RESUMEN
Abstract Background: CTHRC1 is highly expressed in various cancers. Objectives: The aim of the study was to study the role of CTHRC1 played in lung adenocarcinoma (LUAD) development and its underlying biological functions. Methods: Enriched pathways and upstream transcription factors of CTHRC1 were explored by bioinformatics analysis. Dual-luciferase assay and Chromatin immunoprecipitation assay were used to verify the binding relationship between CTHRC1 and HOXB9. CCK-8 was utilized to detect cell viability. Expression levels of CTHRC1, HOXB9, and angiogenesis-related genes were assessed by quantitative real time-polymerase chain reaction. Angiogenesis assay was used to detect angiogenesis ability. Quantitative analysis of metabolites were used to detect the accumulation of neutral lipids, the levels of free fatty acids (FAs), and glycerol. Western blot was conducted to measure expression of metabolic enzymes of FA. Results: CTHRC1 was enriched in FA metabolic pathway, which was positively correlated and bound with HOXB9. CTHRC1 and HOXB9 expression was remarkably up-regulated in LUAD cells. Overexpression of CTHRC1 promoted FA metabolic pathway and angiogenesis, and FA inhibitor Orlistat restored it to NC group level. Meanwhile, CTHRC1 affected LUAD angiogenesis by activating HOXB9 to regulate FA metabolism. Conclusions: This study found that activation of CTHRC1 by HOXB9 induces angiogenesis by mediating FA metabolism. CTHRC1 may be a potential target for LUAD diagnosis.
RESUMEN
BACKGROUND: The aim of this study was to investigate the effect role and mechanism of miR-30b-3p on ovarian cancer cells biological function. METHODS: The expression of miR-30b-3p was detected in ovarian cancer cell lines and normal ovarian epithelial cell line by qRT-PCR. Mir-30b-3p mimic was transfected into OVCAR3 cells. Cell-counting kit-8 (CCK-8) assay was conducted to explore the effect of mir-30b-3p on the OVCAR3 cells' proliferation. Cell cycle and apoptosis were detected by Flow cytometry. Cell invasion ability was detected by Transwell test. The regulation of putative target of miR-30b-3p was verified by double luciferase reporter assays and Western blot. RESULT: We found that miR-30b-3p was downregulated in OVCAR3 cells. Overexpression of miR-30b-3p suppressed proliferation, promoted apoptosis, slowed cell cycle and inhibited migration and invasion of OVCAR3 cells. Bioinformatics analysis identified 3'-untranslated region (3'UTR) of Collagen triple helix repeat-containing 1 (CTHRC1) as the presumed binding site for miR-30b-3p. Detection of double luciferase reporter and Western-Blot result confirmed that CTHRC1 was the target gene of miR-30b-3p. Furthermore, E-cadherin, ß-cadherin and Vimentin protein expression level were changed after transfection of miR-30b-3p. CONCLUSION: miR-30b-3p function as an anti-cancer gene. Overexpression of miR-30b-3p can inhibit the biological function of ovarian cancer cells. MiR-30b-3p targets CTHRC1 gene plays an important role in epithelial-mesenchymal transformation (EMT), and supports miR-30b-3p as a potential biological indicator for ovarian cancer in the future.
Asunto(s)
Humanos , Femenino , Neoplasias Ováricas/genética , Regulación Neoplásica de la Expresión Génica/genética , Proteínas de la Matriz Extracelular/genética , MicroARNs/genética , Transición Epitelial-Mesenquimal/genética , Neoplasias Ováricas/metabolismo , Transducción de Señal , Movimiento Celular , Proteínas de la Matriz Extracelular/metabolismo , Apoptosis , Línea Celular Tumoral , Proliferación Celular , Invasividad NeoplásicaRESUMEN
Objective·To investigate the role of collagen triple helix repeat containing-1 (CTHRC1) in ovarian cancer cell metastasis and the related mechanism. Methods·The expression of CTHRC1 in ovarian cancer cells was detected by Western blotting. The cell line which had high expression of CTHRC1 was transfected with a CTHRC1 specific shRNA, and the lenti-CTHRC1 was used to overexpress CTHRC1 in the cell line whose expression of CTHRC1 was very low. Then the expression of Slug and MMP-2 was assessed. Transwell assay was used to determine the migration and invasion capability of ovarian cancer cells after transfection. Results·The expression of CTHRC1 in HO8910 cells was the lowest, while the CTHRC1 protein level was dramatically increased after transfection of lenti-CTHRC1. Meanwhile, there was a distinct rise of the migration and invasion ability, as well as the expression of Slug and MMP-2 (all P<0.05). Conversely, CaOV3 cells had a higher protein expression of CTHRC1. By using lenti-shCTHRC1, a remarkable knockdown of CTHRC1 was obtained. Likewise, the capability of migration and invasion was decreased, and the Slug and MMP-2 expression was reduced (all P<0.05). Conclusion·CTHRC1 might positively regulate Slug and MMP-2 to promote ovarian cancer cell metastasis.