ABSTRACT
Objective: To clarify the mechanisms involvement in Alisertib-resistant colorectal cells and explore a potential target to overcome Alisertib-resistance. Methods: Drug-resistant colon cancer cell line (named as HCT-8-7T cells) was established and transplanted into immunodeficient mice. The metastasis in vivo were observed. Proliferation and migration of HCT-8-7T cells and their parental cells were assessed by colony formation and Transwell assay, respectively. Glycolytic capacity and glutamine metabolism of cells were analyzed by metabolism assays. The protein and mRNA levels of critical factors which are involved in mediating glycolysis and epithelial-mesenchymal transition (EMT) were examined by western blot and reverse transcription-quantitative real-time polymerase chain reaction(RT-qPCR), respectively. Results: In comparison with the mice transplanted with HCT-8 cells, which were survival with limited metastatic tumor cells in organs, aggressive metastases were observed in liver, lung, kidney and ovary of HCT-8-7T transplanted mice (P<0.05). The levels of ATP [(0.10±0.01) mmol/L], glycolysis [(81.77±8.21) mpH/min] and the capacity of glycolysis [(55.50±3.48) mpH/min] in HCT-8-7T cells were higher than those of HCT-8 cells [(0.04±0.01) mmol/L, (27.77±2.55) mpH/min and(14.00±1.19) mpH/min, respectively, P<0.05]. Meanwhile, the levels of p53 protein and mRNA in HCT-8-7T cells were potently decreased as compared to that in HCT-8 cells (P<0.05). However, the level of miRNA-125b (2.21±0.12) in HCT-8-7T cells was significantly elevated as compared to that in HCT-8 cells (1.00±0.00, P<0.001). In HCT-8-7T cells, forced-expression of p53 reduced the colon number (162.00±24.00) and the migration [(18.53±5.67)%] as compared with those in cells transfected with control vector [274.70±40.50 and (100.00±29.06)%, P<0.05, respectively]. Similarly, miR-125b mimic decreased the glycolysis [(25.28±9.51) mpH/min] in HCT-8-7T cells as compared with that [(54.38±12.70)mpH/min, P=0.003] in HCT-8-7T cells transfected with control. Meanwhile, in comparison with control transfected HCT-8-7T cells, miR-125b mimic also significantly led to an increase in the levels of p53 and β-catenin, in parallel with a decrease in the levels of PFK1 and HK1 in HCT-8-7T cells (P<0.05). Conclusions: Silencing of p53 by miR-125b could be one of the mechanisms that contributes to Alisertib resistance. Targeting miR-125b could be a strategy to overcome Alisertib resistance.
Subject(s)
Animals , Female , Mice , Humans , Azepines , Cell Line, Tumor , Cell Movement/genetics , Cell Proliferation , Gene Expression Regulation, Neoplastic , MicroRNAs/genetics , RNA, Messenger , Tumor Suppressor Protein p53/genetics , Drug Resistance, NeoplasmABSTRACT
OBJECTIVE@#The expression of microRNA-125b in tongue squamous cell carcinoma (TSCC) was detected and analyzed for its relationship with the clinicopathological features of TSCC.@*METHODS@#Real time fluorescence-quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of microRNA-125b in 35 TSCC tissues and adjacent normal tissues from 35 TSCC cases. The relationship between the expression of microRNA-125b in TSCC tissues and the clinicopathological features of patients with TSCC was analyzed. In situ hybridization (ISH) was used to detect the expression level of microRNA-125b gene in the TSCC tissues and adjacent normal tissues.@*RESULTS@#RT-qPCR results showed that the relative expression levels of microRNA-125b in the TSCC issues was 2.32±0.69, and that of normal tissues was 0.87±0.32. The statistical results showed that the expression level of microRNA-125b was significantly higher in the TSCC tissues than in the normal tissues (P<0.001). The expression level of microRNA-125b in the TSCC tissues was not significantly correlated with age, gender, pathological grade, and lymph node metastasis but was positively correlated with TNM stage. Patients with high TNM stage had high microRNA-125b expression levels (P<0.05). The ISH results showed that the expression levels of microRNA-125b in the TSCC tissues were 0.010±0.003, and that of normal tissues was 0.004±0.001. The expression levels of microRNA-125b in the 35 TSCC tissues were significantly higher than those in the normal tissues (P<0.05).@*CONCLUSIONS@#MicroRNA-125b is highly expressed in TSCC and associated with TNM stage, suggesting that high microRNA-125b expression may be involved in the development of TSCC.
Subject(s)
Humans , Carcinoma, Squamous Cell , Lymphatic Metastasis , MicroRNAs , Prognosis , Real-Time Polymerase Chain Reaction , Tongue NeoplasmsABSTRACT
Objective@#To observe the expression of microRNA-125b (miR-125), tumor necrosis factor alpha (TNF-α) and interleukin-6(IL-6) in premature rats exposed to hyperoxia.@*Methods@#Eighty 1-day old Sprague Dawley (SD) rats were randomly divided into an air group and a hyperoxia group.The rats in the hyperoxia group were continuously exposed to oxygen chamber for 1-3 L/min, oxygen volume fraction was maintained at (800±50) mL/L, and the rats in air group were placed in the same room with the oxygen volume fraction at 210 mL/L.The feeding conditions were same in 2 groups.Lung tissues of premature rats were extracted at different time (1, 4, 7, 10, 14 days). The pathologic changes in the lung tissues were observed by hematoxylin-eosin (HE) staining.The levels of miR-125b and TNF-α, IL-6 in lung tissues were detected by reverse transcription polymerase chain reaction (qRT-PCT) and enzyme-linked immunosorbent assay (ELISA).@*Results@#Compared with the air group, miR-125b in the hyperoxia group increased slowly after day 1, reached the highest in day 10 (2.554±0.323), and the relative expression in day 14 decreased slightly(2.329±0.263), and there were significant differences between 2 groups at di-fferent time (all P<0.05); in particular TNF-α level increased in day 7 [(78.55±39.53) ng/L], and reached the peak at day 10 [(80.16±11.24) ng/L], and there was a significant difference(P<0.05); IL-6 levels increased at day 7 [(45.44±31.94) ng/L], and reached the peak at day 10 [(90.38±8.24) ng/L], and there was a significant difference(P<0.05). There was a no significant correlation between miR-125b and TNF-α in the hyperoxia groups (r=0.132, P>0.05), but there was significant correlation between miR-125b and IL-6 in hyperoxia groups(r=0.439, P<0.05).@*Conclusions@#The levels of miR-125b, TNF-α and IL-6 are involved in the pathological process of bronchopulmonary dysplasia induced by hyperoxia, and IL-6 may be the key factor for miR-125b.
ABSTRACT
Objective To explore whether puerarin induced apoptosis of SKOV3 ovarian cancer cells by microRNA-125b.Methods Using the qRT-PCR technique to detect the change of microRNA-125b after puerarin pretreated SKOV3.Using RNA interference technology to inhibit microRNA-125b expression in SKOV3 cells.Using Western blot technique to detect apoptosis related proteins after microRNA-125b lower expression.Results Puerarin could significantly inhibit ovarian cancer cell SKOV3 proliferation activity and promote its apoptosis related proteins expression.And puerarin can promote the expression of microRNA-125b.Inhibition of microRNA-125b expression in ovarian cancer cell SKOV3 could reduce apoptosis protein expression in SKOV3 cell.Conclusion MicroRNA-125b was involved in puerarin induced SKOV3 cell apoptosis,and prompt microRNA-125b is key molecular of the drug resistance in SKOV3.
ABSTRACT
Objective To investigate the whether miR‐125b expression having the abnormality in nasopharyngeal carcinoma tissues and serum and whether having the correlation with the sensitivity of cisplatin chemotherapy .Methods The intact medical records and tissue and serum samples in 34 cases of clinically diagnosed nasopharyngeal carcinoma were collected and the serum samples in 34 individuals undergoing the physical examination were also collected as the controls .The expression of miRNA‐125b in carcinoma tissue ,paracancerous tissue and serum was detected by real‐time quantitative polymerase chain reaction (real‐time qPCR) .The statistical analysis was performed .Results The expression of miRNA‐125b in carcinoma tissue was significantly lower than that in paracancerous tissue(P=0 .006) ,and serum miRNA‐125b expression in nasopharyngeal carcinoma patients was signifi‐cantly lower than that in the individuals undergoing the healthy physical examination(P=0 .000);but the miRNA‐125b expression had no correlation between the paracancerous tissue and serum(r=0 .112 ,P=0 .528) .The expression of miRNA‐125b in cancer tis‐sues was correlated with the pathological typing ,T staging and N staging(P0 .05) .Conclusion The low miRNA‐125b expression may be involved in the occurrence and development of nasopharyngeal carcinoma and plays a certain significance for the diagnosis ,pathology typing and classification of nasopharyngeal carcinoma .Its expression level in cancer tissue can serve as one of indicators for screen‐ing out the cisplatin chemotherapy sche.