关键词: 有丝分裂关卡基因, RNA干扰, 细胞增殖, 染色体数目异常, 免疫印迹法

Abstract: Objective To investigate the function of hsMAD2 gene in spontaneous abortion embryos with chromosomal numerical abnormality. Methods The results of quantitative realtime RT-PCR and Western blotting method were used to display the mRNA and protein level of hsMAD2 gene both in spontaneous abortion embryos with chromosomal numerical abnormality (experimental group)and with chromosomal numerical normality (control group). Recombinant shRNA plasmids was constructed targeting hsMAD2 gene to inhibit the expression of endogenous hsMAD2 genes in embryonic cells; interference efficiency was demonstrated by FQ-PCR and Western blotting; cell proliferation was measured by MTT assay；cell cycle was assessed by FCM. Results Western blotting analysis showed that compared with the control group, the protein levels of hsMAD2 in the experimental group decreased significantly.Recombinant shRNA plasmids could significantly and specially inhibit hsMAD2 gene expression in embryonic cells and could inhibit up to 58% of embryonic cells proliferation 48 hours after the transfection. Efficient shRNA can decrease G0/G1 and S phase cells, and increase G2/M phase cells. Conclusion The downexpression of hsMAD2 gene probably plays an important role in the development of spontaneous abortion embryos with chromosomal numerical abnormality. The study provides preliminary data for further establishment of diagnostic or therapeutic criteria on spontaneo

Key words: hsMAD2 gene, RNA interference, Cell proliferation, Numerical chromosomal abnormality, Western blotting