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Objective: To investigate the role of miR-129-5p in regulating HMGB1 expression in pancreatic cancer cell apoptosis. Methods: The untreated pancreatic cancer SW1990 cells were used as the control group. Mimics-NC (empty vector), miR-129-5p mimics, inhibitor-NC (empty vector)and miR-129-5p inhibitor were transfected into SW1990 cells by liposome transfection method as the mimics-NC group, miR-129-5p overexpression group (miR-129-5p mimics group), inhibitor-NC group and miR-129-5p low expression group (miR-129-5p inhibitor group). The binding site of miR-129-5p and HMGB1 was predicted by online target gene prediction website Target genes, and the targeting relationship between miR-129-5p and HMGB1 was verified by dual luciferase gene report experiment. The expression of miR-129-5p in each group was detected by qRT-PCR, and the expression of HMGB1 protein and apoptosis-related proteins Caspase 3 and Bcl-2 by Western blot. Hoechst staining was used to observe the changes of apoptosis. Results: Compared with the mimics-NC group and control group, miR-129-5p mimics transfection significantly up-regulated miR-129-5p level (P<0.01), inhibited HMGB1(P<0.01)and Bcl-2(P<0.05)protein expression, pro-moted Caspase 3 protein expression (P<0.05), and promoted apoptosis; compared with the inhibitor-NC group and control group, miR-129-5p inhibitor transfection significantly down-regulated miR-129-5p level (P<0.05), promoted HMGB1 and Bcl-2 protein expression (all P<0.05), inhibited Caspase 3protein expression (P<0.01), and inhibited apoptosis. The results of dual luciferase reporter gene assay showed that miR-129-5p could inhibit the fluorescence activity of wildtype HMGB1 cells and target the expression of HMGB1. Conclusion: miR-129-5p promotes the apoptosis of pancreatic cancer SW1990 cells by targeting inhibition of HMGB1 expression. [ABSTRACT FROM AUTHOR]

Background: Glucose 6 phosphatase dehydrogenase (G6PD) is a key regulator of the pentose phosphate pathway (PPP). However, the exact role of G6PD in gastrointestinal cancers remains unclear. The purpose of this study is to explore the correlation of G6PD with clinical features, pathological stages, diagnosis and prognosis of gastrointestinal cancers, as well as uncover possible mechanisms of G6PD on mutations, immunity and signaling pathways., Methods: G6PD mRNA expression data were downloaded from TCGA and GEO databases. Protein expression was examined by the HPA database. The correlation of G6PD expression with clinical and pathological characteristics was explored. The pROC package in R language was used to evaluate the diagnostic value of G6PD expression in gastrointestinal cancers. We accessed the correlation of disease-free survival (DFS) with G6PD online by Kaplan-Meier plotter. Univariate Cox regression and stepwise multiple Cox regression analysis were performed to determine the association between G6PD and patient's overall survival. In addition, genomic alterations, mutation profiles, immune infiltration, drug sensitivity and enrichment analysis related with G6PD were visualized., Results: After a pan-cancerous genomic analysis, we found that G6PD expression was the highest in African American esophageal carcinoma (ESCA) patients ( P <0.05). G6PD was correlated with age, weight, disease stage, lymph node metastasis and pathological grade. Notably, G6PD showed an excellent predictive diagnosis ability for liver hepatocellular carcinoma (LIHC) (AUC=0.949, 95% CI=0.925-0.973, P <0.001). G6PD can improve the DFS of esophageal adenocarcinoma (EAC) and pancreatic adenocarcinoma (PAAD) patients ( P <0.05). Both Univariate Cox regression and stepwise multiple Cox regression analysis in R language determined that G6PD expression was closely related with LIHC ( P <0.001). G6PD was found to have a high mutation rate in colon adenocarcinoma and ESCA and gene amplification in ESCA, Cholangiocarcinoma, PAAD and LIHC. Copy number of G6PD was missing in LIHC. G6PD was also related to mutation of TP53 ( P <0.05). Particularly, it was positively correlated with CD276 in all gastrointestinal cancers and negatively with HERV-H LTR-associating 2 in ESCA and stomach adenocarcinoma. The abnormal expression of G6PD was related to the increase of CD4+ Th2 subsets and the decrease of CD4+ (non-regulatory) of T cells. G6PD was sensitive to FK866, Phenformin, AICAR etc., while resistant to RO-3306, CGP-082996, TGX221 etc. G6PD was found to closely interact with TALDO1, GAPDH and TP53. G6PD related biological processes included aging, nutritional response and daunorubicin metabolism, and related pathways included PPP, cytochrome P450 metabolism of exogenous substances and glutathione metabolism., Conclusion: G6PD is highly expressed in gastrointestinal cancers. It is a carcinogenic indicator related to prognosis and can be used as a potential diagnostic marker of gastrointestinal cancers, so as to provide new strategy for cancer treatment., Competing Interests: None., (AJTR Copyright © 2023.)