Expression of miR-381 in tumor tissues and peripheral blood of HCC patients and the regulatory effect of its overexpression on polarization pattern of macrophages
WU Shuzhi
WANG Jing
Abstract：Objective To observe the expression changes of miR-381 in the tumor tissue,peripheral blood,and cul-tured macrophages of hepatocellular carcinoma(HCC)patients,and to explore the regulatory effect of miR-381 overexpres-sion on polarization pattern of macrophages.Methods In 40 HCC patients,tumor tissues and adjacent tissues were col-lected during operation,and the mRNA levels of miR-381,nitric oxide synthase(iNOS,M1 macrophage marker)and argi-nase 1(Arg-1,M2 macrophage marker)were detected by RT-PCR.The iNOS and Arg-1 proteins were detected by West-ern blotting.Peripheral venous blood of HCC patients and healthy volunteers was collected,and mRNA of miR-381,CD86(M1 macrophage marker)and CD163(M2 macrophage marker)were determined by RT-PCR,and proteins of CD86 and CD163 were determined by flow cytometry.Human myeloid leukemia monocyte THP-1 was induced to differentiate in-to M0 type macrophages(M0 group),M2 type macrophages(M2 group)and tumor-associated macrophages(TAMs,CM group),respectively.In each group,miR-381,iNOS mRNA,Arg-1 mRNA,CD86 mRNA,and CD163 mRNA were mea-sured by RT-PCR,and iNOS,Arg-1,CD86,and CD163 proteins were measured by Western blotting and flow cytometry.M0 macrophages were transfected with miR-381 mimic and mimic NC,respectively,and were labeled as the miR-381 mimic group and mimic NC group,and we established the miR-381 overexpression cell model.RT-PCR was used to detect miR-381,iNOS mRNA,Arg-1 mRNA,CD86 mRNA,and CD163 mRNA;Western blotting and flow cytometry were used to detect iNOS,Arg-1,CD86,and CD163 proteins.Results The relative expression levels of miR-381,iN-OS mRNA and protein in tumor tissues of HCC patients were lower than those in the adjacent tissues,while the relative ex-pression levels of Arg-1 mRNA and protein were higher than those in the adjacent tissues(all P<0.05).The expression levels of miR-381 and CD86 mRNA and protein in the peripheral blood macrophages of HCC patients were lower than those of healthy volunteers,while the expression levels of CD163 mRNA and protein were higher than those of healthy volunteers(all P<0.05).The relative expression levels of miR-381,iNOS,Arg-1,CD86,CD163 mRNA and protein in macro-phages of CM group was intermediate between M0 and M2 groups(all P<0.05).The mRNA and protein expression levels of miR-381,iNOS and CD86 in macrophages of the miR-381 mimic group increased,while the mRNA and protein expres-sion levels of Arg-1 and CD163 decreased(all P<0.05).Conclusions MiR-381 is low expressed in tumor tissues,pe-ripheral blood and cultured macrophages in vitro,and overexpression of miR-381 can convert macrophages from M2 type polarization to M1 type polarization.
Keywords：hepatocellular carcinomamiR-381macrophagestumor-associated macrophagesM1 type macro-phagesM2 type macrophages
Publication Date：2023-11-25
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 1-6 )