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目的观察NIRF蛋白对妊娠早期绒毛滋养细胞增殖、凋亡的影响及其作用机制。方法取正常妊娠妇女人工流产绒毛组织(8~10周),按常规分离方法获得滋养细胞,进行原代培养。通过上/下调NIRF在滋养细胞中的表达后,采用qRT-PCR法检测p53基因mRNA的转录水平;Western blot法检测p53蛋白的表达水平;MTT法检测滋养细胞的增殖能力;Annexin V/PI双染色法检测细胞凋亡情况。结果与未转染组比较,NIRF上/下调后,p53基因m RNA转录水平差异无统计学意义(P>0.05);NIRF上调,p53蛋白表达水平明显下降(P<0.05),早期绒毛滋养细胞增殖能力显著增强(P<0.05),凋亡率差异无统计学意义(P>0.05);NIRF下调,p53蛋白的表达水平明显升高(P<0.05),早期绒毛滋养细胞增殖能力明显减弱(P<0.05),凋亡率明显上升(P<0.05)。结论 NIRF在蛋白水平上抑制了p53的表达,促进了滋养细胞的增殖。
Objective To observe the effect of NIRF protein on the proliferation and apoptosis of villus trophoblast during early pregnancy and its mechanism. Methods Normal pregnant women with artificial abortion villi (8 to 10 weeks), according to the conventional separation methods to obtain trophoblast cells for primary culture. The transcription level of p53 mRNA was detected by qRT-PCR after up / down-regulating the expression of NIRF in trophoblast cells. Western blot was used to detect the expression of p53 protein. The proliferation of trophoblast cells was detected by MTT assay. Annexin V / Staining assay apoptosis. Results Compared with the untransfected group, there was no significant difference in the m RNA transcription level of p53 gene after up / down regulation of NIRF (P> 0.05); NIRF upregulation, p53 protein expression decreased significantly (P <0.05), early villous trophoblast (P <0.05). There was no significant difference in apoptotic rate between the two groups (P> 0.05). The expression of p53 protein was significantly increased (P <0.05) and the proliferation of early trophoblast cells was significantly decreased P <0.05), the apoptosis rate increased significantly (P <0.05). Conclusion NIRF inhibits the expression of p53 at the protein level and promotes the proliferation of trophoblasts.