpes simplex virus thymidine kinase and interleukin-12 (HSV-TK/IL-12) has demonstrated anti-tumor potential, but its role and mechanism of action in NSCLC remain poorly defined. This study aims to systematically evaluate the anti-tumor efficacy of HSV-TK/IL-12 combination gene therapy in NSCLC and to investigate the potential involvement of Connexin 43 (Cx43) in the bystander effect (BSE) alongside immune activation. Methods Overexpression vectors for HSV-TK and IL-12 were constructed to study their functions in NSCLC. Protein expression was measured by western blot. 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl tetrazolium bromide (MTT) assay and flow cytometry were employed to assess cell survival and apoptosis, respectively. In vitro , human peripheral blood lymphocytes were used to evaluate immune responses relevant to clinical immunotherapy. In vivo , xenograft mouse models were used to evaluate effects on tumor growth and experimental lung colonization. Results Overexpression of HSV-TK or IL-12 significantly increased their respective protein levels in A549-TK and H12