1.Over-expression of circRNA ubiquitin ligase E3 (circHERC4/hsa_circ_0007113) inhibits cell senescence
Hualing Li ; Di Yang ; Junjie Wei ; Zhangyue Chen ; Juping Chen
Acta Universitatis Medicinalis Anhui 2022;57(7):1100-1105
Objective:
To investigate the relationship between the ubiquitin ligase E3 cirRNA(circHERC4/hsa_circ_0007113) and the senescence in Human embryonic lung fibroblasts(IMR-90)cells.
Methods:
The whole gene sequence of circHERC4 was obtained from circBase database and cloned into cirRNA expression vector pLC5-ciR. The recombinant pLC5-ciR(+) hsa_circ_0007113 was constructed in vitro. The recombinant plasmid was identified by PCR, enzyme digestion and sequencing. The recombinant plasmid was transfected into HEK293 T and IMR90 cells with liposome 2000 transfection reagent. The expression of circHERC4 in normal cells, empty vector group and recombinant vector group was detected by RT-PCR, and the senescence of cells was detected by SA-β-Gal staining. Cell proliferation was detected by CCK-8 method.
Results:
The overexpression vector of circHERC4 was successfully constructed by correct sequencing, and circHERC4 could be efficiently transfected in HEK293 T cells. Compared with the control group, the positive rate of SA-β-Gal staining in the recombinant vector group decreased(P<0.05), and the proliferation rate of cells increased(P<0.05). Overexpression of circHERC4 could improve the proliferation and inhibit the senescence in IMR90 cells.
Conclusion
It suggests that circHERC4 has potential function of anti-senescence, which lays a foundation for further study on the function of circHERC4.