[The construction of IGF-II P4 promoter-driven tk expression vector]

Area of Science:

• Molecular Biology
• Gene Therapy
• Oncology

Context:

• Hepatocellular carcinoma (HCC) remains a significant global health challenge.
• Targeted suicide gene therapy offers a promising strategy for cancer treatment.
• The herpes simplex virus-thymidine kinase/ganciclovir (HSV-tk/GCV) system is a well-established suicide gene therapy approach.

Purpose:

• To investigate the selective killing effect of the HSV-tk/GCV suicide gene system.
• To evaluate the efficacy of the system when controlled by the human IGF-II P4 promoter.
• To assess the system's activity specifically in HCC cells (HepG2) versus non-cancerous cells (HeLa) in vitro.

Summary:

• Recombinant shuttle plasmids containing the tkEGFP fusion gene driven by the IGF-II P4 promoter were constructed and transfected into HepG2 and HeLa cells.
• Specific expression of the tkEGFP gene was observed only in HepG2 cells, confirmed by EGFP fluorescence and RT-PCR.
• MTT assays demonstrated significant inhibition of HepG2 cell growth after ganciclovir treatment, while HeLa cell growth remained unaffected, indicating selective toxicity.

Impact:

• Successful construction of a targeted suicide gene vector for HCC.
• Demonstration of selective gene expression and cancer cell killing by the IGF-II P4 promoter-driven HSV-tk/GCV system.
• Provides a strong foundation for developing targeted gene therapy strategies for hepatocellular carcinoma.