| Objective:To investigate the effects of non-metastasis 23-M1 ( nm23-M1) gene on proliferation and invasion of myeloma cells, we established a myeloma cell line SP2/0 in which nm23-M1 expression is suppressed by RNA interference ( RNAi) .Method:1 Three small interfering RNAs ( small interfering RNA, siRNA) targeting nm23-M1 gene were designed, and three pGenesil-1 plasmids which express these siRNAs and a negative control plasmid were constructed.2 The recombinant plasmids were transfected into mouse myeloma SP2/0 cells. Then the cell lines SP2/0 which could stably express the designed siRNAs were screened by G418.3 The expression levels of nm23-M1 mRNA and protein were detected by RT-PCR and Western blot in the cell lines SP2/0 which could stably express the designed siRNAs.4 MTT and transwell assays were used to observe the effects of nm23-M1 gene on proliferation and invasion of SP2/0 cells after nm23-M1 gene expression was inhibited.Result:The results showed the three designed siRNAs could inhibit the expression of nm23-M1 on mRNA and protein levels. Among them, siRNA-2 showed the strongest inhibition effect,with inhibition rates of 74.4% and 62.1% for the expression of nm23-M1 mRNA and protein respectively. The proliferation of SP2/0 cells was significantly suppressed after transfection of siRNA-2(P<0.05), and the invasion of SP2/0 cells was not suppressed obviously(P>0.05).Conclusion:1 Three recombinant plasmids expressing three designed siRNAs and a negative control plasmid were successfully constructed.2 The cell lines SP2/0 in which nm23-M1 gene expression could be inhibited stably and efficiently were constructed and screened successfully.3 The results suggested low expression level of nm23-M1 gene in SP2/0 cells inhibited cell proliferation but did not influence cell invasion. It provides a basis for further researching the biological functions of nm23-M1 and related clinical applications. |
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