Effects of folate deficiency on proliferation, apoptosis and expression of spindle assembly checkpoint-related proteins in L02 cells

ObjectiveTo investigate the effect of folate deficiency on the proliferation, apoptosis, expression of spindle assembly checkpoint-related proteins, and its mechanism in L02 cells.

MethodsL02 cells were cultured with different concentrations of folate(2 000, 200, 20, 0 nmol/L) for 2 weeks.Then the morphology of L02 cells was observed by inverted microscope, the cell cycle and apoptosis were detected by flow cytometry, cell proliferation was measured by CCK8, the expressions of Mad2, Bub1, BubR1 mRNA were detected by fluorescence quantitative PCR, and the expressions of Mad2, Bub1, BubR1 protein were analyzed by Western blotting.

ResultsCompared with the normal control group, folate deficiency made L02 cells to become larger and irregular in shape, reduced the cell viability(P < 0.01), induced S-phase block(P < 0.05), promoted apoptosis(P < 0.05), and upregulated the mRNA and protein expressions of Mad2, Bub1, BubR1 in L02 cells(P < 0.01).

ConclusionsIn the culture L02 cells, it induces folate deficiency at 200 nmol/L folate, which causes impaired spindle assembly checkpoint function, abnormal cell proliferation, and even apoptosis in severe deficiency.