ObjectiveTo explore the relationship between DJ-1, DJ-1M26I, DJ-1L166P with the cell proliferation and apoptosis of NIH 3T3 cells at cellular level, and provide a basis for the construction of a transgenic animal model of Parkinson’s disease and further study on the pathogenesis of this disease. MethodsRecombinant plasmids pcDNA3.1/myc-His-DJ-1, pcDNA3.1/myc-His-DJ-1L166P and pcDNA3.1/myc-His-DJ-1M26I were transfected into NIH 3T3 cells, respectively, using lipofectamine. The cells were screened with G418 at a dose of 500 μg/mL. Stable clones were identified on the DNA, RNA and protein levels. MTT assay and annexin V-FITC kit were used to detect the viability and apoptosis of those stable cell clones. ResultsAfter the G418-screening of of NIH 3T3 cells transfected with recombinant plasmids pcDNA3.1/myc-His-DJ-1, pcDNA3.1/myc-His-DJ-1L166P or pcDNA3.1/myc-His-DJ-1M26I, one, four and three positive clones were obtained, respectively, by PCR detection. RT-PCR and Western blot detected the expression of DJ-1-His in the positive clones. NIH 3T3 cells transfected with DJ-1L166P and DJ-1M26I had a higher expression of caspase-3 mRNA than normal NIH 3T3 cells, while NIH3T3 cells transfected with DJ-1 had a lower expression. MTT assay showed that NIH 3T3 positive cells transfected with DJ-1L166P and DJ-1M26I had a lower proliferation rate than that of normal NIH3T3 cells (P<0.05), while the NIH 3T3 positive cells carrying DJ-1 gene did not show significant difference compared with the normal NIH 3T3 cells. Apoptosis test indicated that the apoptosis rates of DJ-1L166P and DJ-1M26I transfected cells were higher than that of normal NIH 3T3 cells, however the apoptosis rate of the DJ-1-transfected cells was significantly lower than that of normal NIH 3T3 cells (P<0.05). ConclusionsDJ-1L166P and DJ-1M26I mutations reduce the proliferation of NIH 3T3 cells. DJ-1L166P and DJ-1M26I mutations also enhance apoptosis in NIH 3T3 cells. Their effects on NIH 3T3 cell proliferation and apoptosis are similar.