KIAA1671-AS1/miR-588/STAT3 Axis Promotes Malignant Biological Behavior In Gastric Cancer Cells

Objective: Gastric cancer is one of the most common malignant tumors in China and the world,the fifth most common cancer,and the fourth leading cause of cancer death,with an average of 1 in 33 men and 1 in 78 women suffering from gastric cancer.In recent years,the incidence of gastric cancer in China and the world is still very serious.Based on the existing research on gastric cancer,it is still of great significance to further investigate the pathogenesis and potential treatment modalities of gastric cancer.Long non-coding RNAs do their jobs in the body by regulating the expression of various genes in the form of RNA molecules,and most do not encode any proteins themselves.KIAA1671-AS1 is an antisense lnc RNA in the lnc RNA family,located at position 22q11.23 of chromosome 22.The length of mature KIAA1671-AS1 is 973 nt.As a newly discovered lnc RNA,KIAA1671-AS1 has only been found to have a possible oncogenic effect in lung cancer,and its specific role and mechanism in gastric cancer are still unclear.The STAT3 gene,located on chromosome 17,is a key member of the STAT family of transcription factors and has been shown to have an oncogenic role in cancer and has been found to promote tumor development in gastric cancer.However,the application of STAT3 for cancer treatment is still in the preliminary stage and more possible targets still need to be found.This study aims to explore whether KIAA1671-AS1 plays a role in the development of gastric cancer through the miR-588/STAT3 axis and its specific mechanism,and to provide new ideas for the treatment of gastric cancer.Methods: In this study,the information of 375 gastric cancer tissues and 32 normal tissues adjacent to gastric cancer in TCGA was analyzed using the UALCAN website to investigate the expression level of KIAA1671-AS1 in gastric cancer,which was further verified by q RT-PCR assay in gastric cancer cells.In this study,MKN-45 cell line with high expression of KIAA1671-AS1 and HGC-27 cell line with low expression of KIAA1671-AS1 were selected for cell transfection to regulate the expression level of KIAA1671-AS1,and the proliferation level,migration ability and invasiveness of gastric cancer cells were assessed by CCK8 assay,cell scratch healing assay and Transwell cell invasion assay.Using the GEPIA2 website,STAT3 was found to be abnormally highly expressed in gastric cancer and positively correlated with KIAA1671-AS1.q RT-PCR and Western blot assays were performed to verify the expression level of STAT3 in gastric cancer cells and the positive regulation of STAT3 by KIAA1671-AS1,KIAA1671-AS1 si RNA and STAT3 overexpression plasmid cotransfection rescue assay confirmed the role of KIAA1671-AS1 in gastric cancer cells through STAT3.Bioinformatics software was used to analyze and explore the competitive binding of miR-383-5p and miR-588-5p by KIAA1671-AS1 and STAT3 through ce RNA mechanism.miR-383-5p and miR-588-5p were detected by q RT-PCR in various gastric cancer cell lines and gastric cancer cells with different KIAA1671-AS1 expression levels The expression levels of KIAA1671-AS1 in each gastric cancer cell line and gastric cancer cells with different KIAA1671-AS1 expression levels were further investigated to investigate the regulation of STAT3 expression by miR-383-5p and miR-588-5p.The potential sites of binding between the promoter region of KIAA1671-AS1 and the transcription factor STAT3 were found through online databases such as h TFtarget and JASPAR.q RT-PCR experiments were performed to detect the expression level of KIAA1671-AS1 by regulating the expression level of STAT3 to explore the feedback effect of STAT3 on KIAA1671-AS1.Results: 1.KIAA1671-AS1 was highly expressed in gastric cancer and correlated with gender.KIAA1671-AS1 expression level was regulated by cell transfection in gastric cancer cells and cell biological behavior assay was performed.2.STAT3 expression in gastric cancer was upregulated and positively correlated with KIAA1671-AS1,and the expression level of KIAA1671-AS1 was regulated by cell transfection.q RT-PCR and Western blot experiments revealed that KIAA1671-AS1 positively regulated STAT3 expression in gastric cancer cells,and the results of the rescue experiments further suggested that KIAA1671-AS1 promoted gastric cancer cells by regulating STAT3 to promote malignant biological behavior in gastric cancer cells.miR-383-5p and miR-588-5p were shown to be potential binding sites for KIAA1671-AS1 and STAT33’UTR in online databases such as RNA22 and star Base.q RT-PCR experiments on different gastric cancer cell lines and gastric cancer cells with different KIAA1671-AS1 expression levels The results of q RT-PCR experiments verified the negative regulation of miR-383-5p and miR-588-5p by KIAA1671-AS1,suggesting that KIAA1671-AS1 can play a role in gastric cancer cells by inhibiting miR-383-5p and miR-588-5p and thus upregulating STAT3 expression.Transcription factor databases such as JASPAR and h TFtarget suggested the existence of potential binding sites for STAT3 in the promoter region of KIAA1671-AS1 gene.q RT-PCR results indicated that the expression of KIAA1671-AS1 increased with the overexpression of STAT3.Conclusion: 1.KIAA1671-AS1 was aberrantly highly expressed in gastric cancer,correlated with gender,and was able to enhance the proliferation,migration and invasion of gastric cancer cells and promote malignant biological behavior of gastric cancer cells.2.KIAA1671-AS1 promoted malignant biological behavior of gastric cancer cells by regulating transcription factor STAT3 expression in gastric cancer cells and promoted malignant biological behavior of gastric cancer cells by inhibiting miR-383-5p and miR588-5p upregulated STAT3 expression,and STAT3 also had a positive feedback effect on KIAA1671-AS1.