Aim: To establish a renal carcinoma cell line which can highly express beta-glucuronidase(betaG), and to observe the biological characteristics of the transfected cells.
Methods: Recombinant eukaryotic expression vector pcDNA3.1-betaG was constructed. It was transfected into renal cancer cells GRC-1 via liposome. The transcription and expression of betaG gene were detected by dot blot and Western blot. The biological characteristics of the betaG gene transfected cells was observed under light microscope, transmission electron microscope and flow cytometry.
Results: Dot blot and Western blot detection confirmed that the betaG gene had been stably integrated into the genomic DNA of the GRC-1 cells and was highly expressed. Transmission electron microscope observation showed that the lysosomes and endoplasmic reticulum were abundant, the number of microvili and process was significantly increased in the transfected cells, but growth condition and cell cycle of GRC-1 cells had no notable difference before and after transfection.
Conclusion: A renal carcinoma cell line that can highly express betaG gene was established, which lays the foundation for further study on gene therapy.