人Makorin环指蛋白1基因沉默对HEK293细胞的影响

Effect of Sliencing Human Makorin Ring Finger Protein 1 Gene on HEK293 Cells

目的探讨特异性抑制人Makorin环指蛋白1(MKRN1)基因的表达对HEK293细胞的影响。方法用脂质体将前期筛选出的含最有效干扰序列的人MKRN1基因shRNA真核表达质粒,转染至HEK293细胞中,观察其对人端粒酶逆转录酶(hTERT)基因mRNA转录水平、蛋白表达水平及细胞增殖的影响。结果人MKRN1基因shRNA真核表达质粒转染HEK293细胞后,在mRNA和蛋白水平均能明显上调细胞hTERT基因的表达,且细胞明显增殖。结论抑制HEK293细胞中人MKRN1基因的表达,可导致hTERT基因mRNA转录水平和蛋白表达水平上调,并促进细胞增殖。

Objective To investigate the effect of specifically inhibiting the expression of human Makorin ring finger protein 1 （MKRN1）gene on HEK293 cells. Methods Previously constructed eukaryotic expression vector for short hairpin RNA（shRNA）of human MKRN1, containing the most efficient interfering sequence, was transfected to HEK293 cells, and the effects on transcription of human telomerase reverse transcriptase （hTERT） mRNA, expression of hTERT protein and proliferation of HEK293 cells were ob-served. Results Both the hTERT mRNA transcription and protein expression levels in HEK293 cells after transfection with the con-structed recombinant plasmid, as well as the proliferation of HEK293 cells, increased significantly as compared with those before tran-fection. Conclusion The inhibition of expression of human MKRN1 gene in HEK293 cells up-regulated the hTERT mRNA tran-scription and protein expression and promoted the proliferation of HEK293 cells.