Objective       To construct mCherry reporter system of proteasome maturation protein (POMP) by Crispr/Cas9 and study the impact of POMP on cancer cells.        Methods       POMP-2A-mCherry-T vector was constructed and transferred into human embryonic kidney 293 cells (HEK293 cells). The mCherry-positive cells and mCherry-negative cells were sorted by flow cytometry. Real-time quantitative PCR (RT-qPCR) was used to detect the POMP gene transcription level in the mCherry-positive cells and mCherry-negative cells. Western blotting was adopted to detect the POMP protein expression and JNK1/2 phosphorylation in Huh7 cells. Proteasome activity of mCherry-positive cells and mCherry-negative cells was detected with proteasome activity detection kit.        Results       The protein expression of POMP in the mCherry-positive cells was increased by 1.3 times (P0.01) compared with the mCherry-negative cells

but the POMP gene transcription level had no significant difference between the 2 kinds of cells (P0.05). The level of ubiquitin in the mCherry-negative cells was obviously elevated by 1.2 times than that of mCherry-positive cells(P0.01). The proteasome activity was relatively high in the mCherry-positive cells (P0.01). The JNK1 and of JNK2 phosphorylation levels were increased by 1.5 times and 1.4 times

respectively

in the Huh7 cells with low expression of POMP (P0.01).        Conclusion       The POMP luciferase reporter system is successfully constructed. After the system is applied to Huh7 cells

POMP can change JNK1/2 phosphorylation levels

so it may affect the cell proliferation and apoptosis through JNK signaling pathway.