miR-155 Research Articles

The role of miR-145 in promoting the fibrosis of pulmonary fibroblasts.

The effects of miR-145 (microRNA 145) on M. pneumoniae (MP)-infected MRC-5 (Medical Research Council cell strain 5) cell TGF-β/Smad (transforming growth factor beta/Smad) fibrosis pathway were explored through constructing MP-infected MRC-5 cell models. In addition, the qPCR (quantitative real-time polymerase chain reaction) and Western blot were applied to detect the mRNA and protein expressions of miR-145, TGF-β1 (transforming growth factor beta 1), Smad3, Smad4, MMP2 (matrix metalloproteinase 2), FN1 (fibronectin 1), ELN (elastin) and COLI α1 (collagen type I alpha 1) signaling molecules in TGF-β/Smad fibrosis pathway. The results showed that the expression of miR-145 in MRC-5 cells was significantly increased after MP infection. In addition, miR-145 inhibited the fibrosis promoting TGF-β/Smad pathway by targeting Smad3, a key factor in the TGF-β/Smad pathway. It can be concluded that, in the process of MP infection, the expression of miR-145 is stimulated to negatively regulate the fibrosis-promoting pathway of TGF-β/Smad.

Association of genetic polymorphisms of miR-145 gene with telomere length in omethoate-exposed workers

Omethoate, an organophosphorous pesticide, causes a variety of health effects, especially the damage of chromosome DNA. The aim of the study was to assess the correlation between polymorphisms of encoding miRNA genes and telomere length in omethoate–exposure workers. 180 workers with more than 8 years omethoate–exposure and 115 healthy controls were recruited in the study. Genotyping for the selected single nucleotide polymorphisms loci were performed using the flight mass spectrometry. Real-time fluorescent quantitative polymerase chain reaction(PCR) method was applied to determine the relative telomere length(RTL) in human peripheral blood leukocytes DNA. After adjusting the covariate of affecting RTL, covariance analysis showed that the female was significantly longer than that of the male in control group(P < 0.046). For the miR-145 rs353291 locus, this study showed that RTL of mutation homozygous AG+GG individuals was longer than that of wild homozygous AA in the exposure group (P = 0.039). In the control group, RTL with wild homozygous TT genotype in miR-30a rs2222722 polymorphism locus was longer than that of the mutation homozygous CC genotype (P = 0.038). After multiple linear regression analysis, the independent variables of entering into the model were omethoate–exposure (b = 0.562, P < 0.001), miR-145 rs353291 (AG+GG) (b = 0.205, P = 0.010). The prolongation of relative telomere length in omethoate exposed workers was associated with AG+GG genotypes in rs353291 polymorphism of encoding miR-145 gene.

Differential expression of exosomal miRNAs between androgen-dependent prostate cancer cell and androgen-independent prostate cancer cell

Objective Studying the differential expression of exosomal miRNAs between androgen-dependent prostate cancer cell and androgen-independent prostate cancer cell, in order to further elucidate the mechanism of androgen-independent prostate cancer and find new targets for its treatment. Methods Prostate cancer androgen-dependent LNCaP cell and prostate cancer androgen-independent LNCaP-AI+F cell (induced by androgen and flutamide) were selected as the study subjects. Illumina HiSeqTM 2500 was used to perform high-throughput sequencing between the two groups. The differentially expressed exosomal miRNAs was verified by quantitative real-time PCR(qRT-PCR). The difference was statistically significant by t-test. Results Through the analysis of high-throughput sequencing results, thirteen molecules were screened increased in extracellular exosomes of the androgen-independent cell, including miR-7-5p, let-7a-5p, miR-375, miR-423-3p, miR-378a-3p, and miR-92a-3p, etc. Among them, miR-7-5p was verified by quantitative real-time PCR to be up-regulated by 19.52-fold (t=9.857, P=0.001). Conclusion Differentially expressed exosomal miRNAs may predict the development of androgen-independent prostate cancer and may further regulate the development of androgen-independent prostate cancer. Key words: Prostatic neoplasms; Exosomes; microRNAs; High-throughput nucleotide sequencing

Effect of microRNA-221 targeting HMBOX1 on biological function of gastric cancer cells

Objective To explore the target of microRNA (miRNA, miR)-221p and its effect on biological function of gastric cancer cells. Methods Bioinformatics was used to predict the possible targets of miR-221, and plenti-miR-221 overexpression vector was constructed. Lentiviruses were packaged. MiR-221 stable expression cell line (observation group) and control cell line (control group) was established by lentiviruses infecting. The expression of miR-221 target gene protein in control and observation groups were analyzed by Western blotting. The proliferation ability in control and observation groups was detected by cell counting kit-8 (CCK-8). The invasive ability was analyzed by Transwell in control and observation groups. The apoptosis level of control and observation groups was analyzed by flow cytometry. The tumor growth and metastasis of the control group and the observation group were analyzed. Results Bioinformatics analysis showed that the target gene of miR-221 was HMBOX1. Compared with the control group (1.12±0.23), the expression of HMBOX1 protein in the observation group (0.32±0.12) was significantly decreased (t=3.976, P<0.01). Compared with the control group, the proliferation ability of cells in the observation group increased significantly (F=3.102, P<0.01). Transwell quantitative analysis showed that the invasive ability of cells in the observation group [(125.43±15.32) cells] was significantly higher than that in the control group [(57.40±9.81) cells], and the difference was statistically significant (t=3.190, P<0.01). The level of apoptosis in the observation group [(12.78±3.56)%] was significantly lower than that in the control group [(25.43±7.54)%, t=2.918, P<0.01]. The growth rate of cells in the observation group was significantly higher than that in the control group (F=3.012, P<0.01). The number of intraperitoneal metastatic lesions in the observation group (15.26±3.29) was significantly higher than that in the control group (6.19±2.01) (t=2.991, P<0.01). Conclusion MiR-221 targeting HMBOX1, promote the proliferation and invasion of tumor cells, reduce cell apoptosis, and promote the occurrence and metastasis of tumor. Key words: MicroRNA-221; HMBOX1; Proliferation; Apoptosis; Metastasis

Acupuncture preconditioning protects against myocardial ischemia/reperfusion injury mediated apoptosis through miR-214/NCX1 activation: a hypothesis

Acupuncture preconditioning protects against myocardial ischemia/reperfusion injury mediated apoptosis through miR-214/NCX1 activation: a hypothesis

MiR-205-3p、miR-205-5p对头颈鳞状细胞癌细胞增殖、迁移及侵袭的影响

Objective To investigate the effects of microRNA-205-3p (miR-205-3p) and microRNA-205-5p (miR-205-5p) on the cell proliferation, migration and invasion of head and neck squamous cell carcinoma (HNSCC). Methods The miRNA expression data were obtained from The Cancer Genome Atlas, and were used to determine miR-205 expression in HNSCC and paracancerous tissues. Quantitative real-time fluorescent polymerase chain reaction (qRT-PCR) was used to detect the expressions of miR-205-3p and miR-205-5p in HNSCC cell lines 5-8F, 6-10B, CNE2, Tu686 and mucosal tissues of nasopharynx. MiR-205-3p inhibitor, miR-205-5p inhibitor and miR-NC were transfected into Tu686 cells, and qRT-PCR was employed to evaluate the silence efficiency. In the following study, the cells were divided into four groups: miR-205-3p inhibitor and control, miR-205-5p inhibitor and control. Then the cell counting kit (CCK-8) assay, Transwell migration and invasion assays were carried out to examine the proliferation, migration and invasion abilities of the cells in the four groups. Results Compared with paracancerous tissues, the higher expression of miR-205 in HNSCC tissues was observed [M (QR): 61 012 (51 448) vs. 28 579 (35 959), Z=-6.420, P<0.001)]. The expression levels of miR-205-3p in HNSCC cell lines 5-8F, 6-10B, CNE2, Tu686 and mucosal tissues of nasopharynx were 0.36±0.07, 0.20±0.06, 0.15±0.04, 0.25±0.04 and 1.00±0.00 respectively, with a significant difference (F=162.71, P<0.001). The expression levels of miR-205-5p in cell lines 5-8F, 6-10B, CNE2, Tu686 and mucosal tissues of nasopharynx were 0.20±0.01, 0.21±0.01, 1.06±0.18, 23.61±2.07 and 1.00±0.00 respectively, with a significant difference (F=371.81, P<0.001). Compared with 5-8F, 6-10B, CNE2 cells, the expression of miR-205-3p in Tu686 cells was higher than those in 6-10B, CNE2 cells (P=0.195; P=0.020), and lower than that in 5-8F cells (P=0.023), and the expression of miR-205-5p in Tu686 cells was the highest (P<0.001; P<0.001; P<0.001). The expressions of miR-205-3p and miR-205-5p in Tu686 cells were effectively downregulated by inhibitors, and the silence efficiencies were 87% and 83%, respectively. The absorbance (A) values of miR-205-3p inhibitor group on the first, second, third, fourth and fifth day were 0.26±0.06, 0.55±0.11, 1.52±0.13, 1.91±0.07, 2.14±0.24, and those of miR-NC group were 0.29±0.07, 0.78±0.11, 1.59±0.15, 1.95±0.08, 2.02±0.12. There were no significant differences between the two groups (t=0.506, P=0.639; t=2.459, P=0.070; t=0.573, P=0.597; t=0.655, P=0.548; t=-0.759, P=0.490). The cell proliferations of miR-205-5p inhibitor group on the first, second, third, fourth and fifth day were 0.30±0.08, 0.61±0.08, 0.85±0.08, 1.08±0.12, 1.16±0.18, and those of miR-NC group were 0.41±0.10, 0.78±0.14, 1.33±0.28, 1.87±0.09, 2.08±0.19. The cell proliferations of miR-205-5p inhibitor group on the third, fourth and fifth day were significantly lower than those of miR-NC group (t=3.665, P=0.017; t=12.223, P<0.001; t=7.825, P<0.001). Transwell migration assay demonstrated that downregulation of miR-205-3p had no significant effect on migration abilities of Tu686 cells. The numbers of cell permeating septum of miR-205-3p inhibitor group and miR-NC group were 192.00±28.49 and 188.40±22.52, respectively. There was no significant difference between the two groups (t=-0.160, P=0.877). Downregulation of miR-205-5p significantly decreased the migration abilities of Tu686 cells. The numbers of cell permeating septum of miR-205-5p inhibitor group and miR-NC group were 109.40±27.63 and 183.60±31.63, respectively, and the difference was statistically significant (t=3.951, P=0.004). Transwell invasion assay showed that the numbers of cell permeating septum of miR-205-3p inhibitor group and miR-NC group were 93.40±10.24 and 96.20±16.56, respectively. There was no significant difference between the two groups (t=0.322, P=0.756). The numbers of cell permeating septum of miR-205-5p inhibitor group and miR-NC group were 53.00±17.80 and 94.40±14.38, respectively, and the difference was statistically significant (t=4.045, P=0.004). Conclusion Downregulation of miR-205-5p can inhibit the proliferation, migration and invasion abilities of Tu686 cells. However, downregulation of miR-205-3p has no significant effect on the proliferation, migration and invasion of Tu686 cells. Key words: Head and neck neoplasms; MicroRNAs; Cell proliferation; Cell movement; Neoplasm invasiveness

Research progress of miR-224 in colorectal cancer

Colorectal cancer is the third most common cancer in humans and poses a serious threat to human health. MicroRNAs (miRNAs) affected the mechanism of occurrence and development of colorectal cancer.As an important member of miRNA, miR-224 modulates the migration, proliferation, metastases and drug resistance of colorectal cancer by acting on its downstream targets. The expression of miR-224 affects the prognosis of colorectal cancer. Therefore, this review summarizes the role of miR-224 and its downstream targets in colorectal cancer, it may provide a new method for diagnosis and treatment to colorectal cancer at the cellular and molecular level. Key words: Target gene repair; Colorectal neoplasms; Review; MiR-224

Research progress of microRNA-155 and microRNA-181b in chronic lymphocytic leukemia

Chronic lymphocytic leukemia (CLL) is characterized by the aggregation of function defect of B cells, and the onset of CLL is occult. CLL can be divided into inertness CLL and invasion CLL according to the course of disease, and their prognosis exist great differences. MicroRNAs (MiRNA) are closely linked with B cells development and play roles in regulation B cells development by targeting the corresponding transcription factors. Recent studies show that MiRNA-155 and MiRNA-181b play important roles in the process of CLL generation and development, especially with poor prognosis of CLL. This paper reviews the research progresses of biological characteristics of MiRNA-155 and MiRNA-181b in CLL diagnosis and treatment. Key words: Leukemia, lymphocytic, chronic, B-Cell; MicroRNAs; MiR-155; MiR-181b; Prognosis

Molecular mechanism of microRNA-145 targeting protein kinase Bγ in regulating growth and metastasis of thyroid cancer cells

Objective To investigate the expression of microRNA (miRNA)- 145 and protein kinase Bγ (Akt3) in thyroid carcinoma and the molecular mechanism of miRNA-145 targeting Akt3 inhibiting the proliferation and invasion of thyroid carcinoma cells. Methods Real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) was used to detect the expression of microRNA-145 and Akt3 in 49 fresh thyroid carcinoma tissues and adjacent tissues. The relationship between the expression of microRNA-145 and Akt3 was analyzed. TPC-1 thyroid cancer cell line was used as the research object. The effects of microwave irradiation on the growth, invasion and metastasis of thyroid cancer cells were detected by methyl thiazol tetrazolium (MTT) and Transwell invasion assays. The expression levels of microwave irradiation-145 and Akt3 were detected by Western blotting. Results The results of FQ-PCR showed that the relative expression levels of miRNA-145 in thyroid carcinoma and adjacent tissues were 1.65±0.33 and 3.13±0.53, and those of Akt3 were 2.96±0.85 and 0.83±0.32, respectively. The difference was statistically significant (P=0.000). The relative expression levels of microRNAs-145 in TPC-1 cells and primary thyroid follicular cells were 1.91±0.57 and 3.47±0.59, and those of Akt3 were 3.62±1.04 and 0.76±0.32, respectively. There was statistically significant difference (P=0.000). The increase of A570 in TPC-1 cells transfected with anti-miR-145 was the fastest, and that in TPC-1 cells transfected with analogues of miR-145 was the slowest (P=0.000). As compared with TPC-1 cells without transfection and TPC-1 cells transfected only with liposome 2000, the number of invasive cells transfected with anti-miR-145 increased significantly (P=0.000), while the number of invasive cells transfected with micR-145 analogues decreased significantly (P=0.000). The expression levels of p-Akt and Akt-3 protein in miR-145-mimics-transfected and non-transfected TPC-1 cells were significantly lower than those in TPC-1 cells transfected with anti-miR-145. Conclusion MiR-145 can target down-regulate the expression of Akt3 to inhibit the proliferation and invasion of thyroid cancer cells. Key words: Thyroid cancer; MicroRNA-145; Protein kinase Bγ; Proliferation; Metastasis

Inhibition of MicroRNA-221 and 222 Enhances Hematopoietic Differentiation from Human Pluripotent Stem Cells via c-KIT Upregulation.

The stem cell factor (SCF)/c-KIT axis plays an important role in the hematopoietic differentiation of human pluripotent stem cells (hPSCs), but its regulatory mechanisms involving microRNAs (miRs) are not fully elucidated. Here, we demonstrated that supplementation with SCF increases the hematopoietic differentiation of hPSCs via the interaction with its receptor tyrosine kinase c-KIT, which is modulated by miR-221 and miR-222. c-KIT is comparably expressed in undifferentiated human embryonic and induced pluripotent stem cells. The inhibition of SCF signaling via treatment with a c-KIT antagonist (imatinib) during hPSC-derived hematopoiesis resulted in reductions in the yield and multi-lineage potential of hematopoietic progenitors. We found that the transcript levels of miR-221 and miR-222 targeting c-KIT were significantly lower in the pluripotent state than they were in terminally differentiated somatic cells. Furthermore, suppression of miR-221 and miR-222 in undifferentiated hPSC cultures induced more hematopoiesis by increasing c-KIT expression. Collectively, our data implied that the modulation of c-KIT by miRs may provide further potential strategies to expedite the generation of functional blood cells for therapeutic approaches and the study of the cellular machinery related to hematologic malignant diseases such as leukemia.

Expression and significance of miR-126 and VEGF in proliferative diabetic retinopathy

Objective To investigate the expression and clinical significance of miR-126 and vascular endothelial growth factor (VEGF) in proliferative diabetic retinopathy (PDR). Methods 226 cases of diabetic retinopathy (DR) patients admitted in our hospital were studied, including 110 cases of PDR (group PDR), 116 non proliferative diabetic retinopathy (NPDR) (group NPDR). 80 patients with diabetes mellitus without retinopathy (NDR) were enrolled in DR group at the same period, another 80 healthy subjects (control group) were selected as control group. The plasma miR-126 level of all subjects was detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). The enzyme linked im-munosorbent assay (ELISA) method was used to detect plasma VEGF level. The clinical diagnostic value of miR-126 and VEGF in PDR patients was further analyzed. Results Plasma levels of miR-126 in PDR group, NPDR group and NDR group were lower than those in control group (P<0.05), PDR group was lower than NPDR group and NDR group (P<0.05); plasma levels of VEGF in PDR group, NPDR group and NDR group were higher than those in control group (P<0.05), PDR group was higher than NPDR group and NDR group (P<0.05). Total cholesterol (TC), triglyceride (TG) and glycated hemoglobin (HbA1c) in PDR group were higher than those in NPDR group and NDR group (P<0.05), low density lipoprotein cholesterol (LDL-C) in PDR group, NPDR group and NDR group were higher than those in control group (P<0.05), and LDL-C in PDR group was higher than that in NPDR group and NDR group (P<0.05). High density lipoprotein cholesterol (HDL-C) in PDR group and NDR group was lower than that in control group (P<0.05), C-reactive protein (CRP) in PDR group, NPDR group and NDR group was higher than that in control group (P<0.05); plasma miR-126 levels in PDR group were negatively correlated with TC, LDL-C, CRP and HbA1c (P<0.05), but positively correlated with HDL-C (P<0.05), and no correlation with TG; the plasma levels of VEGF in PDR patients were positively correlated with TC, TG, LDL-C, CRP and HbA1c (P<0.05), but negatively correlated with the expressions of miR-126 (r=-0.573, P=0.000); the AUC of miR-126 was 0.861, and when the cut-off value was <0.64, the diagnostic sensitivity and specificity were 82.50% , 83.64%; the area under curve (AUC) of VEGF was 0.889, and when the cut-off value was <7.000, the sensitivity and specificity of diagnosis were 82.73%, 86.25%; 0.847 for the AUC of HbA1c, and the sensitivity and specificity 81.82%, 87.50% respectively. Conclusions Plasma miR-126 is low-expressed and VEGF is high-expressed in PDR patients, there is a negative correlation between the two indexes. They may be involved in the course of PDR through abnormal lipid metabolism and inflammatory reaction and may be a potential biomarker for early diagnosis of PDR. Key words: Diabetic retinopathy/BL; MicroRNAs/BL; Vascular endothelial growth factors/BL

Aberrant expression of circular RNA in mesenchymal stem cells from psoriatic skin lesions

Objective To investigate the role of circular RNA (circRNA) in the occurrence of psoriasis. Methods Mesenchymal stem cells were isolated from skin lesions of 15 patients with psoriasis (psoriasis group) and skin tissues of 15 healthy human controls (control group) separately, and then subjected to cultivation. Flow cytometry and cell differentiation assay were performed to identify these mesenchymal stem cells. RNA sequencing was conducted to measure the expression of circRNA, and detailed bioinformatics analysis was performed. Then, 7 differentially expressed circRNAs were selected, and a circRNA-microRNA interaction network was established. In this network, 3 related microRNAs were selected and validated by real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) . Statistical analysis was carried out by a two-sample t-test for comparison of qRT-PCR results between the psoriasis group and control group. Results As RNA sequencing showed, a total of 6 323 circRNAs were identified, and 3 227 out of these circRNAs were first reported in this study. Compared with the control group, 129 circRNAs were differentially expressed in the psoriasis group, including 123 up-regulated circRNAs and 6 down-regulated circRNAs. The predicted circRNA-microRNA interaction network showed that several psoriasis-related microRNAs were associated with the 7 circRNAs, such as miR-17-5p, miR-30e-5p, miR-142-3p/5p, miR-369-3p, miR-184, miR-4490, miR-654-3p and miR-423-5p. qRT-PCR also confirmed that the 7 differentially expressed circRNAs were up-regulated in the psoriasis group. Compared with the control group, the expression of the 3 selected microRNAs significantly decreased in the psoriasis group (t=3.993, 3.217, 2.918, respectively, all P < 0.05) . Conclusion There is aberrant expression of circular RNAs in the mesenchymal stem cells from psoriatic skin lesions, which may take part in the occurrence of psoriasis. Key words: Psoriasis; Mesenchymal stem cells; Circular RNA

Relationship between drug sensitivity of gastric cancer tissues to L-oxaliplatin and microRNA-150

Objective To explore relationship between drug sensitivity of gastric caner tissues to L-oxaliplatin (L-OHP) and microRNA (miRNA, miR)-150 and its significance. Methods Specimens of gastric cancer tissues and cancer-adjacent normal tissues from 80 patients were collected. The expression levels of miR-150 and mRNA of multidrug resistance gene 1 (MDR1), glutathione S transferase-π (GST-π), B cell lymphoma/leukemia-2 (bcl-2) and Survivin were detected by real-time quantitative polymerase chain reaction (Real-time PCR), and those of P-glycoprotein (P-gp), GST-π, bcl-2 and Survivin proteins were detected by immunohistochemistry. Methyl thiazol tetrazolium (MTT) assay was utilized to test sensitivity of gastric cancer tissues and cancer-adjacent normal tissues to L-OHP. Results The expression level of miR-150 in gastric cancer tissues (0.324 1±0.063 6) was significantly lower than in cancer-adjacent normal tissues (0.576 0±0.148 0) (t=-13.987, P=0.000), and those of MDR1, GST-π, bcl-2 and Survivin mRNAs [(1.217 7±0.272 6), (0.695 3±0.256 7), (0.898 5±0.196 2), (0.826 2±0.270 8), respectively] were all higher than in cancer-adjacent normal tissues [(0.302 0±0.092 1), (0.347 7±0.162 8), (0.195 2±0.085 6), (0.348 6±0.172 0), respectively] (t=28.464, P=0.000; t=10.228, P=0.000; t=29.387, P=0.000; t=13.316, P=0.000). Results of MTT assay showed that relative cell activty to L-OHP was (62.11±25.65)%; cell activity in high-expressed miR-150 group [(52.02±28.71)%] was weaker than in low-expressed miR-150 [(73.27±15.73)%] (t=-4.045, P=0.000); cell activity in positively expressed P-gp, bcl-2, Survivin proteins groups was stronger than in negatively expressed groups, while no association was verified between cell activity (treated with L-OHP) and expression of GST-π (t=0.228, P=0.821). Negative correlation was presented between miR-150 and MDR1, or between miR-150 and bcl-2 (r=-0.571, P=0.000; r=-0.561, P=0.000); while no significant correlations were found between miR-150 and GST-π, or between miR-150 and Survivin (r=0.0882, P=0.437; r=0.186, P=0.098). Conclusion miR-150 might be involved in drug resistance of gastric cancer cells to L-OHP by regulating MDR1, bcl-2 genes. Key words: MicroRNA-150; Gastric cancer; Oxaliplatin; Drug resistance

Effect of microRNA-1 on the expression of angiogenesis-related factors in gastric cancer cells

Objective To observe the role of microRNA (miRNA, miR)-1 in the development of gastric cancer. Methods Real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) was used to detect the expression of miR-1 in human gastric cancer cell lines SGC7901 and BGC823. The plasmid expressing miR-1 was transfected into gastric cancer cells by transient transfection to detect the effect of miR-1 on the proliferation of gastric cancer cells. The effects of over-expressed miR-1 on angiogenesis-related factors vascular endothelial growth factor (VEGF)-A and EDN-1 in gastric cancer cells were detected by Western blotting. Double-reporter assay was used to analyze miR-1 interaction with VEGF-A and EDN1 3’untranslated regions (3’UTR). Results Compared with normal gastric mucosal epithelial cells, miR-1 was down-regulated in gastric cancer cells (P=0.000), and miR-1 inhibited VEGF-A and EDN-1 in proteins levels by binding to specific sites of VEGF-A and EDN-1 3’UTR, thereby inhibiting the proliferation of gastric cancer cells. Conclusion miR-1 inhibits tumors by inhibiting angiogenesis-related factors in human gastric cancer. Key words: Gastric cancer; MicroRNA-1; Vascular endothelial growth factor

Expression and clinical significances of miR-215 and RUNX1 in retinoblastoma

Objective To investigate the expression and clinical significances of miR-215 and runt-related protein1 (RUNX1) in retinoblastoma (RB), and to study the regulation effect of miR-215 on RUNX1 in the retinoblastoma cell line PMC-RB. Methods The expressions of miR-215 and RUNX1 in the tumor tissue, non tumor tissues adjacent to cancer, human RB cell line FMC-RB and human normal retinal vascular endothelial cell line ATCC of RB patients were detected by quantitative real-time polymerase chain reaction (qRT-PCR). miR-215 mimics (miR-215-mimic), miR-NC, si-RUNX1 and si-NC were transfected into FMC-RB cell line respectively. Cell proliferation, migration and invasion ability were measured respectively, thus detecting the regulation effect of miR-215 on RUNX1. Results The expression of miR-215 in RB tissues was significantly lower than that in non tumor tissues adjacent to cancer, while the mRNA expression of RUNX1 was higher than that in non tumor tissues adjacent to cancer (P<0.05). The expression of miR-215 in PMC-RB cells was lower than that in ATCC, while the mRNA expression of RUNX1 was higher than that in ATCC (P<0.05). The expression of miR-215 and RUNX1 in RB tumor tissues were closely related to the clinicopathological features of optic nerve infiltration, tumor tissue differentiation and lymph node metastasis (P<0.05). Cell proliferation, migration and invasion in miR-215-mimic group were significantly lower than those in miR-NC group (P<0.05). In transfected 3′ untranslated region (3′UTR)-Wt cells, the luciferase activity in miR-215-mimic group was lower than that in miR-NC group (P<0.05); the expression level of RUNX1 protein in transfected miR-215-mimic cells was lower than that in transfected miR-NC cells (P<0.05). Cell proliferation, migration and invasion in si-RUNX1 group were all lower than those in si-NC group (P<0.05). There was a negative correlation between mRNA expression level of miR-215 and RUNX1 in RB tumor tissues. Conclusions During the occurrence and development of RB, the down-regulation of miR-215 expression can promote malignant progression of tumor by targeting RUNX1. miR-215 can be used as a biological markers and therapeutic target for RB diagnosis. Key words: Retinoblastoma; MicroRNAs; Core binding factor alpha 2 subunit; Cell line, tumor

Testicular nuclear receptor 4 affects the chemo sensitivity of prostate cancer DU145 cells to docetaxel by regulating microRNA-145 expression

Objective To investigate whether testicular nuclear receptor 4 (TR4) affects the chemo-sensitivity of prostate cancer DU145 cells to docetaxel by regulating microRNA-145 (miR-145) expression. Methods The expression of TR4 was overexpressed or inhibited in prostate cancer DU145 cells, the microRNA-145 (miR-145) status in those cells was evaluated. Luciferase assay was used to detect whether TR4 could affect the activity of miR-145 promoter. Different concentration of docetaxel were added to those DU145 cells which were either inhibited the expression of TR4, miR-145 or both, 48 hours later, cell counting kit-8 (CCK-8) assay was used to detect the cell activity in each group, and the half maximal inhibitory concentration (IC50) were calculated. Results MiR-145 significantly decreased when TR4 is overexpressed and vice versa; The activity of miR-145 promoter was significantly increased when TR4 was suppressed (P=0.016); when TR4 was inhibited, the chemo sensitivity of DU145 to docetaxel were increased with a lower IC50 value (2.602 μg/ml) as compared to that of control group (5.397 μg/ml) (P=0.001). When miR-145 was suppressed, the chemo-sensitivity of DU145 cells to docetaxel were decreased with a higher IC50 value (6.179 μg/ml) as compared to control group (3.549 μg/ml) (P=0.001). When both TR4 and miR-145 were inhibited in DU145 cells, they had a similar IC50 (4.578 μg/ml) as compared to that of the control group (4.122 μg/ml) (P=0.123). Conclusion TR4 could influence the chemo sensitivity of prostate cancer DU145 cells to docetaxe by transcriptionally regulating the expression of miR-145. Key words: Prostate cancer; Testicular nuclear receptor 4; MicroRNA-145; Docetaxel; Half maximal inhibitory concentration

MiR-145 regulates proliferation and chemotherapy sensitivity of ovarian carcinoma

miR-145 regulates proliferation and chemotherapy sensitivity of ovarian carcinoma

Regulation of microRNA-22 on glycometabolism of hematopoietic stem cell TF-1 and its molecular mechanism

Objective To study the regulation of microRNA-22(miR-22) on glycometabolism of hematopoietic stem cell TF-1 and its molecular mechanism. Methods TF-1 cells were cultured for 2 h under hypoxic conditions.The expression levels of Glut4 and miR-22 was detected by quantitative real-time PCR(qRT-PCR). The sgRNA vector of the miR-22 gene was constructed and miR-22 gene of TK-1 cells was knocked out by the CRISPR/Cas9 technique.Overexpression vectors were constructed, and miR-22 knocked-out cells were introduced to overexpress miR-22, the expression of miR-22 was detected by qRT-PCR and the expression levels of Glut4 and PPAR-γ were detected by qRT-PCR and Western blot. Results Compared with the control group, the expression of miR-22 in TF-1 cells decreased (0.015±0.000 vs. 0.056±0.001) and the expression of Glut4 (0.351±0.038 vs. 0.152±0.005) and PPAR-γ (0.421±0.017 vs. 0.248±0.008) increased, when TF-1 cells were cultured for 2 h under hypoxic conditions, and the differences were statistically significant (all P<0.05). Compared with the control group, the expression levels of Glut4 (0.019±0.00 vs. 0.008±0.000) and PPAR-γ (0.038±0.001 vs. 0.019±0.000) were significantly increased after miR-22 gene silencing, and they were significantly decreased (Glut4: 0.005±0.000 vs. 0.008±0.000; PPAR-γ: 0.137±0.000 vs. 0.019±0.000) after overexpression of miR-22, and the differences were statistically significant (all P<0.05). Conclusions It suggests that miR-22 exerts a negative regulation on glycometabolism of hematopoietic stem cell TF-1 by downregulating the expression of PPAR-γ.A new regulatory factor of TF-1 glycometabolism and the mechanisms are identified, which has provided new ideas for the targeted medication of diseases induced by hematopoietic stem cell dysfunction. Key words: Hematopoietic stem cells; Glycometabolism; MicroRNA; Peroxisome proliferators-activated receptor-γ

Expression of microRNA-338 in lung adenocarcinoma and its inhibition mechanism of lung cancer metastasis

Objective To study the expression of microRNA (miRNA, miR)-338 in lung adenocarcinoma cell line and lung adenocarcinoma tissues, and to study the mechanism of lung cancer metastasis. Methods The expression of miR-338 was detected by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) in lung adenocarcinoma cell line A549, and 115 cases of lung adenocarcinoma tissues and normal lung tissues adjacent to tumor. To construct miR-338 over expressing lung cancer cell line and explore the function of miR-338 in the proliferation, invasion and metastasis of lung cancer cells. To detect the expression of integrin β3 in miR-338 over expression lung cancer cell line, and to study the mechanism of miR-338 inhibiting lung cancer metastasis. Results (1)The expression of miR-338 was down regulated in lung adenocarcinoma cell line and lung adenocarcinoma tissues (1.00 vs. 0.15±0.01, P=0.035), (0.00 vs. -2.99±3.69, P=0.001). The expression of miR-338 was correlated with tumor embolus, tumor recurrence, TNM stage (P=0.005, 0.004, 0.025). The 5 years overall survival rate of the low expression group was significantly lower than that of the high expression group (median survival time 44 months vs. 53 months, P=0.001). (2) Up-regulating the expression of miR-338 can inhibit the proliferation of lung cancer cells, migration [average cell count (347±41) cells vs. (190±8) cells, P=0.029] and invasion [average cell count (197±12) cells vs. (68±14) cells, P=0.008]. Xenograft tumor model in nude mice showed that the high expression of miR-338 can effectively inhibit the growth of transplanted tumor [average tumor clone number (20±7) cells vs. (4±2) cells, P=0.004]. (3) In the miR-338 over expression lung adenocarcinoma cell line, the expression of integrin β3 was significantly lower than that in control cells (1.00 vs. 0.35±0.04, P=0.001). Dual luciferase activity assay revealed that the miR-338 over expression group was reduced in the luciferase activity (1.00 vs. 0.75±0.08, P=0.030). Conclusion (1) MiR-338 can inhibit the proliferation, invasion and metastasis of lung cancer cells; (2) MiR-338 play a negative regulation post transcription by targeting the integrin β3 3’untranslated regions (3’UTR) mRNA region. integrin β3 is a new miR-338 lung cancer target gene. Key words: Lung adenocarcinoma; Metastasis; MicroRNA-338; Integrin β3

Expression of microRNA-203 in colorectal cancer cells and its influence on invasion and migration ability

Objective To explore the expression of microRNA (miRNA, miR)-203 in colorectal cancer cell lines, and its influence on invasion and migration ability. Methods Real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) was used to detect the expression level of miR-203. The cells were divided into 3 groups: blank group, miR-203 mimics group and miR-203 inhibitor group according to the cell transfection. Cell migration ability was detected by scratch wound migration assay, and the invasion ability by Transwell assay. Cell adhesion was detected by methyl thiazol tetrazolium (MTT) method. Results Compared to the normal colonic epithelial cells, the expression of miR-203 in colorectal cancer cells were significantly decreased, and the difference was statistically significant (P=0.000, 0.001, 0.005, 0.000). After the transfection of miR-203 mimics, the adhesion ability, invasion [(45.9±6.5) cells] and metastasis ability of colorectal cancer cell lines were significantly inhibited. Conclusion The expression of miR-203 is down-regulated in colorectal cancer, and over-expression of miR-203 can inhibit the invasion and migration ability of colorectal cancer cells. Key words: MicroRNA-203; Colorectal cancer; Invasion; Migration