临床与病理杂志2023，Vol.43Issue(9)：1621-1630,10.DOI:10.11817/j.issn.2095-6959.2023.220033
MiR-148b-3p/FOXO4轴对乳腺癌细胞自噬及生长的影响
Effects of miR-148b-3p/FOXO4 axis on autophagy and growth of breast cancer cells
摘要
Abstract
Objective:The role of miR-148b-3p in breast cancer(BC)progression through the regulation of forkhead box O4(FOXO4)remains incompletely elucidated.This study investigates the effects of miR-148b-3p-mediated regulation of FOXO4 expression on BC cell proliferation,apoptosis,and autophagy. Methods:The expression of miR-148b-3p and FOXO4 in BC patient tumor specimens(BC tissues)and adjacent non-tumor specimens(normal tissues),as well as in normal human breast epithelial cells(MCF-10A)and human BC cells(MCF7,SKBR3,MDA-MB-231 cells)were assessed by real-time reverse transcription polymerase chain reaction(real-time RT-PCR).MDA-MB-231 cells were transfected and divided into a control group,a inhibitor-NC group,a miR-148b-3p inhibitor group,a si-NC group,a si-FOXO4 group,a miR-148b-3p inhibitor+si-NC group,and a miR-148b-3p inhibitor+si-FOXO4 group.Cell proliferation was detected by cell counting kit 8(CCK-8)assay.Apoptosis was detected by flow cytometry.Autophagy was observed by transmission electron microscope.The expressions of FOXO4,proliferation,apoptosis,and autophagy-related proteins were detected by Western blotting.Dual luciferase reporter gene assay and miRNA pull-down assay were used to verify the targeting relationship between miR-148b-3p and FOXO4. Results:Compared to normal tissues,miR-148b-3p expression was upregulated,and FOXO4 mRNA expression was downregulated in BC tissues(P<0.05).Pearson correlation analysis revealed a negative correlation between miR-148b-3p and FOXO4 mRNA levels in BC patient tumor tissues(r=−0.774,P<0.01).Compared to MCF-10A cells,miR-148b-3p expression increased,while FOXO4 protein and mRNA expression decreased in MCF-7,SKBR3,and MDA-MB-231 cells(P<0.05),with MDA-MB-231 cells showing more significant changes.MiR-148b-3p negatively regulated the expression of FOXO4.Knocking down miR-148b-3p in MDA-MB-231 cells reduced cell viability and proliferating cell nuclear antigen(PCNA)protein levels,increased apoptosis rate,autophagosome number,FOXO4 mRNA and protein levels,as well as cleaved-caspase-3,cleaved-PARP-1,LC3-II/LC3-I,and Beclin-1 protein levels(P<0.05).Knocking down FOXO4 reversed these effects.Knockdown of FOXO4 attenuated the effects of knockdown of miR-148b-3p on MDA-MB-231 cell proliferation,apoptosis,and autophagy. Conclusion:MiR-148b-3p may promote MDA-MB-231 cell proliferation,inhibit apoptosis and autophagy by targeting FOXO4.关键词
miR-148b-3p/叉头框蛋白O4/乳腺癌/增殖/凋亡/自噬Key words
miR-148b-3p/forkhead box O4/breast cancer/proliferation/apoptosis/autophagy引用本文复制引用
郭玲,陈敬信..MiR-148b-3p/FOXO4轴对乳腺癌细胞自噬及生长的影响[J].临床与病理杂志,2023,43(9):1621-1630,10.基金项目
陕西省教育厅专项科研计划项目(17JK0209).This work was supported by the Special Research Project of Shaanxi Provincial Department of Education,China(17JK0209). （17JK0209）
