摘要
目的:探究miR-122-5p在婴儿胆道闭锁(BA)中的作用及其机制。方法:RT-qPCR检测miR-122-5p在BA患儿及胆总管囊肿(CC)患儿血清中的表达,及miR-122-5p和PPP2R2A mRNA在LX-2细胞中的表达。CCK-8检测不同组别中LX-2细胞的增殖活性。Western blot检测p-AKT、AKT、PPP2R2A及Ki67蛋白在LX-2细胞内的表达。通过生物信息学网站预测miR-122-5p的下游靶基因,并通过双荧光素酶报告基因实验验证其靶向关系。结果:miR-122-5p在BA患儿血清中的表达水平显著升高。抑制miR-122-5p的表达显著抑制LX-2细胞的增殖、细胞内AKT的激活并促进LX-2细胞的凋亡,过表达PPP2R2A对LX-2细胞生物学行为的影响与前述一致。双荧光素酶报告基因实验证实PPP2R2A是miR-122-5p的下游靶基因。同时过表达LX-2细胞内miR-122-5p及PPP2R2A的表达水平对LX-2细胞的生物学行为无显著影响。结论:miR-122-5p在BA患儿血清中过表达,并通过体外实验证实miR-122-5p靶向PPP2R2A影响AKT信号通路的激活调控肝纤维化的进程。
Objective:To explore the role and mechanism of miR-122-5p in BA.Methods:RT-qPCR was used to detect the expression of miR-122-5p in the serum of BA and CC children,and the expression of miR-122-5p and PPP2R2A mRNA in LX-2 cells.The proliferation activity of LX-2 cells in different groups was detected by CCK-8.Western blot was used to detect the expression of p-AKT,AKT,PPP2R2A,and Ki67 protein in LX-2 cells.The downstream target genes of miR-122-5p were predicted through bioinformatics websites,and their targeting relationships were verified by dual-luciferase reporter gene experiments.Results:The expression level of miR-122-5p in the serum of BA children increased significantly.Inhibiting the expression of miR-122-5p significantly inhibited the proliferation of LX-2 cells,the activation of intracellular AKT,and promoted the apoptosis of LX-2 cells.The effect of over expression of PPP2R2A on the biological behavior of LX-2 cells was consistent with that described above.The dual-luciferase reporter gene experiment confirmed that PPP2R2A was the downstream target gene of miR-122-5p.At the same time,the expression levels of miR-122-5p and PPP2R2A in LX-2 cells had no significant effect on the biological behavior of LX-2 cells.Conclusion:miR-122-5p is overexpressed in the serum of BA children,and in vitro experiments confirmed that miR-122-5p targeting PPP2R2A affects the activation of the AKT signaling pathway and regulates the process of liver fibrosis.
作者
赵成基
李斌德
李刚
王文赟
马仲福
曾永娟
胡继科
ZHAO Cheng-Ji;LI Bin-De;LI Gang;WANG Wen-Yun;MA Zhong-Fu;ZENG Yong-Juan;HU Ji-Ke(Department of Pediatric Surgery,Second Hospital of Lanzhou University,Lanzhou 730000,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2020年第16期1998-2002,共5页
Chinese Journal of Immunology
基金
甘肃省自然科学基金(1606RJZA102)资助。
