Construction of lentiviral vector with overexpression of ERp29 and its effect on biological behavior of gastric cancer cells / 吉林大学学报(医学版)

ABSTRACT

Objective: To construct a lentiviral vector with overexpression of endoplasmic reticulum protein 29 ERp29), and to explore the effects of ERp29 overexpression on the proliferation, migration and invasion of gastric cancer MGC803 and SGC7901 cells. Methods: The lentiviral plasmid with ERp29 pCDH-ERp29) tagged with red fluorescentce protein and control plasmid pCDH-Vector) were constructed, and then the MGC803 and SGC9901 cells were infected with these lentivital vectors. Under fluorescence microscope, the infection of these cells were observed. CCK-8 assay and clone formation assay were performed to test the proliferation abilities of MGC803 and SGC7901 cells infected with ERp29. Transwell chamber assay was used to test the abilities of migration and invasion of MGC803 and SGC7901 cells infected with ERp29. Real-time PCR and Western blotting methods were used to detect the expression levels of ERp29 mRNA and protein in MGC803 and SGC7901 cells which were stably infected by ERp29, respectively. Results: The enzyme digestion identification result showed that the pCDH-ER29 overexpression vector was successfully constructed. The fluorescence microscope result showed the successful infection in the gastric cancer cells. The CCK-8 assay and clone formation assay results showed that compared with pCDH-Vector group, the proliferation ability of the cells in pCDH-ERp29 group had no marked changes (P > 0.05). Transwell chamber assay revealed that compared with pCDH-Vector group, the number of migration and invasion cells in the MGC803 and SGC7901 cells in pCDH-ERp29 group were significantly decreased (P < 0 . 05). The expression levels of ERp29 mRNA and protein in the cells in pCDH-ERp29 group were significantly increased compared with pCDH-Vector group (P < 0 . 05). Conclusion: The lentiviral vector with overexpression of ERp29 is constructed successfully, and the overexpression of ERp29 in MGC803 and SGC7901 cells can significantly inhibit their abilities of migration and invasion.