| miRNA is a type of non-coding single-stranded RNA molecule with a length of 22 nucleotides widely existing in the organism.It mainly regulates the expression of target genes by participating in post-transcriptional control of genes.miRNA can regulate various key genes related to cell growth,development,and differentiation,and is involved in various human diseases.Many studies have shown that miRNAs are closely related to tumor initiation,progression and invasion.miRNA can regulate cell cycle,apoptosis,cell migration,and tumor angiogenesis.Therefore,miRNA dysregulation in vivo is the key to the pathogenesis of many diseases including tumors.miR-16-5p is highly conserved among various species,is an important regulatory element in many key signaling pathways in the cell,and closely related to physiological processes such as cell cycle,apoptosis,and cell proliferation.According to reports,miR-16-5p is frequently deleted or downregulated in many types of common human cancer,and is related to the occurrence and development of cancer.However,little is known about its regulatory mechanism on hepatic carcinoma,and the detailed molecular mechanism has not been elucidated.Due to a tumor regulation role of miR-16-5p,this subject explore the effects and molecular mechanism of miR-16-5p on HepG2 in vitro,which may provide an important theoretical basis for the development of new therapies for liver cancer.First of all,in order to explore the possible role of miR-16-5p in hepatic carcinoma,we used q RT-PCR to detect the expression level of miR-16-5p in four different cell lines,and found that the expression level of miR-16-5p in three liver cancer cells was significantly lower than that of human normal hepatocyte 7702,so it was speculated that miR-16-5p might play a potential role in hepatic carcinoma.and the low expression in HepG2 cells was more significant,so human liver cancer cell line HepG2 cells were selected as experimental materials.Subsequently,we artificially interfered the expression of miR-16-5p in cells by transfecting its mimics or inhibitors.In addition,CCK-8 analysis,Ed U test,flow cytometry,wound healing analysis and Western Blot were used to detect cell viability,cell proliferation,cell cycle and cell migration ability when miR-16-5p level changes in HepG2.The results showed that overexpressed miR-16-5p significantly reduced the cell viability of HepG2 cells,suppressed the expression of cell proliferation and cycle-related genes,thereby inhibiting cell proliferation and inhibiting migration of HepG2 cells.it has been proved that Smad3 is the main intracellular receptor in the TGF-β signaling pathway,and promotes the cell proliferation and cell migration of invasive and metastatic cells in various cancers,such as lung cancer,prostate cancer and pancreatic cancer,and Smad3 has been identified as one of the downstream targets of miR-16-5p in various cancer tissues except hepatic carcinoma.To further verify the regulatory effect of the miR-16-5p/Smad3 pathway in HepG2 cells,Western Blot results further showed that Smad3 is a downstream target of miR-16-5p in HepG2 cells,and miR-16-5p can inhibit the expression of Smad3 protein at the post-transcriptional level,thereby regulating HepG2 cell proliferation.These results provide a theoretical basis for targeted therapy of hepatic carcinoma. |
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