lncRNA PCBP1-AS1 Aggravates the EMT of Hepatocellular Carcinoma via Regulating PCBP1/miR-199/XBP1 Pathway

HE Bin; HU Guohuang; XIA Dan

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Journal of Hunan Normal University(Medical Science) ›› 2023, Vol. 20 ›› Issue (6) : 35-43.

Basic Medicine

lncRNA PCBP1-AS1 Aggravates the EMT of Hepatocellular Carcinoma via Regulating PCBP1/miR-199/XBP1 Pathway

HE Bin, HU Guohuang, XIA Dan

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Abstract

Objective This study aims to investigate the regulatory role of lncRNA PCBP1-AS1 in EMT transformation of hepatocellular carcinoma and its possible mechanism, so as to provide a new target for the prevention and treatment of hepatocellular carcinoma. Methods Real-Time PCR was used to detect the expression of lncRNA PCBP1-AS1, miR-199, XBP1 and other genes in liver tissue of HCC patients or HCC cells. Pearson correlation coefficient was used to analyze the correlation between lncRNA PCBP1-AS1 expression and TNM stage of hepatocellular carcinoma. Bioinformatics was used for predicting the interaction between lncRNA PCBP1-AS1 and miR-199, which was verified via luciferase reporter gene experiment. Lipo2000 was used for transfering PCBP1-AS1 plasmid or miR-199 mimc (inhibitor) into Hep3B cells. Transwell assay was used for detecting the invasive ability of cells. Scratch test was used for detecting the migration ability of cells. CCK-8 kit was used for detecting the cell activity. LDH release assay was used for detecting the cytotoxicity. Western blot was used for detecting the expression of XBP1 and other genes. Results Compared with the normal liver tissues, the expression level of LncRNA PCBP1-AS1 in liver cancer tissues was significantlyincreased. The expression level of LncRNA PCBP1-AS1 was positively correlated with the TNM stage of hepatocellular carcinoma. It was predicted that lncRNA PCBP1-AS1 interacted with miR-199 mimic, and miR-199 mimic could significantly reduce the luciferase activity of Hep3B cells transfected with lncRNA PCBP1-AS1-wt luciferase reporter gene. LncRNA PCBP1-AS1 overexpression of can significantly increase the expression of lncRNA PCBP1-AS1, XBP1, Snai1 and Vimentin in Hep3B cells, significantly decrease the expression of miR-199 and E-cadherin, significantly increase the viability of H cells, and significantly reduce LDH release, significantly increase the migration and invasion ability of Hep3B cells, and these effects could be inhibited by miR-199 mimic or promoted by miR-199 inhibitor. Conclusion LncRNA PCBP1-AS1 can upregulatethe expression of Snai1 and Vimentin, downregulate the expression of E-cadherin in liver cancer cells and promote its epithelial-mesenchymal transformation (EMT) via sponging the expression of miR-199. LncRNA PCBP1-AS1 is a potential target for hepatocellular carcinoma.

Key words

hepatocellular carcinoma / lncRNA PCBP1-AS1 / miR-199 / XBP1 / epithelial mesenchymal transformation

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HE Bin, HU Guohuang, XIA Dan. lncRNA PCBP1-AS1 Aggravates the EMT of Hepatocellular Carcinoma via Regulating PCBP1/miR-199/XBP1 Pathway[J]. Journal of Hunan Normal University(Medical Science). 2023, 20(6): 35-43

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