ABSTRACT
Objective:To investigate the effects of histone deacetylase 3 (HDAC3) on the pyroptosis of breast cancer (BC) cells via regulating miR-625/anti-silencing function 1B (ASF1B) and its mechanism.Methods:The expression level of HDAC3, miR-625 and ASF1B in BC tissue, adjacent normal tissue, BC cell lines (T47D, MCF7 and MDA-MB-231) and human normal breast epithelial cell MCF-10A was detected by qRT-PCR. The expression level of cell pyroptosis related protein NLRP3, Caspase-1 and GSDMD was detected by Western blot. The expression level of IL-18 and IL-1βwere detected by ELISA. ChIP experiment was used to determine the interaction between HDAC3 and miR-625. The dual luciferase reporter assay was used to verifiy the targeted regulation between miR-625 and ASF1B.Results:Compared with adjacent normal tissue and MCF-10A cells, the expression of HDAC3 and ASF1B was increased and the expression of miR-625 was decreased in BC tissue and cells (all P<0.05) . Compared with si-NC group, the protein expression level of NLRP3, Caspase-1 and GSDMD in si-HDAC3 group was increased, and the concentration of IL-18 and IL-1β in cell culture supernatant was increased (all P<0.05) . HDAC3 inhibited the expression of miR-625 by binding to the promoter region of miR-625 ( P<0.05) . Compared with si-HDAC3+miR-NC group, The expression of NLRP3, Caspase-1, GSDMD, IL-18 and IL-1β in si-HDAC3+miR-625 inhibitor group was decreased (all P<0.05) . ASF1B was confirmed as a target gene of miR-625, the level of pyroptosis related factors in si-HDAC3+pcDNA3.1-ASF1B group was significantly lower than that in si-HDAC3 + pcDNA3.1-NC group. Conclusion:HDAC3 up regulates the expression of ASF1B by inhibiting miR-625, and then inhibits BC cell pyroptosis.
ABSTRACT
@#[Abstract] Objective: To investigate the effects of miR-625 and Resistin on the proliferation, invasion and migration of NSCLC cells as well as the growth of transplanted tumors in nude mice and their possible mechanisms. Methods: qPCR was used to detect the expression of miR-625 and Resistin in 80 pairs of NSCLC and corresponding para-cancerous tissues (specimens collected from NSCLC patients who were surgically treated in Xinjiang Uygur Autonomous Region People’s Hospital from March 2018 to October 2019) and four cell lines. Bioinformatics was adopted to predict the targeting relationship between miR-625 and Resistin, which was then verified by Dual luciferase gene reporter experiment. Overexpression or inhibition of miR-625 and Resistin in NSCLC cells was achieved with lipofection transfection technology, and the experimental cells were divided into miR-625 mimic group, miR-625 inhibitor group, si-Resisitin group, miR-625 inhibitor+si-Resisitin group and NC group. The effects of miR-625 and Resistin on proliferation, invasion and migration of NSCLC cells were detected by CCK-8, Transwell and Scratch test, respectively. Western blotting was used to detect the effects of miR-625 and Resistin on the expressions of PI3K/AKT/Snail pathway proteins related with EMT in NSCLC cells. A549 cell transplanted tumor model was constructed in nude mice to observe the effect of miR-625 and Resistin on the growth of xenografts. Results: Compared with para-cancerous tissues, miR-625 showed low expression while Resistin showed high expression in NSCLC tissues and four cell lines (both P<0.01), and the two were negatively correlated (r=-0.7183,P<0.01). The expression of Resistin was related to the degree of NSCLC differentiation, clinical stage and lymph node metastasis. Resistin was a target gene of miR-625. Compared with the Blank group and NC group, the proliferation, invasion and migration of NSCLC cell linesA549 and H226, as well as the growth of transplanted tumors in nude mice in the miR-625 mimic group and the si-Resistin group were significantly reduced (all P<0.05), while those indicators in the miR-625 inhibitor group were significantly improved (all P<0.05); However, co-transfection of miR-625 inhibitor and si-Resistin significantly reversed the effect of miR-625 inhibitor on above indicators (all P<0.05);And there was no significant difference between NC group and miR-625 inhibitor+si-Resistin group (all P >0.05). The protein expressions of p-AKT, p-PI3K, Snail, Twist1 and Vimentin also showed the same trend (all P<0.05), while the expression of E-cadherin protein changed in the opposite direction (P<0.05). Conclusion: miR-625 is lowly expressed in NSCLC tissues and cell lines, which can negatively regulate Resistin expression to inhibit the proliferation, invasion and migration of NSCLC cells and the growth of transplanted tumors in nude mice. The mechanism may be related to the PI3K/AKT/Snail signaling pathway.
ABSTRACT
Objective: To analyze the correlation of miR-625 expression with clinicopathological characteristics in esophageal squa-mous cell carcinoma (ESCC) and to explore the effect of miR-625 on the migration and proliferation of ESCC cells. Methods:The expres-sion level of miR-625 was determined through real-time PCR in 86 paired human ESCC tissue specimens and tumor-adjacent normal esophageal tissue specimens, ESCC cell lines, and esophageal epithelial cell line. The associations of miR-625 expression with clinico-pathological characteristics and survival in ESCC patients were analyzed. Transwell and CCK-8 assays were performed to examine the effect of miR-625 expression on migration and proliferation of ESCC cells. Results:Compared with tumor-adjacent normal specimens, miR-625 was significantly downregulated in ESCC tissue specimens (P<0.05). MiR-625 expression was decreased in ESCC cell lines com-pared with human esophageal epithelial cell lines (P<0.05). Lower miR-625 expression was associated with poorer prognosis and sur-vival. The migration and proliferation abilities of ESCC cells were inhibited by miR-625 overexpression (P<0.05). Conclusion:MiR-625 acts as a tumor suppressor gene in the development and progression of ESCC, suggesting that miR-625 may serve as an efficient prog-nosis biomarker and a potential therapeutic target for ESCC.