Effect of micro ribonucleic acid-375 targeting phosphatidylinositol 3-kinase/protein kinase B pathway on high glucose-induced proliferation and angio-genesis of human retinal endothelial cells / 眼科新进展

Objective To investigate the influencing mechanism of micro ribonucleic acid(miR)-375 targeting phos-phatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)pathway on high glucose-induced proliferation and angiogenesis in human retinal endothelial cells(hRECs).Methods The hRECs were cultured in vitro,and transfection and dual lucifer-ase assay were performed on them.These hRECs were divided into the control group,high glucose group,high glucose+miR-375 group,and high glucose+miR-375+LM22B-10 group.The Cell Counting Kit-8 was used to detect the cell prolifera-tion ability,the angiogenesis assay was used to detect the vascular formation ability,real-time quantitative PCR was used to detect the miR-375 and PI3K mRNA expressions in hRECs,and Western blot was used to detect the PI3K and p-AKT/AKT protein expressions in hRECs.Results At 48 h and 72 h after the cultivation,compared with the control group,the pro-liferation viability,PI3K and p-AKT/AKT protein expressions,vascular formation ability,and PI3K mRNA expression in hRECs significantly increased,and the miR-375 expression in hRECs significantly decreased in the high glucose group,high glucose+miR-375 group and high glucose+miR-375+LM22B-10 group(all P＜0.05).Compared with the high glucose group,the proliferation viability,PI3K mRNA and protein expressions,p-AKT/AKT protein expression and vascular forma-tion ability in hRECs were significantly reduced,and miR-375 expression significantly increased in the high glucose+miR-375 group(all P＜0.05).Compared with the high glucose+miR-375 group,the proliferation viability,vascular formation a-bility and p-AKT/AKT protein expression in hRECs significantly increased in the high glucose+miR-375+LM22B-10 group(all P＜0.05);there was no significant difference in the miR-375 and PI3K mRNA(all P＞0.05).After transfected with miR-375 mimic and wt-PI3K-pGL4,the relative luciferase activity in hRECs significantly decreased compared with transfec-tion with miR-375 NC and mut-PI3K-pGL4(all P＜0.05).Conclusion The targeted inhibition of the PI3K/AKT pathway by miR-375 can suppress the high glucose-induced proliferation and angiogenesis of hRECs,alleviating DR.

RASSF1A expression mediated by lentivirus inhibits growth of small-cell lung cancer cell line H446 / 中国病理生理杂志

AIM: To explore the inhibitory effect of Ras-association domain family 1A ( RASSF1A) on the small-cell lung cancer cell growth .METHODS:The lentiviral expression vector containing RASSF1A gene was constructed and used to infect the small-cell lung cell line H446.The growth curve and cell cycle were detected by MTT assay and flow cytometry.The mRNA and protein levels of cell cycle-associated proteins were determined by real-time PCR and Western blotting.RESULTS:We obtained the H446 cells in which RASSF1A was stably expressed (named RASSF1A-H446). Compared with normal cell group and negative cell group , RASSF1A inhibited the proliferation of H446 cells, and arrested H446 cells in G1 phase.The expression of p21 and p27 was significantly increased , and E2F1 was significantly decreased in RASSF1A-H446 cells.CONCLUSION:RASSF1A inhibits the H446 cell growth by increasing the expressions of p 21 and p27, and decreasing the expression of E 2F1.