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The Effects of RNA Interference to nm23-M1 Expression on Proliferation of Myeloma Cell Line SP2/0
Qian Ye, Ke Chen, Ming-Yong Luo, Yan-Ling Dong, Ying-Xiong Wang, Jun-Lin He*
Laboratory of Reproductive Biology, Department of Genetics and Eugenics, School of Public Health, Chongqing Medical University, Chongqing 400016, China
Abstract: To investigate the effects of non-metastasis 23-M1 (nm23-M1) gene on proliferation of myeloma cell， we established a myeloma cell line SP2/0 in which nm23-M1 expression was suppressed by RNA interference (RNAi). Three small interfering RNAs (siRNA) targeting nm23-M1 were designed， and three pGenesil-1 plasmids which could express these siRNAs and a negative control plasmid were constructed. Then the recombinant plasmids were transfected into mouse myeloma cell line SP2/0 that clones were selected by G418 in order to establish cell lines SP2/0 with stable expression of pGenesil-1-nm23-M1 siRNA. The expression levels of nm23-M1 mRNA and protein were detected by RT-PCR and Western blot. MTT was used to observe the effects of nm23-M1 gene on proliferation of SP2/0 cells. The results showed three siRNAs could inhibit the expression of nm23-M1 on mRNA and protein levels. Among them， siRNA-2 showed the strongest inhibition effect， with an inhibition rates of 74.4% and 62.1% for the expression of nm23-M1 mRNA and protein respectively. And the proliferation of SP2/0 cells was obviously suppressed after transfection of siRNA-2 (P<0.05). The study constructed pGenesil-1-nm23-M1 siRNA recombination plasmids successfully and screened cell lines SP2/0 in which nm23-M1 expression was inhibited stably. The findings suggested low expression level of nm23-M1 in SP2/0 cells inhibited cell proliferation. It provides a basis for further researching the biological functions of nm23-M1 and related clinical applications.
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