| Background:Primary liver cancer is one of the most common malignancies and the third leading cause of cancer-related death worldwide.Hepatocellular carcinoma(HCC)is the most common type of liver cancer and is treated with surgery,ablation,intervention,chemotherapy and targeted therapy.Since most HCC patients are diagnosed at an advanced stage,the prognosis of HCC is poor at present.Therefore,it is of great significance to explore the mechanism of HCC occurrence and development as well as diagnostic markers and therapeutic targets for HCC.Long noncodingRNAs(LncRNAs)are a kind of non-codingRNAs,and m6 A is the most commonRNA modification mode.The interaction between LncRNA and m6 A modification plays an important role in cancer.SPATA41-202 is a novel LncRNA with low expression and high m6 A methylation in HCC,and its role in HCC remains unclear.Objective: The purpose of this study was to investigate the correlation between the interaction between SPATA41-202 and m6 A modification and the proliferation,invasion and metastasis of HCC,and to explore the potential mechanism and clinical significance.Method:1.Human m6A-lncRNA Epitranscriptomic Microarray detection was performed on 3 paired samples of cancer and adjacent tissues of patients with hepatocellular carcinoma,and verified in tissues and cells of HCC.It was found that SPATA41-202 had high methylation level of m6 A and low expression in HCC.Surgical samples and clinicopathological characteristics of 100 HCC patients were randomly collected.The expression of SPATA41-202 in HCC and adjacent tissues was detected by RT-q PCR,and the relationship between the expression of SPATA41-202 and clinicopathological characteristics was further analyzed.2.In vitro experiment: SPATA41-202 was overexpressed in LM3 and 97H cells and knocked down in HEPG2 and 97 L cells.The cell proliferative activity of different treated cells was detected by CCK8,plate cloning and Ed U experiment,and the apoptosis of cells was evaluated by flow cytometry.Transwell migration and Matrigel invasion assay were used to evaluate cell migration and invasion.3.In vivo experiment: SPATA41-202 was overexpressed in LM3 cells,and knocked down in HEPG2 cells.The cells were injected subcutaneously into mice to construct a mouse subcutaneous tumor model.Tumor formation was observed periodically and tumor size was measured to study the effect of SPATA41-202 on the growth of hepatocellular carcinoma in vivo.In order to study the effect of SPATA41-202 on hepatocellular carcinoma metastasis in vivo,a metastasis model was constructed by injecting above cells into the caudal vein,and fluorescence signal intensity of metastatic tumor was detected in vivo.HE and immunohistochemical staining were performed on subcutaneous tumor tissues,to detect the expression of Ki67,MMP-9,and cleaved Caspase-3.4.Bioinformatics method was used to predict the downstream target genes of SPATA41-202 and hsa-miR-27b-3p,fluorescence in situ hybridization,double luciferin reporter gene and rescue experiment for verification.The binding of SPATA41-202 to YTHDC2 protein was detected byRNA pull down assay and mass spectrometry.Me RIP-q PCR,RNA stability experiment and rescue experiment verified that YTHDC2 affected the stability of SPATA41-202 in an m6a-dependent manner.Results:1.SPATA41-202 has a high methylation level of m6 A and a low expression level in hepatocellular carcinoma.2.SPATA41-202 inhibits proliferation,invasion and migration of hepatocellular carcinoma in vitro,and promotes apoptosis.3.SPATA41-202 inhibits the growth and metastasis of hepatocellular carcinoma in vivo;4.SPATA41-202 regulates TNPO1 by competitively binding hsa-miR-27b-3p;5.SPATA41-202 affects Wnt pathway regulation of HCC and epithelial mesenchymal transformation through ceRNA mechanism;6.SPATA41-202 affects proliferation,apoptosis,invasion and metastasis of HCC through ceRNA mechanism;7.YTHDC2 affects the stability of SPATA41-202 in a m6 A dependent manner.Conclusions: YTHDC2 affects the stability of SPATA41-202 in an m6A-dependent manner,and SPATA41-202 affects the proliferation,invasion and metastasis of HCC by regulating Wnt pathway through ceRNA mechanism.Figures:14,Forms:30,Text references:90,Review references:210... |
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