Construction and functional analysis of luciferase reporter plasmid containing CXCL12 Gene 3’UTR

Inhalt:
The morbidity and mortality of gastrointestinal cancer increase yearly. Previous studies have shown that CXCL12 promotes tumour growth and malignancy, enhances tumour angiogenesis, participates in tumour metastasis. MiRNAs are short, non-coding RNAs that inhibiting protein translation or inducing mRNA degradation by targeting the 3'UTR of mRNA. CXCL12 can become an important target for the development of novel anticancer therapies through the miRNAs pathway. However, some of the regulatory mechanisms of the CXCL12 gene are still unclear. In this study, human CXCL12 3’UTR luciferase reporter plasmid was successfully constructed. Then co-transfected with CXCL12 3’UTR luciferase reporter plasmid and miR-5100 mimics or negative control into HEK293T cells, analysis the luciferase activity 48 hours after transfection, the transfection efficiency was confirmed by transfection of green fluorescent protein GFP. The result showed that transfection of miR-5100 mimics can decrease the luciferase activity of CXCL12 3’UTR luciferase reporter plasmid compared with the negative control group. Our study reveals a regulatory pathway for the CXCL12 gene involved in microRNAs, providing a theoretical basis for the clinical treatment of gastric cancer and the development of new drugs.