Construction of an oral squamous cell carcinoma cell line for stable PA28γ overexpression

• Published in: Hua xi kou qiang yi xue za zhi = Huaxi kouqiang yixue zazhi = West China journal of stomatology Vol. 38; no. 1; p. 6
• Main Authors: Xin, Chuan, Wang, Jiong-Ke, Li, Jing, Zeng, Xin
• Format: Journal Article
• Language: Chinese
• Published: China 01.02.2020
• Subjects: Autoantigens; Carcinoma, Squamous Cell; Cell Line, Tumor; Genetic Vectors; Humans; Lentivirus; Mouth Neoplasms; Proteasome Endopeptidase Complex; Transfection; stable overexpression cell line; PA28γ; oral squamous cell carcinoma; overexpression vector; lentivirus
• ISSN: 2618-0456

To construct a PA28γ overexpression cell line and determine its effects after infecting an oral squa-mous cell carcinoma (OSCC) cell line. The PA28γ 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 PA28γ overexpression. Finally, the level of PA28γ expression in the OSCC cell line was detected through Western blot. The successful construction of PA28γ recombinant lentiviral vectors was confirmed by DNA sequencing. The results of immunofluorescence showed that the PA28γ overexpression lentivirus successfully infected the OSCC cells and showed cherry red fluorescence. The results of Western blot demonstrated that the constructed cells with stable PA28γ overexpression significantly increased the expression of PA28γ. The PA28γ overexpression lentiviral vector ca