ABSTRACT
Autologous chondrocyte (CH) transplantation is a novel strategy to treat post-traumatic osteoarthritis (PTOA). In this study, an in vitro coculture model was used to explore the effects of interleukin (IL)-10 overexpressed CHs on degenerated CHs. The original CHs were isolated from the patients' knee joint cartilage and pretreated with IL-1ß to get degenerated CHs. Moreoer, CHs were transfected with a lentivirus vector to overexpress IL-10. After coculture with the degenerated CHs, the apoptosis, collagen X, IL-6, and TNF-α of original CHs were increased, and the collagen II and IL-10 were decreased compared to the separated culture condition. Coculture with original CHs did not alleviate the degeneration of the IL-1ß-pretreated CHs. However, coculture with the IL-10-overexpressed CHs rescued the proliferation, collagen II, aggrecan, SOX9, and IL-10 expression, and suppressed the apoptosis, collagen X, RUnx2, IL-6, and TNF-α levels in the IL-1ß pretreated CHs. Additionally, the IL-10-overexpressed CHs also maintained a healthy state when cocultured with the degenerated CHs. Therefore, transplanting the IL-10-overexpressed CHs in the treatment of PTOA would obtain a more durable and visible effect in alleviating the CH degeneration.
Subject(s)
Chondrocytes , Osteoarthritis , Cell- and Tissue-Based Therapy , Cells, Cultured , Coculture Techniques , Humans , Interleukin-10/genetics , Interleukin-1beta/genetics , Osteoarthritis/therapyABSTRACT
OBJECTIVE: Colorectal cancer (CRC) is the fourth leading cause of death worldwide and there is a need for more specific therapeutic targets and biomarkers for the disease. Transforming growth factor ß1-induced transcript 1 (TGFΒ1I1) was reported to be downregulated in CRC tissues; however, the precise roles of TGFΒ1I1 in CRC remain unclear. PATIENTS AND METHODS: The expression of TGFΒ1I1 in CRC cell lines and tissues was assessed by quantitative Polymerase Chain Reaction (qPCR). TGFΒ1I1 was overexpressed in SW620 and RKO cells. Cell viability was analyzed by a CCK-8 assay. The proportion of apoptotic cells was analyzed by flow cytometry. The EdU cell proliferation assay of SW620 and RKO cells after transfection was performed via flow cytometry. The migration potency of SW620 and RKO cells was analyzed using a cell migration assay. A wound healing assay was performed to assess the migration potency of SW620 and RKO cells. The invasion potency of SW620 and RKO cells after TGFΒ1I1 overexpression was analyzed. The protein levels of VEGF, TGF-ß, MMP9, p-Smad2/3, N-cadherin, and E-cadherin were analyzed by Western blot. RESULTS: Decreased expression of TGFΒ1I1 was found in CRC tissues and cell lines. Overexpression of TGFΒ1I1 inhibited the proliferation and induced the apoptosis of CRC cells. The overexpression of TGFΒ1I1 inhibited the migration and invasion of CRC cells. We also found that the overexpression of TGFΒ1I1 in CRC cells inhibited the TGF-ß pathway and epithelial-mesenchymal transition (EMT) progress. CONCLUSIONS: TGFΒ1I1 suppressed cell migration and invasion in CRC by inhibiting the TGF-ß pathway and EMT progress.
Subject(s)
Cell Movement , Colorectal Neoplasms/metabolism , Epithelial-Mesenchymal Transition , Intracellular Signaling Peptides and Proteins/metabolism , LIM Domain Proteins/metabolism , Apoptosis , Cell Proliferation , Colorectal Neoplasms/genetics , Colorectal Neoplasms/pathology , Gene Expression Regulation, Neoplastic , HCT116 Cells , HT29 Cells , Humans , Intracellular Signaling Peptides and Proteins/genetics , LIM Domain Proteins/genetics , Neoplasm Invasiveness , Signal TransductionABSTRACT
OBJECTIVE: MicroRNA-338-3p (miR-338-3p) was reported to influence the metastasis and development of several human cancers. However, in bladder cancer (BC), the special function of miR-338-3p remains unknown. Here, we aimed at exploring the miR-338-3p function in the progression of BC. PATIENTS AND METHODS: miR-338-3p and ETS1 expressions were examined by quantitative Real-time polymerase chain reaction (qRT-PCR) in BC samples. Following that, transwell assays for cell migration and invasion were performed. And MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay for cell proliferation was conducted as well. Western blot was employed to examine the epithelial-mesenchymal transition (EMT) marker expressions. Finally, the relationship between miR-338-3p and E26 transformation specific-1 (ETS1) was verified by luciferase reporter assay. RESULTS: The decreased miR-338-3p expression was examined in BC cells. Moreover, miR-338-3p upregulation repressed cell proliferation ability in BC. Next, miR-338-3p upregulation also depressed cell metastasis and EMT in BC cells. Furthermore, ETS1 was a direct target of miR-338-3p and inversely associated with its expression. And upregulation of ETS1 partially rescued the suppression of miR-338-3p for cell proliferation and metastasis in BC. CONCLUSIONS: Upregulation of miR-338-3p inhibited the proliferation, metastasis and EMT in BC by suppressing ETS, showing that miR-338-3p might block the development of BC through regulating ETS1 expression.
Subject(s)
Cell Proliferation/genetics , Gene Expression Regulation, Neoplastic , MicroRNAs/genetics , Proto-Oncogene Protein c-ets-1/genetics , Urinary Bladder Neoplasms/genetics , 3' Untranslated Regions , Cell Line, Tumor , Cell Movement/genetics , Disease Progression , Epithelial-Mesenchymal Transition/genetics , Humans , Up-Regulation , Urinary Bladder Neoplasms/metabolism , Urinary Bladder Neoplasms/pathologyABSTRACT
OBJECTIVE: MicroRNAs (miRNAs) play critical roles in regulating tumor development and progression. The aim of the study is to investigate the clinical significance of miR-1294 expression in gastric cancer (GC). PATIENTS AND METHODS: The expression of miR-1294 in 82 cases of GC tissues and adjacent normal tissues was determined using quantitative Real Time-PCR (qRT-PCR) analyses. Survival plot was calculated using the Kaplan-Meier methods and log-rank test from the date of operation to the time of death or last follow-up date. The association between miR-1294 expression and clinical categorical data was analyzed using the chi-squared test. Moreover, Univariate and multivariate Cox analysis were performed to assess the risk factors of GC prognosis. RESULTS: We showed that miR-1294 expression was significantly downregulated in GC tissues compared to adjacent normal tissues. The low expression of miR-1294 in patients with GC was correlated with clinicopathological parameters including larger tumor size, lymph node metastasis, and distant metastasis. Kaplan-Meier survival analysis showed that GC patients with lower miR-1294 expression exhibited a shorter disease-free survival (DFS) and overall survival (OS) time compared to those patients with higher miR-1294 expression. Multivariate Cox analysis showed that lower miR-1294 expression, tumor size, lymph node metastasis, and distant metastasis were identified as independent risk factors of GC prognosis. CONCLUSIONS: Our results provided evidence that miR-1294 expression was significantly downregulated in GC and may serve as a predictor of GC prognosis.
