Abstract:

Objective To construct and screen the optimal Caprin⁃1 knockout plasmid and to explore the effect of cell proliferation，cell cycle andthe expression of cyclinD1 and cyclinD2 in hepatocellular carcinoma HepG2 cellsafterCaprin ⁃1⁃KO transfectionmediated byultrasound ⁃microbubbles. Method First of all，Caprin ⁃1 knockout plasmids KO⁃1 and KO⁃2 were constructed and the recombinant plasmids were identified by sequencing. Secondly，after transfecting the target plasmid into HepG2 cells through ultrasound transfection conditions which explored in the previous research，the best quality granules were screened by WB. Finally，qPCR was selected to detect the expression of cyclinD1 and cyclinD2. CCK8 and FCM were used to detect the effect of inhibiting Caprin⁃1 expression on cell proliferation and cycle. Results In this study，the primers of Caprin⁃1 knockout plasmid KO⁃1 and KO⁃2 were designed. The extracted plasmids were constructed successfully after electrophoresis and sequenc⁃ ing. The results showed that the expression of Caprin⁃1 in NC group and KO⁃1 group were higher compared to the KO⁃2 group（P<0.05）.After inhibiting the expression of Caprin⁃1，the proliferation of HepG2 cells was significantly inhibited，and the cell cycle was blocked；the expression of cyclinD1 and cyclinD2 was significantly down⁃regulat⁃ ed（P<0.05）. Conclusion Caprin⁃1⁃KO were transfected into HepG2 cells by UTMD. The expression of Caprin⁃1⁃ KO can inhibit the cell proliferation、block the cycle and decrease the expression of cyclinD1 and cyclinD2. This study is expected to provide ideas for further research on the role of Caprin⁃1 in the pathogenesis of liver cancer. 

Key words:

Caprin?1, ultrasound microbubble,