Construction of an oral squamous cell carcinoma cell line for stable PA28gamma overexpression

OBJECTIVE: To construct a PA28gamma overexpression cell line and determine its effects after infecting an oral squa-mous cell carcinoma (OSCC) cell line. METHODS: The PA28gamma gene was cloned into the pLOV.CMV.cherry.2A.EF1a.PuroR lentiviral vector by polymerase chain reaction (PCR), and PCR and DNA sequencing alignment analysis were used for identification. Then, 293T cells were used to package viral diseases. Infected OSCC cells were used to construct a cell line with stable PA28gamma overexpression. Finally, the level of PA28gamma expression in the OSCC cell line was detected through Western blot. RESULTS: The successful construction of PA28gamma recombinant lentiviral vectors was confirmed by DNA sequencing. The results of immunofluorescence showed that the PA28gamma overexpression lentivirus successfully infected the OSCC cells and showed cherry red fluorescence. The results of Western blot demonstrated that the constructed cells with stable PA28gamma overexpression significantly increased the expression of PA28gamma. CONCLUSIONS: The PA28gamma overexpression lentiviral vector can significantly increase its protein expression in OSCC cells. We provide a stable OSCC cell line for further study on the effect of PA28gamma in OSCC.