MicroRNA-150-5p inhibits the proliferation and invasion of human larynx epidermiod cancer cells though regulating peptidyl-prolyl cis/trans isomerase

Abstract Objective This study aimed to investigate the molecular mechanism of miR-150-5p regulating the malignant biological behavior of Human Epidermoid cancer cell (HEp-2) by targeting peptidyl-prolyl cis/trans isomerase NIMA-Interacting-1 (PIN1). Methods Firstly, qRT-PCR and Western blot were adopted to detect the expression levels of miR-150-5p and PIN1 in cancer tissue and paracancerous tissues of patients with LSCC, and those in human bronchial epithelial cells (16 HBE) and HEp-2. Next, the targeted relationship between miR-150-5p and PIN1 was assessed by bioinformatics website and dual-luciferase reporter assay, followed by their correlation analysis. Besides, after interfering with miR-150-5p or PIN1 expression in HEp-2 cells, CCK-8, cell colony formation assay, and transwell assay were utilized to detect the proliferation, viability, and invasion of cells, respectively. Subsequently, the protein levels of MMP-2, MMP-9, and EMT-related proteins in HEp-2 cells were checked by Western blot. Results Expression of miR-150-5p was down-regulated in LSCC tissues and HEp-2 cells. Moreover, miR-150-5p suppressed proliferation and invasion of HEp-2 cells, affected protein expression related to MMP and EMT, thereby inhibiting development of cancer. The expression of PIN1 was significantly increased in cancer tissues and HEp-2 cells, and there was a targeted relationship and negative correlation between miR-150-5p and PIN1 in cancer tissue. However, overexpression of PIN1 could reverse the effect of miR-150-5p on the proliferation and invasion of HEp-2 cells. Conclusion In a nutshell, there is a targeted relationship between PIN1 and miR-150-5p. Besides, miR-150-5p can inhibit the proliferation and invasion of HEp-2 cells by regulating the expression of PIN1. Level of evidence 3.

Effect of LINC00174 on the Malignant Proliferation of Multiple Myeloma Cells and Its Related Mechanism / 中国实验血液学杂志

OBJECTIVE@#To explore the biological function of LINC00174 in multiple myeloma (MM).@*METHODS@#Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expressions of LINC00174 and miR-150 in peripheral blood of MM patients and MM cell lines. EdU staining and flow cytometry were used to detect the effects of LINC00174 and miR-150 on the proliferation and apoptosis of MM cells. Western blot was used to detect the expressions of proliferation marker nuclear-related antigen Ki67, apoptosis-related protein cleaved caspase-3 and transcription factor forkhead box protein P1 (FOXP1). Bioinformatics and dual-luciferase reporter assay were used to verify the targeting relationship between LINC00174 and miR-150 and the targeting relationship between miR-150 and FOXP1.@*RESULTS@#The level of LINC00174 was significantly increased in peripheral blood of MM patients and MM cell lines (P <0.05). Compared with NC-siRNA group, the expression of LINC00174 was significantly reduced in LINC00174-siRNA group, the proliferation of U266 cells was reduced, the apoptosis rate was significantly increased, the level of Ki67 protein was reduced, and the level of cleaved caspase-3 protein was increased (all P <0.05). LINC00174 targeted regulation of the expression of miR-150. Compared with LINC00174-siRNA+NC inhibitor group, the expression of miR-150 in U266 cells in LINC00174-siRNA+miR-150 inhibitor group was significantly reduced, the cell proliferation was enhanced, the apoptosis rate was reduced, the level of Ki67 protein was increased, and the level of cleaved caspase-3 was decreased (all P <0.05). FOXP1 is the target gene of miR-150. Compared with NC mimic group, the expression of FOXP1 protein in miR-150 mimic group was significantly reduced, the cell proliferation was reduced, the apoptosis rate was significantly increased, Ki67 protein level was decreased, and the level of cleaved caspase-3 was increased. Compared with miR-150 mimic + vector group, the expression of FOXP1 protein in miR-150 mimic + pcDNA-FOXP1 group was significantly increased, the cell proliferation was enhanced, the apoptosis rate was reduced, the level of Ki67 protein was increased, and the level of cleaved caspase-3 was decreased (all P <0.05).@*CONCLUSION@#LINC00174 promotes the proliferation of MM cells and inhibits cell apoptosis by regulating the miR-150/ FOXP1 axis.

Effect of radiotherapy on the expression levels of miR-150-5p and miR-23a-3p in serum of tumor patients treated with tumor radiotherapy / 中华放射医学与防护杂志

Objective:To investigate the effect of irradiation on the expression of miR-150-5p and miR-23a-3p in human peripheral blood serum by collecting peripheral blood of tumor patients before and after radiotherapy, so as to provide scientific basis for finding radiation biomarkers.Methods:A total of 63 tumor patients treated with radiotherapy from October 2021 to March 2022 were enrolled in this study. The relative expression levels of miR-150-5p and miR-23a-3p in peripheral blood serum in these patients were detected using the real-time fluorescence quantitative PCR (qPCR) before and after radiotherapy. The differential changes in the expression levels of the two miRNAs in the peripheral blood serum of the patients before and after radiotherapy were compared, and their relationships with factors such as cancer types were analyzed.Results:The relative expression levels of miR-150-5p and miR-23a-3p in peripheral blood serum of the patients after radiotherapy were significantly lower than those before radiotherapy ( t = 4.97, Z = -2.77, P < 0.05). Among different cancer types, the relative expression level of miR-150-5p in the patients with breast cancer, esophageal cancer, or other digestive tract cancer decreased after radiotherapy ( t = 3.47, 2.47, 2.87, P < 0.05), and the relative expression level of miR-23a-3p in the patients with digestive tract cancer decreased after radiotherapy ( Z = -1.99, P < 0.05). The changes in the expression level of miR-150-5p before and after radiotherapy were not affected by gender, age, chemotherapy, and cancer type ( P > 0.05). By contrast, the changes in the expression level of miR-23a-3p before and after radiotherapy were significantly affected by gender, age, and chemotherapy ( t=2.04, -3.34, -2.29, P<0.05). Conclusions:The expression of miR-150-5p in the serum of tumor patients may be affected by radiotherapy, which has the potential to be used as a biological indicator of radiation.

Effects of urinary exosomal miR-150-5p on renal senescence in different age groups / 中华老年医学杂志

Objective:To investigate the expression of urinary exosomal miR-150-5p in different age groups and its relationship with kidney aging.Methods:Seventy-six subjects were recruited at the geriatrics department of Beijing Friendship Hospital from January 2014 to December 2016, with 15 in the young group, 35 in the middle-aged group, and 26 in the elderly group.We detected the expression of urinary exosomal miR-150-5p by real-time PCR from urine samples of different age groups and used Logistic regression analysis to evaluate the correlation of different levels of miR-150-5p with age, gender, history of hypertension, history of coronary heart disease, and hyperlipidemia.Additionally after overexpression and knockdown of miR-150-5p in human renal tubular cells of the HK2 cell line, we analyzed the expression levels of epithelial-to-mesenchymal transition(EMT)-related proteins by Western blotting.Results:Logistic regression analysis showed that age was closely related to estimated urinary exosomal miR-150-5p levels( P<0.05), and estimated urinary exosomal miR-150-5p levels in the elderly group(1.33±0.41)were significantly higher than those in the young group(0.88±0.40)( P<0.05). In addition, estimated urinary exosomal miR-150-5p levels became inversely proportional to the glomerular filtration rate as age increased. In vitro experiments showed that the expression levels of ZO-1 and E-cadherin in epithelial cells were slightly decreased after knockdown of miR-150-5p in renal tubular epithelial cells of the HK-2 cell line. Conclusions:The expression of miR-150-5p in urinary exosomes may be slightly increased in the elderly.It could inhibit the occurrence of renal EMT, and its expression is correlated with renal function.Mir-150-5p may also affect the expression of adhesion molecule-related proteins in HK-2 cells.MiR-150-5p in urinary exosomes is expected to become one of the genetic markers associated with renal aging.

MiR-150-5p protects rats from middle cerebral artery occlusion by inhibiting the toll-like receptor-5/nuclear factor-kappa B p65 signaling pathway / 中国组织工程研究

BACKGROUND: There is an inflammatory response in the lesion tissue of ischemic cerebral infarction, and the expression of miR-150-5p is significantly decreased. Whether miR-150-5p inhibits the release of inflammatory factors and alleviates the injury of ischemic cerebral infarction tissue through the Toll-like receptor-5/nuclear factor-KB pathway remains unclear. OBJECTIVE: To investigate the role and preliminary mechanism of miR-150-5p in ischemic cerebral infarction in rats. METHODS: (1) The rat models of middle cerebral artery occlusion were constructed and the rat models were divided into five groups: Control, miR-150-5p agomir, agomir control, miR-150-5p antagomir and antagomir control groups. (2) The rats in the control group was given the intracerebroventricular injection of normal saline, and the rats in the latter four groups were given the intracerebroventricular injection of miR-150-5p agomir (miR-150-5p agonist), agomir negative control, miR150-5p antagomir (miR150-5p inhibitor) and antagomir negative control, respectively. (3) After 7 days, the brain was graded by modified neurological severity score, the cerebral infarct volume was measured by MRI, and the histopathological changes were observed by hematoxylin-eosin staining. The expression levels of miR-150-5p, interleukin-6, tumor necrosis factor-a, Toll-like receptor-5 and nuclear factor-KB p65 in brain tissues were detected by qRT-PCR, ELISA and western blot assay, respectively. The target relationship between miR150-5p and Toll-like receptor-5 was verified by luciferase assay by retrieving the bioinformatics website Targetscan to predict the binding sites of miR-150-5p and Toll-like receptor-5. RESULTS AND CONCLUSION: (1) Compared with the control group, the modified neurological severity score, and levels of interleukin-6, tumor necrosis factor-a, Toll-like receptor-5 and nuclear factor-KB p65 proteins were significantly decreased in the miR-150-5p agomir group (P 0.05). (3) TargerScan website prediction results and luciferase reporter gene analysis results showed that miR-150-5p and Toll-like receptor-5 had a targeted binding site. (4) These results imply that miR-150-5p can inhibit the inflammatory signaling pathway of Toll-like receptor-5/nuclear factor-KB p65 in brain injury caused by ischemia and reduce the inflammatory response, thereby alleviating the damage of nerve function and playing a protective role.

Effect of microRNA-150 on proliferation,apoptosis,invasion and metastasis of epithelial ovarian cancer cells / 实用肿瘤学杂志

Objective The aim of this study was to investigate the expression of microRNA-150(miR-150)in human epi-thelial ovarian cancer cells and its effect on proliferation,apoptosis,invasion and metastasis of human epithelial ovarian cancer cells. Methods The expression level of miR-150 in cells from each treatment group was detected by Real-Time PCR(qRT-PCR);effects of proliferation,apoptosis,invasion and metastasis of epithelial ovarian cancer cells was investigated by MTT,flow cytometry, and transwell assays. Results Compared with normal ovarian epithelial cells(T29),the expression of miR-150 was significantly de-creased in epithelial ovarian cancer cells(A2780 and OVCAR3)(P<0. 01); After transfection miR-150 mimic,the expression of miR-150 in A2780 and OVCAR3 cells was significantly increased(P<0. 01);After 3 d of transfection,the OD values of the miR-150mimicgroup(A2780:1.12±0.03;OVCAR3:1.91±0.03)werelowerthanthatintheblankgroup(A2780:2.35±0.09;OVCAR3:2.63 ±0.07)and the miR-150 NC group(A2780:2.18 ±0.07;OVCAR3:2.43 ±0.11)(P<0.01);The apoptotic rate in the miR-150 mimic group(A2780:16. 10 ± 0. 58% ;OVCAR3:15. 16 ± 1. 30% ) were significantly increased when compared to the blank group(A2780:10. 07 ± 0. 66%;OVCAR3:3. 81 ± 0. 24%) and the miR -150 NC group(A2780:10. 36 ± 1. 08%;OVCAR3:4.89 ±0.07%)(P<0.01);The number of transmembrane cells in the miR-150 mimic group(A2780:38.67 ±2.03;OVCAR3:28. 67 ± 2. 03)was higher than that in the blank group(A2780:76. 30 ± 7. 45;OVCAR3:55. 67 ± 3. 18)and the miR-150 NC group(A2780:74. 33 ± 5. 78;OVCAR3:56. 33 ± 3. 84)(P<0. 01). Conclusion The decreased expression of miR-150 in epi-thelial cancer cells may be one of the mechanisms of proliferation,invasion and metastasis of epithelial ovarian cancer. Up-regulation of miR-150 may inhibit the proliferation of epithelial ovarian cancer cells and promote apoptosis to reduce the abilities of invasion and metastasis in epithelial ovarian cancer cells.

Expression and Clinical Significance of Plasma miR-150,SCCA and CA125 in Patients with HPV Positive Cervical Cancer and CIN / 现代检验医学杂志

Objective To investigate the expression and clinical significance of plasma miR-150,squamous cell carcinoma anti gen (SCCA) and carbohydrate antigen 125 (CA125) in human papillomavirus (HPV) positive cervical cancer patients.Methods RT PCR was detected in 108 cases of HPV patients with positive cervical cancer (cervical cancer group) and 40 cases of HPV positive CIN patients (CIN group) and 40 cases of HPV with positive/HPV positive uterine benign lesions (control group),plasma miR-150,SCCA and CA125 levels.Analyzed the relation between miR 150 expression and clinicopathological features of cervical cancer.ROC curve was used to evaluate the early diagnostic value of miR-150,SCCA and CA125 in patients with HPV positive cervical cancer.Multivariate Logistic regression model was used to analyze the rela tionship between three indicators and HPV positive cervical cancer.Results The levels of miR-150,SCCA and CA125 in the cervical cancer group were significantly higher than those in the CIN group and the control group [miR-150(2-△△Ct):10.28 ±4.16 vs 4.72± 1.18 and 1.83±0.64.SCCA(ng/ml):9.86±2.14 vs 3.18±0.85 and 1.35±0.3t.CA125 (U/ml):46.26 ±11.58 vs 19.14±7.05 and 16.42±5.83,F=16.427,13.205,8.719,all P＜0.01].The expression of plasma miR 150 in HPV positive cervical cancer correlated with clinical stage,lymph node metastasis and depth of invasion (P＜0.05).The optimal cut-off values of plasma miR-150 for diagnosis of HPV positive cervical cancer and CIN were 8.25 and 3.46,and the sensitivity and specificity were 87.0 ％ and 84.3 ％,81.0 ％ and 80.2 ％,respectively.Logistic regression analysis showed that elevated plasma miR 150 and SCCA levels were independent risk factors for HPV positive cervical cancer [OR(95 ％ CI)=2.107(1.915～2.814),OR(95％CI)=1.583(1.327～2.036)].Conclusion Plasma miR 150 was significantly up regulated in patients with HPV positive cervical cancer,which can be used as a biological marker for the early diagnosis of HPV positive cervical cancer and the differential diagnosis of CIN.

Regulatory effect of hsa-miR-150-5p on malignant behaviors of glioblastoma U-251MG cells by targeting HIF1α / 中国肿瘤生物治疗杂志

@#Objective: To explore the effect and possible mechanisms of has-miR-150-5p targeting HIF1α to regulate malignant biological behaviors of glioblastoma (GBM) U-251MG cells. Methods: Real-time quantitative PCR (RT-PCR) was used to detect the expression of miR-150-5p and hypoxia inducible factor 1 (HIF1α) in U-251MG cells. Luciferase report assay was carried out to verify the biological relationship between miR-150-5p and HIF1α and their biological functions in U-251MG cells. The protein expressions of miR150-5pand HIF1α in U-251MG cells were detected by western blotting. The ability of cell migration was detected by wound healing test and cell invasion ability was detected by transwell test. Results: After miR-150-5p mimic transfection, the mRNA expression of HIF1α was significantly reduced in U-251MG cells (P<0.01). Bioinformatics prediction and luciferase reporter assay demonstrated that miR-150-5p down-regulated HIF1α through directly binding to HIF1α 3’-untranslated region (3’-UTR) (all P<0.05). In U-251MG cells, miR-150-5p over-expression significantly inhibited HIF1α expression, cell invasion and migration (all P<0.05). Conclusion: miR150-5p inhibits cell invasion and metastasis through negative regulation of HIF1α, indicating that miR-150-5p and HIF1α were both potential therapeutic targets for glioblastoma.

Study on the Levels and Clinical Significance of Serum miR-150 in Children with Nephrotic Syndrome / 现代检验医学杂志

Objective To investigate the levels and clinical significance of serum miR-150 in children with primary nephrotic syndrome(NS).Methods Serum samples were collected from 78 NS children and 79 age-and sex-matched control children in Nanjing General Hospital,Jiangsu Province Hospital of TCM and Nanjing Children Hospital from March 2010 to May 2014.Quantitive Real-time PCR(qRT-PCR)assays were used to determine the concentrations of serum miR-150 in NS and control children.The other lipid and renal function parameters including serum TP,ALB,GLO,TC,TG,Urea,Cr,Uric acid and Uric protein were also assessed.Statistical analyzes were used to evaluate the clinical value of serum miR-150 for NS as well as to assess the clinical association between the levels of serum miR-150 and other clinical parameters.Results The ser-um levels of miR-150 were significantly elevated in NS children[101.4(21.29~336.6)fmol/L(F=3.658,P<0.001)]as compared with controls[34.11(5.53~134.2)fmol/L].ROC curve analysis showed that the area under the receiver operat-ing characteristic curve(AUCROC)was 0.892(95% CI=0.843~0.940).Spearman rank correlations analyzes showed that the levels of serum miR-150 were significantly negatively associated with GLO(r=-0.231,P=0.042)and TG(r=-0.233,P=0.040)in NS children.Multiple linear regression analyses showed that serum miR-150 was independently associ-ated with serum ALB levels(β=0.241,P=0.034;adjusted r2=0.046)after adjustment of other related factors.Further-more,multivariate logistic regression analysis showed that serum miR-150 an independent risk factor for NS after adjusting other factors including age and gender[OR=16.07(95% CI=5.35~48.28),P<0.001].Conclusion The serum levels of miR-150 were markedly elevated in NS children and closely associated with with impaired kidney function as well as lipid pa-rameters,and may have the potential as a novel auxiliary diagnosis marker for assessing the development of NS.

miR-150 deletion affects breeding and hematologic parameters of mice / 中国免疫学杂志

Objective:To investigate the effect of miR-150 gene deletion on the breeding and hematologic parameters of mice. Methods:The nest litter size,wean rate and weight changes of miR-150 knock out (miR-150ko) and C57BL/6J mice were com-pared. The hematology indexes were analyzed by automated blood cell counter, the serum biochemical parameters were analyzed by automatic biochemical analyzer. Results:The nest litter size and wean rate of miR-150ko mice were significantly decreased compared with that of C57BL/6J mice. The number of total white blood cells,intermediate cells,neutrophils,and the percentage of neutrophils and intermediate cells were significantly increased in miR-150ko mice compared with that of C57BL/6J mice. However,the number and per-centage of platelets and lymphocytes decreased significantly in miR-150ko mice. In addition,the levels of serum glucose and TC were in-creased significantly in miR-150ko mice compared with that of C57BL/6J mice. Conclusion: miR-150 gene deletion impairs the breeding and has complex impact on hematologic parameters of mice.

MiR-150 downregulation and its significance in mantle cell lymphoma / 中国肿瘤临床

Objective: This study explores the miR-150 expression and its clinical significance in mantle cell lymphoma (MCL). Methods: The miR-150 and c-Myc expression was measured in 29 primary MCL tissue samples and 7 normal donors through quantita-tive real-time polymerase chain reaction. MiR-150 expression was detected in Mino and HBL-2 cells after c-Myc was knocked down by small interfering RNAs. MiR-150 was analyzed in tet-treated P493-6 cells with Myc turned off. The number of tumor colonies in the BL-2 cells transfected with pre-miR-150 was determined through colony formation assay, and c-Myb was detected through Western blot. Results: Compared with the normal donors, miR-150 expression was significantly downregulated and c-Myc was considerably overexpressed in MCL. Moreover, MCLs with high Myc expression had a significantly low miR-29 expression. MiR-150 was upregu-lated in the Mino and HBL-2 cells with c-Myc knockdown. MiR-150 was evidently upregulated in P493-6 cells after c-Myc was turned off. MiR-150 overexpression suppressed colony formation and c-Myb expression of HBL-2. Conclusion: MiR-150 is downregulated in MCL, which may be related to c-Myc overexpression. The recovery of miR-150 suppresses MCL cell survival. These results indicate that miR-150 may be a potential therapeutic target of MCL.