Cancer remains one of the leading causes of mortality across the globe, despite decades of intensive research and therapeutic advancements [1, 2]. While conventional approaches such as chemotherapy, radiotherapy, and targeted therapies have improved survival in certain malignancies, they are frequently associated with serious side effects, tumor resistance, and limited efficacy in advanced stages [3,4,5]. Gastric cancer, in particular, is a substantial therapeutic challenge owing to its aggressive course, late diagnosis, and poor five-year survival rate [6,7,8]. Gastric cancer ranks as the fifth most common malignancy and the third highest contributor to cancer-related deaths globally, highlighting the urgent need for more precise and effective treatment approaches [9,10,11]. Among its subtypes, poorly differentiated gastric carcinoma—represented by cell lines such as HGC-27—is particularly difficult to treat and often resistant to standard modalities.

In recent years, MSCs have gained attention as potential therapeutic carriers for cancer therapy due to their natural tumor-homing ability, low immunogenicity, and suitability for genetic modification [12]. Mesenchymal stem cells (MSCs), sourced from tissues such as adipose tissue, bone marrow, and umbilical cord, possess the ability to migrate toward tumor microenvironments (TMEs) and deliver therapeutic agents [13]. Genetic modification of MSCs permits the expression of therapeutic transgenes such as cytokines, pro-apoptotic genes, and immunomodulatory agents, enabling localized anti-tumor effects with reduced systemic toxicity compared with conventional cytokine therapies [14,15,16]. Preclinical studies have demonstrated that engineered MSCs can effectively migrate to tumor sites and deliver targeted therapeutic payloads, resulting in tumor growth inhibition in various solid tumor models.

HUC-MSCs are especially attractive owing to their non-invasive source, higher proliferation capacity, and robust immunomodulatory functions. Several preclinical studies have shown the utility of MSCs in delivering pro-apoptotic genes, cytokines, and oncolytic viruses directly to tumor sites [17]. However, clinical translation remains limited by its insufficient tumor targeting, risk of tumor promotion under certain conditions, and inadequate stimulation of anti-tumor immunity [18]. These limitations underscore the need for genetic enhancement of MSCs to fully realize their potential in cancer therapy.

One key avenue of improvement lies in enhancing the tumor-homing capabilities of MSCs through the overexpression of specific chemokine receptors. Among these, C-X-C chemokine receptor type 4 (CXCR4) has been widely studied for its role in the SDF-1/CXCR4 axis, which is instrumental in regulating cell migration and retention within the tumor microenvironment [17]. Many solid tumors, including gastric cancer, exhibit elevated levels of stromal cell-derived factor-1 (SDF-1), which attracts CXCR4-expressing cells to the tumor site. Engineering HUC-MSCs to overexpress CXCR4 has shown promise in improving their homing efficiency and persistence at the tumor location, thereby increasing the therapeutic payload delivered to malignant tissues.

While CXCR4 enhances localization, TNFSF14—also known as LIGHT (homologous to lymphotoxins, exhibits inducible expression)—acts as a potent immunomodulatory molecule. By competing with HSV glycoprotein D for Herpesvirus Entry Mediator (HVEM), a receptor expressed on T lymphocytes, LIGHT can effectively trigger anti-tumor immune responses [19]. LIGHT, a member of the TNF superfamily, binds to two key receptors—HVEM and Lymphotoxin Beta Receptor (LTβR)—thereby stimulating T cell activation, enhancing dendritic cell function, and promoting the destruction of tumor vasculature. It has also been shown to reshape the immunosuppressive tumor microenvironment and increase infiltration of cytotoxic T lymphocytes [20]. When expressed by MSCs, LIGHT has the potential to convert these otherwise immunosuppressive cells into agents of immune activation within the tumor niche. Thus, LIGHT represents a desirable therapeutic gene for inclusion in engineered MSC-based cancer therapies [20].

Based on these insights, we hypothesize that dual genetic modification of HUC-MSCs with CXCR4 and TNFSF14 can synergistically enhance tumor targeting and immunogenicity, thereby providing a potent therapeutic platform for gastric cancer. In this study, we established a lentiviral vector-based transduction system to co-express CXCR4 and TNFSF14 in HUC-MSCs and evaluated their anti-tumor effects against the HGC-27 gastric cancer cell line. This novel approach integrates the migratory advantage conferred by CXCR4 with the immune-stimulatory potential of LIGHT, potentially transforming MSCs into an efficient and targeted system for tumor immunotherapy. To our knowledge, this is among the first studies to investigate the combined therapeutic potential of CXCR4 and TNFSF14 in genetically engineered MSCs, specifically against gastric cancer.