[Effect of GRK6 on Proliferation of Multiple Myeloma MM1R Cells and Its Mechanism].

OBJECTIVE

To explore the regulatory effect of GRK6 on the proliferation of multiple myeloma cells and its mechanism.

METHODS

A lentivirus vector shRNA interfering in human GRK6 gene expression was constructed and trans-fected into multiple myeloma cells to obtain the cell line MM1R with stable down-regulation of GRK6 gene expression. The real-time quantitative PCR and Western blot were used to confirm the effectiveness of the GRK6 gene expression down-regulation mediated by lentivirus vector. The MM1R cells with most obvious down-regulation were selected to detect the effect of GRK6 gene on cell proliferation.

RESULTS

The lentivirus vector GRK6-shRNA interfering in human GRK6 gene was constructed succesufully and transfected into multiple myeloma cells, thereby the MM1R cell line with stable down-regulation of GRK6 gene was obtained. The CCK-8 assay showed that the proliferative viability of MM1R cells in experimental group was significantly lower than that in control group (P<0.05); the flow cytometry showed that cells in experimental group were arrested in G/G phase(P<0.05); the Western blot detection showed that the Cyclin D1 and CDK4 levels in experiment group obviously decreased as compared with control group.

CONCLUSION

A lentivirus vector which can specifically interfere in GRK6 gene expression is constructed successfully, The MM1R cell line with stable down-regulation of GRK6 expression is obtained by transfection and screening. The down-regulation of GRK6 expression can arrest MM1R cells in G/G phase, moreover inhibits the proliferation of MM1R cells by inhibition of Cyclin D1 and CDK4 levels.