数字中医药（英文）2023，Vol.6Issue(1)：86-96,11.DOI:10.1016/j.dcmed.2023.02.008
益肾通癃汤通过上调miR-145-5p抑制TLR4/p38 MAPK/NF-κB信号通路抗前列腺癌作用机制研究
Mechanism of Yishen Tonglong Decoction inhibiting TLR4/p38 MAPK/NF-κB signaling pathway against prostate cancer via upregulating miR-145-5p
摘要
Abstract
Objective To investigate the mechanism of Yishen Tonglong Decoction(益肾通癃汤,YSTLD)inhibiting the toll-like receptor 4/p38 mitogen activated protein kinases/nuclear factor kappa-B(TLR4/p38 MAPK/NF-κB)signaling pathway against prostate cancer by up-regulat-ing miR-145-5p.Methods miRNA microarray technology was used to detect the changes of miRNA expres-sion profile in prostate cancer PC-3 cells treated with YSTLD,and miRNAs with marked differ-ences in miRNA microarray results were screened and validated by real-time polymerase chain reaction(qRT-PCR).Lentiviral transfection of miR-145-5p into prostate cancer PC-3 cells,Cell Counting Kit-8(CCK8)assay,and scratch assay were adopted to detect the effects of miR-145-5p on prostate cancer PC-3 cell proliferation and migration.qRT-PCR and Western blot were employed to detect the effects of miR-145-5p on TLR4/p38 MAPK/NF-κB signaling pathway and the expression levels of apoptosis-related genes caspase3,tumor necrosis factor-α(TNF-α),Bax,and Bcl-2.qRT-PCR and Western blot were used to detect the effects of serum containing YSTLD on miR-145-5p,TLR4/p38 MAPK/NF-κB signaling pathway,and the ex-pression levels of apoptosis-related genes caspase3,TNF-α,Bax,and Bcl-2.Results The expression levels of 35 miRNAs in prostate cancer PC-3 cells treated with YSTLD were significantly different from those in the control group,with miR-145-5p being the most significantly different;qRT-PCR validation revealed that the miR-145-5p levels in prostate cancer PC-3 cells treated with YSTLD were significantly higher than those in the DMSO con-trol group(P<0.05).After lentiviral transfection of miR-145-5p into prostate cancer PC-3 cells,miR-145-5p was found to inhibit the proliferation and migration of prostate cancer PC-3 cells.Overexpression of miR-145-5p up-regulated expression levels of caspase3,TNF-α,and Bax mRNA,and down-regulated expression levels of p38 MAPK,p65 NF-κB,and Bcl-2 mRNA in prostate cancer PC-3 cells(P<0.05),while up-regulated caspase3 protein expression levels in prostate cancer PC-3 cells and down-regulated expression levels of TLR4,p38 MAPK,and p65 NF-κB protein(P<0.05).Serum containing YSTLD could up-regulate the expression levels of caspase3,TNF-α,and Bax mRNA,and down-regulate the mRNA expression levels of p38 MAPK,p65 NF-κB,Bcl-2,and TNF receptor-associated factor 1(TRAF1)in prostate can-cer PC-3 cells after intervening prostate cancer PC-3 cells(P<0.05).Simultaneously,it up-regulated the expression levels of caspase3 protein and down-regulated the protein expression levels of TLR4,p38 MARK,p65 NF-κB,and TRAF1 in prostate cancer PC-3 cells(P<0.05).Conclusion YSTLD can promote apoptosis of prostate cancer PC-3 cells by up-regulating the expression level of miR-145-5p and inhibiting TLR4/p38 MAPK/NF-κB signaling pathway,which may be an important mechanism of YSTLD against prostate cancer.关键词
前列腺癌/益肾通癃汤/基因芯片技术/miR-145-5p/TLR4/p38MAPK/NF-κB信号通路Key words
Prostate cancer/Yishen Tonglong Decoction(益肾通癃汤,YSTLD)/Gene chip technology miR-145-5p/TLR4/p38 MAPK/NF-κB signaling path-way引用本文复制引用
涂雅玲,刘德果,羊羡,李博,陈其华..益肾通癃汤通过上调miR-145-5p抑制TLR4/p38 MAPK/NF-κB信号通路抗前列腺癌作用机制研究[J].数字中医药（英文）,2023,6(1):86-96,11.基金项目
Postgraduate Scientific Research Innovation Key Project of Hunan Province(CX20210680),Key Scientific Re-search Project of Hunan Provincial Department of Educa-tion(21A0224),Scientific Research Project of Hunan Health Commission(202204083613),and CHEN Qihua National Studio for Inheritance of TCM Famous Veteran Doctors(2022). （CX20210680）
