miR-145-5p affects the proliferation of colorectal cancer cells and cytotoxicity of NK cells by targeting regulation of RAD18 expression / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探索RAD18影响结直肠癌细胞增殖及调节NK细胞对结直肠细胞的杀伤作用及其可能的机制。方法：采用生物信息学技术分析结直肠癌组织中RAD18和miR-145-5p的表达及两者之间的调控关系、分析RAD18富集通路。采用qPCR法验证RAD18和miR-145-5p在结直肠癌细胞中的表达，双荧光素酶报告基因实验验证miR-145-5p与RAD18的调控关系。按转染物的不同将SW480、HCT-15细胞分为将si-RAD18组、si-NC组，另向SW480细胞分别转染inhibitor-NC+si-NC、miR-145-5p inhibitor+si-NC或miR-145-5p inhibitor+si-RAD18，采用CCK-8法、克隆形成实验分别检测敲降miR-145-5p和/或RAD18对细胞增殖、克隆形成的影响；将各组细胞分别与经IL-2激活的NK92细胞共培养，采用乳酸脱氢酶释放法、ELISA和免疫荧光染色法分别检测NK细胞的细胞毒性、细胞因子分泌及细胞表面穿孔素和颗粒酶B表达的影响。结果：RAD18在结直肠癌组织和细胞中呈高表达（均P<0.01）。敲降RAD18可以抑制结直肠癌细胞增殖能力（P<0.05）和促进NK细胞活力、细胞毒性、IFN-γ、TNF-α、GM-CSF分泌及穿孔素和颗粒酶B的表达（均P<0.05）。双荧光素酶报告实验验证了RAD18-3’UTR与miR-145-5p的结合关系，miR-145-5p在结直肠癌组织和细胞中低表达（P<0.05或P<0.01）。miR-145-5p可以靶向下调RAD18的表达（P<0.05），过表达RAD18可以逆转miR-145-5p过表达对NK细胞杀伤效应的促进作用（均P<0.05）。结论：miR-145-5p可靶向下调RAD18的表达，miR-145-5p/RAD18轴能够影响结直肠癌细胞的增殖和NK细胞对其的细胞毒作用。

Biomarker potential of hsa-miR-145-5p in peripheral whole blood of manic bipolar I patients

Objective: Bipolar I disorder (BD-I) is a type of bipolar spectrum disorder characterized by manic or mixed episodes. Detecting microRNA regulations as epigenetic actors in BD-I is important to elucidate the pathogenesis of the disease and reveal the potential of microRNAs (miRNAs) as biomarkers. Methods: We evaluated the expression profile of six candidate miRNAs (hsa-miR-145-5p, hsa-miR-376a-3p, hsa-miR-3680-5p, hsa-miR-4253-5p, hsa-miR-4482-3p, and hsa-miR-4725) in patients with BD-I and in healthy controls (aged 11-50 years). We also determined the potential target genes of these miRNAs through in silico analysis. The diagnostic values of the miRNAs were calculated through receiver operating characteristic curve analysis. Results: Four miRNAs were upregulated (hsa-miR-376a-3p, hsa-miR-3680-5p, hsa-miR-4253-5p, hsa-miR-4482-3p) and hsa-miR-145-5p was downregulated in patients (p < 0.001). The target gene analyses showed that hsa-miR-145-5p specifically targets the dopamine decarboxylase (DDC) gene. The area under the curve of hsa-miR-145-5p was 0.987. Conclusion: Differential expression of five miRNAs in peripheral blood may be associated with the pathogenesis of BD-I, and hsa-miR-145-5p has potential as a BD-I biomarker. This miRNA can be used in dopamine-serotonin regulation and dose adjustment in drug therapy via the DDC gene.

Effects of ginkgo biloba tablets on liver injury in patients with coal-burning-borne arsenism based on DNA damage and repair inhibition / 中华地方病学杂志

Objective:To investigate the role of DNA damage and repair inhibition in the effect of ginkgo biloba on liver injury in patients with coal-burning-borne arsenism.Methods:In March 2017, the investigation was conducted in Jiaole village arsenic poisoning area in Yuzhang Town, Xingren County, Guizhou Province. According to the "Diagnosis of Endemic Arsenicosis" (WS/T 211-2015) and the "Diagnostic Criteria of Occupational Toxic Hepatopathy" (GBZ 59-2010), 52 patients with arsenism were selected as the ginkgo biloba intervention group, and 49 cases of arsenism patients as intervention control group. Ginkgo biloba tablets were given orally for 3 months (1 tablet/time, 3 times/d) according to the commonly used clinical methods, and no other drugs were given to all subjects during the intervention period. The intervention control group was given placebo in the same way as that of ginkgo biloba intervention group. A total of 41 residents who did not burn high arsenic coal 12 km away with no abnormal liver function were selected as normal control group. Physical examinations were performed before the intervention and at the end of the intervention at 3 months. After receiving signed informed consent, morning urine and peripheral venous blood samples were collected to detect urinary arsenic content by inductively coupled plasma mass spectrometry (ICP-MS); liver function biochemical indexes [albumin (ALB), albumin/globulin (A/G), cholinesterase (CHE), total bile acid (TBA)] were determined by automatic biochemical analyzer, DNA damage by single-cell gel electrophoresis assay, and the expression of miR-145 (repair inhibition index) by qRT-PCR.Results:There were 116 subjects, 41 in normal control group, 39 in ginkgo biloba intervention group and 36 in intervention control group. In ginkgo biloba and intervention and intervention control groups, there was no significant difference in age, gender, smoking habits and drinking compared with normal control group ( P > 0.05). Urinary arsenic content, TBA level, DNA damage degree [comet tail DNA percentage (TailDNA%) and olive tail moment (OTM)] and plasma miR-145 expression level [(38.75 ± 19.09) μg/g Cr, (11.13 ± 1.55) μmol/L, 8.50 ± 0.88, 7.43 ± 0.68, 5.78 ± 0.75, respectively] in ginkgo biloba intervention group patients before intervention were higher than those in normal control group [(11.62 ± 5.33) μg/g Cr, (5.36 ± 0.87) μmol/L, 5.24 ± 0.33, 4.71 ± 0.29, 2.05 ± 0.27, respectively], the differences were statistically significant ( P < 0.05); the levels of ALB, A/G and CHE were significantly lower than those in normal control group ( P < 0.05). After the intervention of ginkgo biloba, urinary arsenic content, TBA level, DNA damage degree (TailDNA% and OTM) and plasma miR-145 expression level in patients were significantly lower than those before the intervention ( P < 0.05); the levels of ALB, A/G and CHE were significantly higher than those before the intervention ( P < 0.05). There was no significant difference in the above indexes before and after intervention in the intervention control group ( P > 0.05). The results of correlation analysis between DNA damage degree, miR-145 and liver function indexes after the intervention of ginkgo biloba showed that, DNA damage degree (TailDNA% and OTM) was negatively correlated with the levels of ALB, A/G and CHE ( r = - 0.34, - 0.33, - 0.48, - 0.31, - 0.31, - 0.42, P < 0.05), and positively correlated with the level of TBA ( r = 0.49, 0.48, P < 0.05); miR-145 was negatively correlated with the levels of ALB, A/G and CHE ( r = - 0.26, - 0.23, - 0.38, P < 0.05), which was positively correlated with the level of TBA ( r = 0.32, P < 0.05); and DNA damage degree was positively correlated with the expression of miR-145 ( r = 0.65, 0.52, P < 0.05). Conclusion:Ginkgo biloba tablets can alleviate the liver damage caused by arsenic through coal burning, and the mechanism of this process is related to its inhibition of miR-145 expression and reduction of DNA damage.

Effects of circular RNA circOMA1 on cell proliferation and apoptosis of children with acute myeloid leukemia / 中华实用儿科临床杂志

Objective:To investigate the expression of circular RNA circOMA1 in children with acute myeloid leukemia (AML) and to investigate the effect and mechanism of circOMA1 on the cell proliferation and apoptosis of human acute monocytic leukemia cells (THP-1).Methods:Bone marrow samples of 14 children with AML at the initial diagnosis and after complete remission were collected as the initial diagnosis group and remission group, and bone marrow samples from 10 children without tumor or malignant blood disease in the same hospital and the same period were enrolled as the control group.Real-time quantitative polymerase chain reaction (qPCR) was used to detect the expression of circOMA1, miR-145 and myelocytomatosis viral oncogene homolog (MYC) mRNA in clinical AML samples and THP-1 cell line.The cells transfected with THP-1 were divided into groups, the cells transfected with circOMA1 alone (circOMA1 high expression group) were transfected with pcDNA3.1 empty vector cells as control (pcDNA3.1 control group); the cells co transfected with circOMA1 and miR-145 (circOMA1+ miR-145 group) were treated with pcDNA3.1 and miR-NC co transfected cells were used as control (pcDNA3.1+ miR-NC group, circOMA1+ miR-NC group). Cell counting kit (CCK8) was adopted to detect the effects of circOMA1 and miR-145 on the cell proliferation of THP-1.The effects of circOMA1 and miR-145 on cell apoptosis of THP-1 were detected using flow cytometry, and the effects of circOMA1 and miR-145 on MYC protein expression was detected via Western blot.The comparison between groups was analyzed by independent sample t-test or paired sample t-test, and the correlation was analyzed by Pearson correlation. Results:The expression of circOMA1 in the initial diagnosis group(4.408±3.607) was significantly increased compared with that in the control group (0.998±0.560) ( t=2.946, P<0.01); the expression of circOMA1 in remission group(1.582±0.950) was significantly decreased compared with that in the initial diagnosis group( t= 3.628, P<0.01). The THP-1 cell experiments showed that compared to the pcDNA3.1 control group, the expression of miR-145 in the circOMA1 high expression group decreased ( t= 4.21, P<0.05), cell proliferation was enhanced at 72 h and 96 h ( t=5.46, 7.40, all P<0.05), apoptosis was inhibited( t=6.44, P<0.01). The expression of MYC protein in circOMA1+ miR-NC group was higher than that of pcDNA3.1+ miR-NC group( t=5.72, P<0.01), the expression of MYC protein in circOMA1+ miR-145 group was lower than that in circOMA1+ miR-NC group ( t=4.56, P<0.05); at 72 h and 96 h, the cell proliferation level of circOMA1+ miR-NC group was higher than that of pcDNA3.1+ miR-NC group ( t=5.77, 7.30, all P<0.05), the level of cell proliferation in circOMA1+ miR-145 group was lower than that in circOMA1+ miR-NC group ( t=4.66, 6.17, all P<0.05); the apoptosis rate of circOMA1+ miR-145 group was higher than that of circOMA1+ miR-NC group ( t=4.25, P<0.05). circOMA1 expression was negatively correlated with miR-145 expression ( r=-0.62, P=0.016) and positively correlated with MYC gene expression ( r=0.64, P=0.013) in clinical samples. Conclusions:circOMA1 is highly expressed in children with AML, and can promote AML cell proliferation and inhibit apoptosis through miR-145/MYC pathway, which provides a basis for AML therapy and diagnosis.

Role of miR-145-5p Targeting ARK5 in Regulating the Proliferation and Apoptosis of Human Epithelial Ovarian Cancer Cells / 中国医学科学院学报

Objective To explore the effect of miR-145-5p on the proliferation and apoptosis of human ovarian cancer cells and the possible molecular mechanisms involved.Methods Real-time quantitative PCR was performed to detect the expression of miR-145-5p in ovarian epithelial cells and ovarian cancer cells.CCK-8 and flow cytometry were used to detect the effects of miR-145-5p overexpression on the proliferation and apoptosis of ovarian cancer cells.TargetScan was employed to predict the target genes of miR-145-5p.Western blotting,dual luciferase reporter assay and rescue experiment were employed to predict and verify the underlying molecular mechanism of miR-145-5p function.Results The expression of miR-145-5p in ovarian cancer cells was significantly lower than that in normal ovarian epithelial cells(

Study on the effects and mechanism of Linc01278 on the biological behavior of prostate cancer cells / 中华内分泌外科杂志

Objective:To investigate the effect and molecular mechanism of long non-coding RNA (Linc01278) on the proliferation, invasion and migration, and tumor phenotype of prostate cancer cells.Methods:Prostate cancer tissues and corresponding normal tissues adjacent to the cancer were obtained in 46 patients with prostate cancer confirmed by the hospital from Dec. 2018 to Dec. 2019. Real-time fluorescence quantitative PCR (qPCR) was used to detect the expression levels of Linc01278 and miR-145 in prostate cancer tissues and adjacent normal tissues. The prostate cancer PC-3 cells were transfected with plasmids and cells were divided into four groups: blank group, control group, overexpression group and rescue group. The blank group was not given any treatment; the control group was transfected with blank control vector and miRNA Control; the overexpression group was transfected with Linc01278 overexpression vector and miRNA Control; the rescue group was transfected with Linc01278 overexpression vector and miR-145 mimics. Bioinformatics analysis was used to predict the binding site of Linc01278 to miR-145. Dual Luciferase Reporter Gene Assay was used to analyze the adsorption of Linc01278 on miR-145; Transwell experiment was used to detect the invasion and migration ability of the four groups of cells; CCK-8 method was used to detect the cell proliferation ability of the four groups of cells. Western blot was used to detect the expression of OCT4 and SOX2 in the four groups of cells; qPCR was used to detect the expression of OCT4, SOX2 and miR-145 in the four groups of cells.Results:Compared with adjacent normal tissues, Linc01278 was significantly higher in prostate cancer tissues (adjacent normal tissues: 0.012±0.002 vs prostate cancer tissues: 0.022±0.002, P=0.0072) , while miR-145 was significantly lower (adjacent normal tissues: 0.117±0.011 vs prostate cancer tissues: 0.081±0.007, P=0.0005) . Correlation of both was negative significantly ( P=0.0012) . Targetscan analysis revealed that the 804-825 position of the Linc01278 transcript had a complementary base pairing with miR-145. The results of the double luciferase reporter gene showed that miR-145 mimics could significantly reduce the luciferase activity of Linc01278 ( P<0.01) . Compared with the blank group and the control group, the invasion and migration cells, the relative proliferation ability, the expression of Oct4 and Sox2 mRNA and protein were significantly increased, and the expression of miR-145 was significantly decreased in the overexpression group ( P<0.05) . While compared with the overexpression group, the invasion and migration cells, the relative proliferation ability, Oct4 and Sox2 were significantly decreased in the rescue group, and the expression of miR-145 was significantly increased ( P<0.01) . Conclusion:Linc01278 may promote the proliferation, invasion and migration of prostate cancer cells by specifically adsorbing miR-145.

Purple sweet potato anthocyanin regulates the proliferation, migration and invasion of breast cancer MDA-MB-231 cells through circ_0003998/miR-145 axis / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨紫甘薯花色苷（purple sweet potato anthocyanin, PSPA）是否通过circ_0003998/miR-145轴调控乳腺癌MDA-MB-231细胞的增殖、迁移和侵袭。方法：选用乳腺癌MDA-MB-231细胞，将其分为对照组，200、400和800 μg/ml PSPA组，pcDNA组、pcDNA-circ_0003998组、si-NC组、si-circ_0003998组、si-circ_0003998+anti-miR-145组、PSPA+pcDNA组、PSPA+pcDNA-circ_0003998组和PSPA+anti-miR-145组。用qPCR法检测细胞中circ_0003998和miR-145的表达，CCK-8法、Transwell小室法分别检测转染前后细胞的增殖、迁移和侵袭能力，WB法检测细胞中Ki-67、MMP-2和MMP-9蛋白的表达。用双荧光素酶报告基因实验验证circ_0003998与miR-145的靶向关系。结果：与对照组比较，各剂量PSPA组MDA-MB-231细胞的增殖抑制率、miR-145表达水平均显著升高（均P<0.01），Ki-67、MMP-2、MMP-9蛋白和circ_0003998的表达水平、细胞迁移和侵袭细胞数均显著降低（均P<0.01），并呈现浓度依赖性。circ_0003998可以靶向负调控miR-145的表达。敲减circ_0003998后，MDA-MB-231细胞的增殖抑制率、miR-145表达水平显著升高，Ki-67、MMP-2和MMP-9蛋白表达水平、细胞迁移和侵袭细胞数均显著减少（均P<0.01）。共转染si-circ_0003998和anti-miR-145则可逆转敲减circ_0003998表达对MDA-MB-231细胞增殖、迁移和侵袭的抑制作用，过表达circ_0003998或抑制miR-145表达可逆转PSPA对MDA-MB-231细胞增殖、迁移和侵袭的抑制作用。结论：PSPA通过circ_0003998/miR-145轴抑制乳腺癌MDA-MB-231细胞的增殖、迁移和侵袭。

circRNA_001569 affects the malignant biological behavior of breast cancer cells through the miR-145/HBXIP axis / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨circRNA_001569通过miR-145/HBXIP轴在乳腺癌细胞增殖、侵袭、迁移中发挥的作用。方法:收集2016年1月至2019年1月期间衡水市人民医院收治的30例乳腺癌患者的癌组织和癌旁组织。qPCR检测circRNA_001569在乳腺癌组织、癌旁组织以及细胞系中的表达。生物信息学工具预测miR-145的靶基因，RNA免疫沉淀（RNA immunoprecipitation，RIP）和双荧光素酶报告基因实验检测miR-145或靶基因之间的相互作用；向乳腺癌MDA-MB-231和MCF-7细胞中转染si-circRNA_001569、miR-145 mimics或miR-145 inhibitor，建立基因过表达或沉默的细胞模型，qPCR和Western blotting分别检测转染对相关基因和蛋白表达的影响，CCK-8法、Transwell实验检测转染对细胞增殖、侵袭和迁移的影响。结果:在乳腺癌组织和乳腺癌细胞中，circRNA_001569和HBXIP均呈高表达、miR-145呈低表达。RIP分析和双荧光素酶实验证实了miR-145与circRNA_001569和HBXIP之间的靶向关系；circRNA_001569或HBXIP过表达促进MDA-MB-231和MCF-7细胞的增殖、侵袭和迁移（均P<0.01），而miR-145过表达起相反的作用（均P<0.01）。结论:circRNA_001569可能通过下调miR-145的表达、上调HBXIP的表达从而促进乳腺癌细胞的增殖、侵袭和迁移。

Over-expression of miR-145-5p inhibits malignant biological behaviors of esophageal squamous cells carcinoma via down-regulating IGF1R / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To explore the mechanism of miR-145-5p on malignant biological behaviors, such as pro-liferation, invasion, migration and epithelial-mesenchymal transition (EMT), of esophageal squamous cell carcinoma (ESCC) TE-10 cells. Methods: The expression of miR-145-5p in ESCC cell lines and normal cells was detected by PCR. Dual luciferase reporter gene assay was used to detect the targeted regulation between miR-145-5p and insulin-like growth factor 1 receptor (IGF1R). The expres-sions of IGF1R protein and EMT related proteins were detected by Western blotting. Transwell assay and CCK-8 assay were carried out to detect the effects of miR-145-5p/IGF1R axis on the proliferation, migration andinvasionofTE-10 cells. Results: miR-145-5p was down-regulated in ESCC cell lines with the lowest expression in TE-10 cells (P<0.01orP<0.05).Over-expression of miR-145-5p significantly inhibited proliferation, invasion, migration and EMT of TE-10 cells (P<0.01 or P<0.05). Dual luciferase reporter gene assay con-firmed that miR-145-5p targetedly down-regulated IGF1R expression (P<0.01). The restora-tion experiments further confirmed that simultaneous over-expression of miR-145-5p and IGF1R significantly attenuated the promotion effect of IGF1R on proliferation, invasion, migration and EMT of TE-10 cells (P<0.01 or P<0.05). Conclusions: Over-expression of miR-145-5p inhibits proliferation, invasion, migration and EMT of ESCC TE-10 cells by down-regulating IGF1R.

Regulatory Mechanism of MicroRNA-145 in the Pathogenesis of Acute Aortic Dissection

PURPOSE: Previous studies have confirmed that microRNAs play important roles in the pathogenesis of acute aortic dissection (AAD). Here, we aimed to explore the role of miR-145 and its regulatory mechanism in the pathogenesis of AAD. MATERIALS AND METHODS: AAD tissue samples were harvested from patients with aortic dissection and normal donors. Rat aortic vascular smooth muscle cells (VSMCs) were transfected with miR-145 mimic/inhibitor or negative control mimic/inhibitor. Gene and protein expression was measured in human aortic dissection tissue specimens and VSMCs by qRT-PCR and Western blot. Luciferase reporter assay was applied to verify whether connective tissue growth factor (CTGF) was a direct target of miR-145 in VSMCs. Methyl thiazolyl tetrazolium assay was used to detect VSMC viability. RESULTS: miR-145 expression was downregulated in aortic dissection tissues and was associated with the survival of patients with AAD. Overexpression of miR-145 promoted VSMC proliferation and inhibited cell apoptosis. Moreover, CTGF, which was increased in aortic dissection tissues, was decreased by miR-145 mimic and increased by miR-145 inhibitor. Furthermore, CTGF was confirmed as a target of miR-145 and could reverse the promotion effect of miR-145 on the progression of AAD. CONCLUSION: miR-145 suppressed the progression of AAD by targeting CTGF, suggesting that a miR-145/CTGF axis may provide a potential therapeutic target for AAD.

LncRNA MALAT1/MiR-145 Adjusts IL-1β-Induced Chondrocytes Viability and Cartilage Matrix Degradation by Regulating ADAMTS5 in Human Osteoarthritis

PURPOSE: Accumulating evidence suggests that microRNA-145 (miR-145) plays an important role in osteoarthritis (OA), which is a chronic progressive joint disease. Long noncoding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) promotes metastasis in cancers and functions as a sponge for miR-145. However, the role of MALAT1/miR-145 in OA pathogenesis has not yet been elucidated. MATERIALS AND METHODS: The expression of MALAT1 and miR-145 was examined by quantitative real-time PCR; the interaction between miR-145, MALAT1 and a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) 5 was verified by luciferase reporter assay. Correlations among MALAT1, miR-145, and ADAMTS5 were analyzed by Spearman rank analysis. Chondrocytes viability and cartilage extracellular matrix (ECM) degradation were investigated with cell viability assay and Western blotting analyzing expression of ADAMTS5, collagen type 2 alpha 1 (COL2A1), aggrecan (ACAN), and cartilage oligomeric matrix protein (COMP). RESULTS: MALAT1 was upregulated, and miR-145 was downregulated in OA samples and IL-1β-induced chondrocytes. Mechanically, miR-145 could directly bind to MALAT1 and ADAMTS5. Moreover, miR-145 expression was negatively correlated with MALAT1 and ADAMTS5 expression in OA patients, whereas MALAT1 and ADAMTS5 expression was positively correlated. Functionally, overexpression of MALAT1 inhibited chondrocyte viability and promoted cartilage ECM degradation in IL-1β-induced chondrocytes. In support thereof, MALAT1 silencing and miR-145 upregulation exerted the opposite effect in IL-1β-induced chondrocytes. Moreover, the effect of MALAT1 was counteracted by miR-145 upregulation, and ADAMTS5 restoration could abate miR-145 effects. CONCLUSION: An MALAT1/miR-145 axis contributes to ECM degradation in IL-1β-induced chondrocytes through targeting ADAMTS5, suggesting that MALAT1/miR-145/ADAMTS5 signaling may underlie human OA pathogenesis.

Effect of long-chain non-coding RNA TUG1 on radiosensitivity of human cervical cancer XB1702 cells by adsorption of miR-145 / 中华放射肿瘤学杂志

Objective@#To evaluate the effect of long-chain non-coding RNA TUG1 on the radiosensitivity of cervical cancer cells and explore its underlying mechanism.@*Methods@#The expression of TUG1 and miR-145 in cervical cancer cells XB1702 and normal endometrial stromal cells (ESCs) was detected by qRT-PCR. The transfected si-NC, transfected si-TUG1, transfected si-NC combined with irradiation, transfected si-TUG1 combined with irradiation, si-TUG1 and anti-miR-NC co-transfected and, si-TUG1 and anti-miR-145 co-transfected groups were established, which were transfected into XB1702 cells by liposome method. The survival fraction of each group was detected by colony formation assay. The cell apoptosis of each group was detected by flow cytometry. The fluorescence activity of each group was assessed by dual luciferase reporter gene assay.@*Results@#Compared with the normal ESCs, the expression of TUG1 was significantly up-regulated, whereas that of miR-145 was significantly down-regulated in the cervical cancer cells XB1702. Silencing TUG1 significantly increased the survival fraction of XB1702 cells, promoted cell apoptosis and enhanced the radiosensitivity of irradiation to XB1702 cells. TUG1 could target and regulate the expression of miR-145. Suppressing miR-145 reversed the silencing effect of TUG1 on inhibiting proliferation, accelerating apoptosis promotion and enhancing sensitization of XB1702 cells.@*Conclusions@#Silencing long-chain non-coding RNA TUG1 can enhance the radiosensitivity of cervical cancer cells. The mechanism may be related to targeting miR-145, which will provide a target for radiotherapy of cervical cancer.

Novel cancer-specific nanoparticles mediated photodynamic therapy inhibits human cervical cancer cells in vitro / 西安交通大学学报(医学版)

Objective: To determine the effects of photodynamic therapy (PDT) with R11-SSPEI/miR-145 as photosensitizer on cell death in cervical cancer so as to investigate the underlying mechanisms. Methods: Cell cytotoxic effect was measured by MTT assay. Cellular apoptosis was detected by AnnexinV-PI by flow cytometry. The level of reactive oxygen species (ROS) was observed by DCFH-DA staining. The expressions of Caspase-3, Caspase-9, BIP, and Bcl-2 were assessed by Western blotting. Mitochondrial membrane potential was tested by JC-1 staining. Results: Analysis of cell proliferation evidenced that there was a dramatic inhibition after photodynamictherapy (PDT) with R11-SSPEI/miR-145. We observed increased apoptosis (P=0.014) and demonstrated that the apoptosis was involved of mitochondrial and endoplasmic reticulum death pathway. The expressions of Caspase-3, Caspase-9 and BIP were elevated (P<0.05) and that of Bcl-2 decreased. NAC could decrease the ROS level in photodynamic therapy with R11-SSPEI/miR-145 group and rescue the cell death. Conclusion: Taken together, the present results indicated that PDT with R11-SSPEI/miR-145 suppressed cancer development with Hela cells, suggesting R11-SSPEI/miR-145 as a new photosensitizer in PDT has the potential to become the mainstream of cancer treatment.

Effect of miR-145 on Migration and Invasion of Ovarian Cancer Cells / 中国医学科学院学报

Objective To evaluate the effect of miR-145 on migration and invasion of ovarian cancer cells.Methods The effect of miR-145 overexpression on the expression levels of miR-145 and zeb-2 were detected with qRT-PCR and Western blotting.The changes of migration and invasion were examined using Transwell assay.Target genes of miR-145 were predicted by bioinformatics software.Dual-luciferase reporter assay were used to verify zeb-2 as a direct target of miR-145.zeb-2 siRNA was transiently transfected in SKOV3 and 3AO cells,Transwell was used to examine migration and invasion abilities.Results The migration and proliferation of SKOV3(=10.752,=0.000;=5.617,=0.005)and 3AO cells(=10.111,=0.001;=21.746,=0.000)decreased significantly after overexpression of miR-145.The results of dual-luciferase reporter assay showed that the relative luciferase activity of co-transfected miR-145 mimic and WT 3'UTR expression vectors was significantly lower than that of co-transfected mimic control and WT 3'UTR expression vectors(SKOV3:=4.572,=0.010;3AO:=3.528,=0.024).There was no significant difference in relative luciferase activity between co-transfected miR-145 mimic/MUT 3'UTR expression vector cells and co-transfected mimic control/MUT 3'UTR expression vector cells(SKOV3:=0.227,=0.831;3AO:=0.040,=0.970).Real-time quantitative PCR showed that the zeb-2 expressions in SKOV3(=1.490,=0.211)and 3AO cells(=0.114,=0.914)were not significantly different from negative control after 48 h of miR-145 overexpression.Western blot analysis showed that the expression of zeb-2 protein in SKOV3(=3.769,=0.020)and 3AO cells(=4.452,=0.011)decreased significantly compared with negative control after 72 h of miR-145 overexpression.Seventy-two hours after transfection of zeb-2 siRNA,Western blotting showed that the expression of zeb-2 protein in SKOV3(=4.660,=0.010)and 3AO cells(=4.594,=0.010)was significantly down-regulated.Transwell assay showed that the migration and invasion abilities of SKOV3(=18.655,=0.000;=18.026,=0.000)and 3AO cells(=5.500,=0.005;=8.780,=0.001)were significantly decreased.Conclusion miR-145 may inhibit the migration and invasion of ovarian cancer cells by targeting zeb-2.

Reversal of miR-145 in triple-negative breast cancer by estrogen and progesterone combined administration / 安徽医科大学学报

Objective To investigate the role of estrogen and progesterone in the tumorigenesis of triple-negative breast cancer and its molecular mechanism. Methods To construct of MDA-MB-231/miR-145 and MDA-MB-231/ miR-SCR(scramble control) stable cell lines. The cells were cultured in five groups that were respectively miR-SCR group, miR-145 group, miR-145 + E2 group, miR-145 + P4 group, miR-145 +E2 +P4 group, grouped adminis-tradion. Cell proliferation was detected by CCK-8 kit, the expression level of miR-145 target gene insulin-like growth factor receptor-1 (IGF-IR), N-RAS was detected by Western blot, expression of IGF-1R, N-RAS downstream signaling molecule vascular endothelial growth factor(VEGF) and expression of miR-145 after transfection was detected by real-time quantitative PCR(qRT-PCR). Results The results of CCK-8 assay showed that estrogen and progesterone combined administration could reverse the inhibitory effect of miR-145 on the proliferation of triple-negative breast cancer cells (P ＜ 0. 05 ). The results of Western blot showed that estrogen and progesterone combined administration could increase the expression level of IGF-1 R and N-RAS in the target gene of miR-145. The results of qRT-PCR showed that the expression level of miR-145 was significantly higher than that of control group (P ＜ 0. 05) and the expression of VEGF was significantly increased by estrogen and progesterone combined administration (P ＜0. 01). Conclusion Estrogen and progesterone combined administration can promote the expression of VEGF and cell proliferation, and then reverse the anti-tumor effect of miR-145 in the triple-negative breast cancer.

Effect of miR-145 on diabetic neuropathic pain by down-regulating Nav1.8 / 临床麻醉学杂志

Objective To observe the effect of miR-145 on pain threshold and explore the pos-sible underlying positive role of miR-145 in rats with diabetic neuropathic pain.Methods The total of 36 rats with diabetic neuropathic pain were randomly divided into three groups respectively with nor-mal control group (group N)(n=12 for each group):diabetic neuropathic pain(DNP)group (group D),DNP-NC group (group DN)and DNP-agomiR-145 group (group agomiR-145).The rats received agomiR-145 intrathecal injection in group agomiR-145 (10 μl,1×106TU/ml),or the negative control virus in group DN (10 μl,1×106TU/ml),or equal volume of normal saline in other two groups. Paw mechanical withdrawal threshold(MWT)and paw withdrawal latency(TWL)were measured on the day before intrathecal injection and day 1,days 3,7 and 14 after intrathecal injection.On the days 14 after pain-related behavioral test,the RNA expression of miR-145 in the dorsal root ganglion (DRG)was detected using reverse transcription-quantitative polymerase chain reaction(RT-PCR)as-say and the expression of Nav 1.8 in DRG were detected by fluorescent immunofluorescence.In addi-tion,a dual luciferase activity assay was used to testify the target genes of miR-145.Results MWT and TWL were decreased at 1 d before intrathecal injectionin groups D,DN and agomiR-145 than that in group N (P<0.05).The significant increase of MWT was observed in group agomiR-145 on day 3,7,14 than those in group D and group DN (P<0.05).TWL in group agomiR-145 was increased significantly on day 7 and day 14 compared with those in groups D and DN (P<0.05).Compared with group N,miR-145 expression level in DRG in groups D and DN were significantly lower (P<0.05).In addition,the protein expression of Nav1.8 was significantly increased in group D and DN compared with that in group N (P<0.05).Compared with groups D and DN,miR-145 expression was increased significantly and the expression of Nav1.8 in DRG was decreased significantly in group agomiR-145 (P<0.05).In addition,a dual luciferase reporter assay demonstrated that miR-145 can bind with the 3'-UTR region of Nav1.8 and regulate its expression.Conclusion Intrathecal agomiR-145 can effectively attenuate neuropathic pain of DNP rats,which may be related with down-regulation of Nav1.8 in DRG..

miR-145 downregulates MMP2 expression in lung adenocarcinoma cell A549 / 临床检验杂志

Objective To investigate the regulatory effects of miR-145 on MMP2 expression in lung adenocarcinoma cell A549 and the mechanism involved in the regulation.Methods Lipofectamine 2000 reagent-mediated miR-145 mimic or mimic negative control (NC) was transiently transfected into lung adenocarcinoma cell A549.The expression levels of miR-145 were determined by RT-PCR.MMP2 and β-catenin expression was detected by western blot.Lithium chloride (LiC1),the activator of Wnt/β-catenin pathway,and miR-145 mimic co-processed to act on A549 cells,then the expressions of MMP2 and β-catenin were detected by Western blot.Results The results of RT-PCR showed that the difference of miR-145 in miR-145 mimic group,NC group and blank group was statistically significant(F =296.30,P ＜0.05).western blot showed that the expression levels of MMP2 and nucleus β-catenin were significantly downregulated in miR-145 overexpression group compared with the control (q =8.98,26.59,all P ＜ 0.05).Additionally,the effects of co-processing factors on the cells showed that the activation of wnt/β-catenin pathway reversed the inhibition of miR-145 on MMP2 expression(q =49.47,P ＜ 0.05).Conclusion miR-145 may downregulate MMP2 expression through inhibition of Wnt/β-catenin pathway activity in lung adenocarcinoma cell A549.

The expression and function of miR-145 in papillary thyroid carcinoma / 中华内分泌外科杂志

Objective To study the expression and function of miR-145 in papillary thyroid carcinoma (PTC).Methods PTC tissues and adjacent tissues were collected from 43 cases.Expression of miR-145 in PTC tissues and adjacent tissues was detected with RT-PCR.miR-145 analogue was used to transfect TPC-1cell to upregulate miR-145 expression.Brdu-ELISA method was used to detect the proliferation of TPC-1 cell.Flow cytometry instrument was used to detect the apoptosis and cell cycle of TPC-1 cell.Results RT-PCR test showed that relative expression of miR-145 in thyroid carcinoma tissue was 0.369±0.082,significantly lower than 1.029-±0.365 in tissue adjacent,and the difference was statistically significant (t=3.129,P=0.000).The expression of miR-145 in patients whose biggest tumor size ≥ 1 cm was lower than patients whose biggest tumor size ＜1 cm.Compared with patients with single tumor,the expression of miR-145 in patients with multiple tumor was lower,and the dif ferences were statistically significant (P＜0.05).miR-145 expression was enhanced by miR-145 analogue.Compared with negative control,the proliferative ability of thyroid cancer cell TPC-1 was suppressed significantly,and the difference was statistically significant (P＜0.05).In addition,up-regulation of miR-145 expression could block thyroid cancer TPC-1 in G2/M phase.The apoptosis rate of thyroid cancer cell TPC-1 increased significantly (P＜0.05).Conclusions miR-145 expression is decreased significantly in PTC tissue,and is associated with clinical pathological features.Up-regulation of miR-145 expression can inhibit thyroid cancer cell proliferation,block the cell cycle,and promote apoptosis,miR-145 may play an important role in occurrence and development of thyroid cancer.

Effects of both high glucose and high insulin on proliferations and migrations of human vascular smooth muscle cells as well as miR-145 level / 基础医学与临床

Objective To identify the effect of both high glucose and high insulin on miR-145 level.Methods The human vascular smooth muscle cells ( VSMCs) were cultured and the proliferation of VSMCs was induced by high glucose and high insulin medium.The samples were divided into 4 groups: normal group, high glu-cose group (25 mmol/L), high insulin group (300 mU/L), high glucose and high insulin group.The ex-pression of miR-145 in VSMCs was assayed by real-time PCR.Proliferation of VSMCs was determined by MTT method After 72 h cultivation.The migration of VSMCs was analyzed by cell scratch test .VSMCs in each group was transfected by miR-145 virus ( lentiviral vector ) .Proliferation and migration were assayed after 48 h transfection.Results The expression of miR-145 in VSMCs of other three groups was decreased ( P<0.05 ) , especially the expression in the high glucose and insulin group was the lowest ( P<0.01 ) .Prolifera-tion and migration of VSMCs was promoted by high glucose and/or high insulin medium.Under fluorescent, transfection rate of VSMCs was about 80%after 48 h transfection.Proliferation and migration of VSMCs in each group after transfection were significantly lower than before ( P<0.05 ) .Conclusions High glucose and high in-sulin could decrease the expression of miR-145 in VSMCs, The overexpression of miR-145 may inhibit the prolif-eration and migration of VSMCs .

The effect of microRNA-145 on immune-inflammatory response of foam cells by targeting CD40 / 天津医药

Objective To investigate the effect of miRNA-145 (miR-145) on immuno-inflammatory reaction of foam cells by targeting CD40. Methods Mouse macrophage cell line RAW 264.7 cells cultured in vitro were randomly divided into model group (non-transfected), miR-145 mimics group (transfected miR-145 mimics), miR-145 inhibitor group (transfected miR-145 inhibitor) and silencing CD40 sequence group (transfected siCD40). Then oxidized low density lipoprotein (ox-LDL) was used to stimulate for 24 h to establish immune inflammatory damage cell model. Quantitative real-time polymerase chain reaction (RT-qPCR) and Western blot assay were used to detect the levels of CD 40 mRNA and protein of each group. ELISA was used to detect the levels of inflammatory factors interleukin (IL)-1, IL-6 and tumor necrosis factor (TNF) -α in cell supernatant. Results Compared with model group, the levels of CD40 mRNA, CD40 protein and IL-1, IL-6, TNF-αwere all significantly decreased in miR-145 mimics group (P<0.01). After transfected with miR-145 inhibitor, the above indexes were all significantly increased than those of model group and miR-145 mimics group (P<0.01). After transfected with CD40 siRNA, the levels of CD40 mRNA, CD40 protein and IL-1, IL-6, TNF-αwere all obviously decreased compared with those of miR-145 inhibitor group (P<0.01). Conclusion MiR-145 can regulate the immune inflammatory process of foam cells through the target gene CD40, inhibit the activation of CD40/CD40L signaling pathway and inhibit inflammatory response.