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OBJECTIVE To explore the effect and mechanism of the alcoholic extract from Scabiosa comosa against hepatic fibrosis （HF）. METHODS Intragastrical administration of carbon tetrachloride was given to induce HF model. By observing the pathological changes in liver tissue， mRNA and protein expressions of HF indexes [α-smooth muscle actin （α-SMA）， collagen type Ⅰ] and phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt） pathway-related factors were detected， and the improvement effects and possible mechanism of low-dose， medium-dose and high-dose （50， 100， 200 mg/kg） of alcoholic extract from S. comosa on HF model rats were investigated. Drug-containing serum was prepared by intragastrical administration of alcoholic extract from S. comosa at a concentration of 1 800 mg/（kg·d） （calculated by the amount of raw material）. The effects of drug- containing serum of alcoholic extract from S. comosa on the expression of miRNA-21 were observed through the intervention of HSC-T6 cells with low， medium and high concentrations of drug-containing serum of alcoholic extract from S. comosa （diluted to 10%， 15%， 20%）. miRNA-21 mimics or inhibitors were used to transfect HSC-T6 cells， and the mRNA and protein expressions of factors related to the PI3K/Akt signaling pathway were detected. RESULTS The results of in vivo experiments showed that low， medium and high doses of alcoholic extract from S. comosa significantly ameliorated the histopathological changes in liver tissue of HF rats， and the percentage of collagen was significantly reduced （P＜0.01）； mRNA and protein expressions of the indicators related to HF as well as PI3K and Akt were significantly reduced （P＜0.01）， and mRNA and protein expressions of phosphatase and tensin homolog deleted on chromosome ten （PTEN） were increased in liver tissue of rats （P＜0.01）. The results of in vitro experiments showed that drug-containing serum of alcoholic extract from S. comosa significantly inhibited the expression of miRNA-21 at low， medium and high concentrations （P＜0.01）； whereas after transfection with miRNA-21 mimics， it was found that miRNA-21 mimics significantly increased mRNA and protein expressions of PI3K and Akt （P＜0.01）， while significantly decreased mRNA and protein expressions of PTEN （P＜0.01）； after transfection with miRNA-21 inhibitor， the changes of above indexes were opposite to the above results （P＜0.01）. CONCLUSIONS Alcoholic extracts of S. comosa may inhibit the PI3K/Akt signaling pathway by affecting the expression of miRNA-21， so as to achieve the effect of anti-hepatic fibrosis.
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Objective:To investigate the effect of infliximab combined with miRNA-21 on lung cancer A549 cells.Methods:A549 cells were cultured in vitro and then divided into four groups (blank group, infliximab group, miRNA-21 inhibitor group and combined treatment group) ; CCK-8 test was used to detect cell proliferation; Flow cytometry experiments was employed to detect apoptosis; Western blot was used to detect protein expression.Results:The survival rates of A549 cells in the miRNA-21 inhibitor group and the combined treatment group were 48.67%±2.83% and 25.69%±1.98%, which were significantly different ( P<0.001) ; The proportion of A549 apoptotic cells in the miRNA-21 inhibitor group and the combined treatment group were 46.73%±2.18% and 76.58%±3.67%, respectively, with significant differences ( P<0.001) ; The expression of Caspase-3 (1.21±0.26 vs 0.57±0.07) and Bad (1.08±0.11 vs 0.52±0.06) in the combined treatment group was significantly higher than that of the miRNA-21 inhibitor group in the detection of apoptosis-related proteins, and the expression of Bcl-2 was significantly reduced, with a significant difference ( P<0.001). In the combined treatment group, the expression levels of TNF-α (0.63±0.11 vs 1.23±0. 22, 1.18±0.17, 1.14±0.17) and NF-κB p65 (0.34±0.08 vs 1.31±0.09, 1.29±0.12, 1.11±0.06) were both reduced, and there was a significant difference compared with the other three groups ( P<0.001) . Conclusion:Infliximab combined with miRNA-21 inhibitors can play a synergistic role in lung cancer cells, inhibit the TNF-α/NF-κB signaling pathway, regulate the expression of the Bcl-2 family and Caspase-3, and promote apoptosis, thereby inhibiting lung cancer A549 cell proliferation.
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MicroRNA-21(miRNA-21)is highly expressed in various tumors.Small-molecule inhibition of miRNA-21 is considered to be an attractive novel cancer therapeutic strategy.In this study,fluoroquinolone de-rivatives Al-A43 were synthesized and used as miRNA-21 inhibitors.Compound A36 showed the most potent inhibitory activity and specificity for miRNA-21 in a dual-luciferase reporter assay in HeLa cells.Compound A36 significantly reduced the expression of mature miRNA-21 and increased the protein expression of miRNA-21 target genes,including programmed cell death protein 4(PDCD4)and phos-phatase and tensin homology deleted on chromosome ten(PTEN),at 10 uM in HeLa cells.The Cell Counting Kit-8 assay(CCK-8)was used to evaluate the antiproliferative activity of A36;the results showed that the IC50 value range of A36 against six tumor cell lines was between 1.76 and 13.0 μM.Meanwhile,A36 did not display cytotoxicity in BEAS-2B cells(lung epithelial cells from a healthy human donor).Furthermore,A36 significantly induced apoptosis,arrested cells at the G0/G1 phase,and inhibited cell-colony formation in HeLa cells.In addition,mRNA deep sequencing showed that treatment with A36 could generate 171 dysregulated mRNAs in HeLa cells,while the expression of miRNA-21 target gene dual-specificity phosphatase 5(DUSP5)was significantly upregulated at both the mRNA and protein levels.Collectively,these findings demonstrated that A36 is a novel miRNA-21 inhibitor.
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Objective:To correlate the expression of microRNA (miRNA)-21 and miRNA-335-5p with pepsinogen and gastrin-17 in patients with gastric cancer.Methods:Sixty-one patients with gastric cancer who received treatment in Linhai Second People's Hospital between January 2019 and January 2021 were included in the patient group. An additional 60 healthy patients who concurrently received physical examination were included in the control group. Serum pepsinogen I, pepsinogen II and gastrin-17 levels were determined by latex-enhanced immunoturbidimetry. miRNA-21 and miRNA-335-5p expression were determined by quantitative reverse transcription-polymerase chain reaction. Serum pepsinogen I, pepsinogen II and gastrin-17 levels and miRNA-21 and miRNA-335-5p expression were compared between the two groups. The sensitivity and specificity of miRNA-21 and miRNA-335-5p in the diagnosis of gastric cancer were analyzed. The correlation between miRNA-21 and miRNA-335-5p expression and pepsinogen I, pepsinogen II and gastrin-17 levels was analyzed.Results:Serum pepsinogen I and gastrin-17 levels in the patient group were (54.36 ± 9.89) μg/L and (13.74 ± 1.89) pg/mL, respectively, which were significantly lower than those in the control group [(112.31 ± 23.24) μg/L, (18.75 ± 2.36) pg/mL, t = 17.89, 12.90, both P < 0.05]. Serum pepsinogen II level in the patient group was significantly higher than that in the control group [(24.35 ± 4.53) μg/L vs. (20.37 ± 3.28) μg/L, t = 5.52, P < 0.05]. The relative mRNA expression of miRNA-21 in the patient group was significantly higher than that in the control group [(3.42 ± 0.61) vs. (0.53 ± 0.12), t = 30.01, P < 0.05]. The relative mRNA expression of miRNA-335-5p in the patient group was significantly lower than that in the control group [(0.32 ± 0.17) vs. (1.65 ± 0.35), t = 26.65, P < 0.05]. The receiver operating characteristic curve analysis showed that the sensitivity and specificity of miRNA-21 in the diagnosis of gastric cancer were 74.36% and 68.18%, respectively, and they were 79.49% and 60.90% for miRNA-335-5p, respectively. There was a negative linear correlation between miRNA-21 and pepsinogen I and gastrin-17 levels ( r = -0.82, -0.74), but there was a positive linear correlation between miRNA-21 and pepsinogen II levels ( r = 0.76). There was a positive linear correlation between miRNA-335-5p and pepsinogen I and gastrin-17 ( r = 0.79, 0.72), but there was a negative linear correlation between miRNA-335-5p and pepsinogen II levels ( r = -0.70). Conclusion:miRNA-21 is highly expressed in patients with gastric cancer, while miRNA-335-5p is lowly expressed. miRNA-21 and miRNA-335-5p are highly correlated with pepsinogen and gastrin-17 levels. miRNA-21 and miRNA-335-5p can be used as effective indices for diagnosis of gastric cancer. Findings of this study are highly innovative and scientific.
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Objective:To investigate the molecular mechanism of Qiyu Sanlong prescription （QYSL） in inhibiting the "addiction" of lung cancer A549 cells to miRNA21. Method:The human lung cancer A549 cells were routinely passaged and divided into the blank group， blank serum group， QYSL-containing serum group， and siRNA group. The prepared QYSL-containing serum was used for intervention， with the optimal concentration and action time determined in previous studies. The protein and mRNA expression levels of miRNA21 and related molecules in its target phosphatase and tensin homolog deleted on chromosome 10 （PTEN）/phosphatidylinositol 3-kinase （PI3K） signaling pathway were detected by real-time polymerase chain reaction （Real-time PCR） and Western blot assay. Result:The comparison with the blank serum group revealed that the mRNA expression levels of miRNA21 in the QYSL-containing serum group and the siRNA group were decreased， while the PTEN mRNA expression in the QYSL-containing serum group was increased， showing significant differences （<italic>P</italic><0.01）. Compared with the blank serum， the QYSL-containing serum and siRNA significantly down-regulated PI3K and mammalian target of rapamycin （mTOR） mRNA expression （<italic>P</italic><0.01）， whereas the QYSL-containing serum did not change the mRNA expression of protein kinase B （Akt）. The protein expression levels of PTEN in the QYSL-containing serum group and the siRNA group were obviously elevated in contrast to that in the blank serum group （<italic>P</italic><0.05）. Meanwhile， the protein expression levels of phosphorylated Akt （p-Akt） and phosphorylated mTOR （p-mTOR） evidently declined in the QYSL-containing serum group （<italic>P</italic><0.05）， but there was no significant reduction in total Akt and mTOR protein expression. The PI3K protein expression was slightly down-regulated， with no statistical significance. Conclusion:QYSL inhibits the transcription of miRNA21， increases the expression of PTEN， and reduces the expression of key molecules in PI3K/Akt/mTOR signaling pathway， thus mildly inhibiting the "addiction" of lung cancer cells to oncogenes and blocking their proliferation.
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Objective:To explore the diagnostic value of serum miRNA-145 (miR-145), miRNA-21 (miR-21) and miRNA-197 (miR-197) for non-small cell lung cancer (NSCLC).Methods:The clinical data of 65 NSCLC patients in Luzhou Hospital of Traditional Chinese Medicine in Sichuan Province from January 2019 to June 2020 were retrospectively analyzed, and 60 healthy physical examiners were selected as the healthy control group. The relative expressions of serum miR-145, miR-21 and miR-197 in all subjects were detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR), and their relationship with clinicopathological features of NSCLC patients was analyzed. The receiver operating characteristic (ROC) curve was used to analyze the efficacy of miR-145, miR-21 and miR-197 alone and in combination in the diagnosis of NSCLC.Results:The relative expression of miR-145 in the NSCLC group was lower than that in the healthy control group (1.05±0.40 vs. 1.38±0.44, t = -4.326, P < 0.01), and the relative expressions of miR-21 and miR-197 were higher than those in the control group (2.37±0.89 vs. 0.83±0.25, 4.42±0.75 vs. 1.10±0.33, both P < 0.01). In the NSCLC group, the relative expressions of miR-21 and miR-197 in patients with TNM stage Ⅲ-Ⅳ were higher than those in patients with TNM stage Ⅰ-Ⅱ (both P < 0.01), and the relative expression of miR-145 in patients with adenocarcinoma was higher than that in patients with squamous cell carcinoma ( P < 0.01), and the relative expression of miR-21 in patients with lymph node metastasis was higher than that in patients without lymph node metastasis ( P < 0.01). When miR-145, miR-21 and miR-197 were used alone to diagnose NSCLC, the area under the curve (AUC) was 0.842 (95% CI 0.795-0.907), 0.868 (95% CI 0.812-0.938) and 0.857 (95% CI 0.801-0.913), and the best diagnostic cut-off values ??were 1.14, 2.03 and 2.98. When the best cut-off value was used for diagnosis, miR-145 alone was the most sensitive (83.76%), and miR-21 alone was the most specific (87.63%). The AUC for the combined detection of the three was 0.939 (95% CI 0.904-0.965), the diagnostic sensitivity was 91.58%, and the specificity was 79.86%. Conclusions:The relative expressions of miR-145, miR-21 and miR-197 are related to the pathological type, TNM staging and lymph node metastasis of NSCLC patients, and they may become markers for the auxiliary diagnosis of NSCLC. The combined detection of the three can improve the diagnostic efficiency and sensitivity.
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During the traumatic brain injury (TBI), improved expression of circulatory miR-21 serves as a diagnostic feature. Low levels of exosome-miR-21 in the brain can effectively improve neuroinflammation and blood-brain barrier (BBB) permeability, reduce nerve apoptosis, restore neural function and ameliorate TBI. We evaluated the role of macrophage derived exosomes-miR-21 (M-Exos-miR-21) in disrupting BBB, deteriorating TBI, and Rg1 interventions. IL-1
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@#Objective To explore the effect of expression of miRNA-21 on bone marrow mesenchymal stem cells (BMSCs). Methods In this study, flow cytometry was used to identify the surface-associated antigens of BMSCs. The 10 μmol/L 5-azacytidine was used to induce BMSCs to differentiate to cardiomyocyte-like cells. Immunofluorescence was used to detect the expression of troponin I (cTnI). The samples were assigned to 3 groups: a blank group, a miRNA-21 mimic group, and a negative control (NC) group. The proliferation of BMSCs was detected by methyl thiazolylte-trazolium (MTT), the apoptosis of BMSCs was analyzed by flow cytometry. Western-blotting was used to identify the expression of cTnI and myod in the BMSCs. Results The proliferation of BMSCs was increased, because of the over expression of miRNA-21. But the apoptotic rate of the BMSCs was slower in the miRNA-21 group, on account of the expression of miRNA-21 was higher than that in the NC group and the CK group. The expression of cTnI in the miRNA-21 group was higher than that in the NC group or the CK group. Conclusion The results suggest that the up-regulation of miRNA-21 enhances proliferation of BMSCs, reduces the apoptosis of BMSCs. miRNA-21 promotes the differentiation of BMSCs, which may pave the way for the treatment directed toward restoring miRNA-21 function for myocardial ischemia.
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Objective To study the correlation between serum miRNA-21 (miR-21) level and chemosensitivity and prognosis of osteosarcoma patients. Methods A total of 68 osteosarcoma patients who were treated in Shanxi Provincial Cancer Hospital from August 2016 to August 2017 were selected. All patients received routine chemotherapy after admission. They were followed up for one year. According to the effectiveness of chemotherapy, they were divided into effective group (52 cases) and ineffective group (16 cases). The general clinicopathological characteristics of the two groups were recorded. Univariate logistic regression model was used to analyze the serum miR-21 level and chemosensitivity in osteosarcoma patients. The prognostic efficacy of serum miR-21 was evaluated by receiver operating characteristic (ROC) curve. Results The serum miR-21 expression in the effective group was higher than that in the ineffective group (6.6 ±0.7 vs. 5.2 ±0.7), and the tumor metastasis rate in the effective group was lower than that in the ineffective group [33.9％ (18/52) vs. 37.5％ (6/16)], and the differences between the two groups were statistically significant (both P< 0.05). There were no significant differences in gender, age, body mass index (BMI), tumor location, size of tumors and TNM stage between the two groups (all P>0.05). Univariate logistic regression model showed that serum miR-21 and metastasis were the factors affecting chemosensitivity (both P<0.05). ROC curve showed that the area under the curve (AUC) of using serum miR-21 to predict the prognosis of osteosarcoma patients was 0.774. Youden index indicated that the best cut-off points of serum miR-21 and chemosensitivity for predicting the prognosis of osteosarcoma patients was 6.25. Conclusion Serum miR-21 level in osteosarcoma patients is significantly correlated with chemosensitivity and prognosis, and it can be used as an effective index to predict the effect of chemotherapy and prognosis of patients with osteosarcoma.
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Retinoblastoma (Rb) is one of the most common eye malignancies occur in childhood. The crucial roles of non-coding RNAs, particularly long non-coding RNAs (lncRNAs) and microRNAs (miRNAs), have been widely reported in Rb progression. In the present study, we found the expression of lncRNA T-cell leukemia/lymphoma 6 (TCL6) was significantly downregulated in Rb tissues and cell lines. Knockdown of lncRNA TCL6 promoted cell proliferation while reduced cell apoptosis in Rb cells. Moreover, lncRNA TCL6 serves as a sponge for miR-21, a previously-reported oncogenic miRNA in Rb, by direct targeting to negatively regulated miR-21 expression, therefore modulating Rb proliferation through miR-21. TCL6 overexpression inhibited Rb cell proliferation while miR-21 overexpression exerted an opposing effect; the effect of TCL6 overexpression was partially attenuated by miR-21 overexpression. PTEN/PI3K/AKT signaling pathway was involved in lncRNA TCL6/miR-21 axis modulating Rb cell proliferation. Taken together, lncRNA TCL6 serves as a tumor suppressor by acting as a sponge for miR-21 to counteract miR-21-mediated PTEN repression.
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Apoptosis , Cell Line , Cell Proliferation , MicroRNAs , Porifera , Repression, Psychology , Retinoblastoma , RNA, Long Noncoding , RNA, Untranslated , T-LymphocytesABSTRACT
Objective: To observe the influence of miRNA-21 in the radiosensitivity of the breast cancer cells, and to preliminarily explore its mechanism. Methods: The breast cancer T47D and MDA-MB-361 cells were irradiated with 0. 0. 2.5. and 5. 0 Gy y-ray. respectively; CCK8 assay was used to detect the survival rates of T47D and MDA-MB-361 cells and flow cytometry was used to analyze the cell cycle; real-time quantitative PCR was used to detect the expression levels of miRNA-21 in cells during 72 h. The T47D and MDA-MB-361 cells were transfected with anti-miRNA-21 sequence (anti-miRNA-21 group) and negative control sequence (negative control group)∗ respectively; the untransfected cells were set as blank control group. Real-time quantitative PCR was used to detect the expression levels of miRNA-21 in cells in various groups. After irradiation with 0. 0 and 5. 0 Gy y-ray. CCK8 assay was used to detect the survival rates, and flow cytometry was used to analyze the cell cycle. Results: After irradiation with 5. 0 Gy y-ray. the survival rate of T47D cells was significantly higher than that of the MDA-MB-361 cells (PC0.05). Compared with 0. 0 Gy radiation group, the percentages of T47D and MDA-MB-361 cells in G:
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Oncogenic microRNAs are essential components in regulating the gene expression of cancer cells. Especially miR21, which is a major player involved of tumor initiation, progression, invasion and metastasis in several cancers. The delivery of anti-miR21 sequences has significant potential for cancer treatment. Nevertheless, since anti-miR21 sequences are extremely unstable and they need to obtain certain concentration to function, it is intensely difficult to build an effective delivery system for them. The purpose of this work is to construct a self-assembled glutathione (GSH)-responsive system with tumor accumulation capacity for effective anti-miR21 delivery and cancer therapy. A novel drug delivery nanosphere carrying millions of anti-miR21 sequences was developed through the rolling circle transcription (RCT) method. GSH-responsive cationic polymer polyethyleneimine (pOEI) was synthesized to protect the nanosphere from degradation by Dicer or other RNase in normal cells and optimize the pompon-like nanoparticle to suitable size. Dehydroascorbic acid (DHA), a targeting molecule, which is a substrate of glucose transporter 1 (GLUT 1) and highly expressed on malignant tumor cells, was connected to pOEI through PEG, and then the polymer was used for contracting a RNA nanospheres into nanopompons. The anti-miR21 nanopompons showed its potential for effective cancer therapy.
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<p><b>OBJECTIVE</b>Arsenic is a metalloid environmental carcinogen involved in the occurrence and development of many cancers. miRNA-21 plays a crucial role in arsenic-induced carcinogenesis. We aimed to elucidate the mechanism by which miRNA-21 influences arsenic-induced cancer.</p><p><b>METHODS</b>We used meta-analysis of published studies to determine how arsenic induces cancerous cells through miRNA-21.</p><p><b>RESULTS</b>Low-dose arsenic exposure (⪕ 5 μmol/L) can increase miRNA-21 and phosphorylated signal transducter and activator of transcription 3 (pSTAT3) expression, and decrease programmed cell death protein 4 (PDCD4) and protein sprouty homolog 1 (Spry1) expression. High-dose arsenic exposure (> 5 μmol/L), can increase miRNA-21 expression, and decrease Spry1 and E-cadherin expression. Short-term arsenic exposure (⪕ 24 h) can increase miRNA-21 and pSTAT3 expression, and decrease PDCD4 expression. Moreover, long-term arsenic exposure (> 24 h) can increase the miRNA-21, STAT3, and pSTAT3 expression, and decrease PDCD4 expression. We found that activation of miRNA-21 and pSTAT3 were most pronounced following long-term arsenic exposure at low doses, and the effects on PDCD4 expression were most pronounced following short-term arsenic exposure at low doses. miRNA-21 inhibitors increased the expression of tumor suppressor genes PDCD4, PTEN, and Spry1 and miRNA-21-mimics suppressed the expression of these tumor suppressor genes.</p><p><b>CONCLUSION</b>Arsenic can cause cancer by activating miRNA-21 and inhibiting the expression of PDCD4, PTEN, and Spry1.</p>
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Objective To investigate the expression of miRNA (miR-21) in mucosa-associated lymphoid tissue(MALT) lymphomas and its relationship with the effects in CHOP regimen. Methods Lymph gland tissues and preoperative peripheral blood of 52 patients pathologically diagnosed with MALT lymphoma in Department of Hematology of Affiliated Hospital of Hebei University of Engineering from January 2015 to December 2017 were collected; meanwhile, 10 tissues from patients with lymphadenitis and 20 peripheral serum from healthy examination patients were also collected. Quantitative real-time polymerase chain reaction (qPCR) was used to compare the lymph gland tissues in MALT lymphoma patients and lymphadenitis patients, and the expression of miR-21 in peripheral serum of MALT lymphoma and healthy people. The selected 20 cases of MALT lymphoma were given CHOP regimen treatment for 6 cycles, and then the efficiency and inefficiency were compared; at the same time, the expression of miR-21 in peripheral serum of the patients in the effective group was detected. Results The relative expression of miR-21 of lymph node tissues in lymphadenitis patients and MALT lymphoma patients was 1.03±0.12 and 4.53±0.73 respectively, and the relative expression of miR-21 in preoperative peripheral serum of healthy people and MALT lymphoma patients was 0.83±0.04 and 3 . 87 ± 0 . 21 respectively , and the differences were statistically significant (P ＝ 0.047, P ＝ 0.044). After 6 cycles of CHOP regimen, the total effect rate of MALT lymphoma was 70 % (14/20), and the relative expression of miR-21 in preoperative and postoperative peripheral serum for the patients who obtained the effective results after CHOP regimen was 3.95 ±0.08 and 1.62 ±0.41, and the differences were statistically significant (P＝ 0.035). Conclusions The expression of miR-21 has a certain correlation with the occurrence and development of MALT lymphoma. Furthermore, the serum miR-21 expression may be related to the effect of CHOP regimen chemotherapy in MALT lymphoma.
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Objective To study the clinical application of serum miRNA 21 and miRNA-126 in diagnosis of the coronary artery restenosis after percutaneous coronary intervention (PCI).Methods The serum and clinical data of 82 cases of acute coronary syndrome (ACS) patients treated with PCI from the Hospital of Jingyang Country and the Affiliated Hospital of Shaanxi University of Chinese Traditional Medicine (stenosis group consisted of 30 cases,non-stenosis group consisted of 52 cases),and healthy persons (control group consisted of 33 cases) were collected.The relative expression levels of serum miRNA-21 and miRNA 126 were detected using real-time qPCR technology in the comparative analysis for their changes in the groups.Intravascular ultrasound (IVUS) was used to evaluate the occurrence of restenosis after PCI.Results Serum miRNA-21 and miRNA 126 levels in the control group,the non-stenosis group and the stenosis group were,,(0.22 ± 0.03,0.43±0.08,0.92±0.17) and (0.87±0.15,0.49±0.10,0.23±0.04),respectively.Compared with the control group,the expression level of serum miRNA-21 in the stenosis group and the non stenosis group were significantly increased,while the miRNA-126 levels was significantly decreased (P=0.000),the differences were statistically significant.Compared with the non stenosis group,the level of miRNA-21 in the stenosis group was significantly increased,while the level of miRNA-126 was significantly decreased (P=0.000),the differences were statistically significant.The results of IVUS showed that the plaque area (PLA) of the patients in the stenosis group was significantly higher than that in the non stenosis group,while the minimal luman area (MLA) was significantly decreased,and the differences were statistically significant (P=0.000).There was a negative correlation between the expression of two markers in the stenosis group (r=-0.919,P＜0.01).The levels of miRNA-126 and miRNA 21 in the stenosis group were correlated with PLA and MLA (r=-0.945,0.926,P＜ 0.01) and (r=0.933,-0.917,P＜0.01).Conclusion The detection of serum miRNA-21 and miRNA-126 levels can be used in diagnosis and prediction for the coronary artary restenosis of ACS after PCI,as the gene detection index of the disease.
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OBJECTIVE@#To observe the expression differences of the plasma miRNA-1, miRNA-21 between patients with coronary heart disease (CHD) and without coronary artery lesions, between patients with in-stent restenosis (ISR) and none in-stent restenosis (NISR), and to study their predictive value for ISR occurred after percutaneous coronary intervention (PCI) in patients with CHD and diabetes mellitus (DM).@*METHODS@#The selected subjects were divided into CHD group in which patients were implemented stenting (=187), and control group in which patients were without coronary artery lesions (=195). According to the guidelines, the control group was divided into normal group (=150), simple-DM group (=45); the CHD group was divided into simple-CHD group (=119) and CHD-DM group (=68), the CHD group was also divided into ISR group (=48), NISR group (=139), and the ISR group was divided into simple-ISR group (=26) and ISR-DM group (=22) again. Plasma was collected from each group, and total RNA was extracted, the level of blood miRNA-1, miRNA-21 of each group was detected, and their level differences were analyzed.@*RESULTS@#Compared with control group, the level of miRNA-1 and miRNA-21 of CHD group was increased (<0.05); compared with NISR group, the level of miRNA-1 and miRNA-21 of ISR group was increased (<0.05). The incidence of ISR of CHD-DM group was obviously higher than that of simple-CHD group, ISR-DM group's level of miRNA-21 was higher than that of simple-ISR group (<0.05), and there was no difference of miRNA-1 level between ISR and ISR-DM group (<0.05). In Logistics, for CHD patents, the OR of DM, miRNA-1, miRNA-21 were 2.132, 3.066, 1.924 respectively (<0.05); for CHD patents with ISR, the OR of DM, miRNA-21 were 2.123, 3.066 respectively (<0.05); especially for CHD and DM patents with ISR, the OR of miRNA-21 was 9.148 (<0.05). In ROC curve, for CHD patients with ISR, the AUC of miRNA-1, miRNA-21 were 0.854, 0.857 respectively; for CHD-DM patients with ISR, the AUC of miRNA-21 was 0.783.@*CONCLUSIONS@#To predict the occurrence of ISR for CHD patients, the plasma miRNA-1 and miRNA-21 have a relatively high specificity and sensitivity, for CHD patients with DM, miRNA-21 may have a higher clinical value.
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Humans , Coronary Angiography , Coronary Restenosis , General Surgery , Diabetes Complications , Diabetes Mellitus , MicroRNAs , Blood , Nerve Tissue Proteins , Blood , Percutaneous Coronary Intervention , StentsABSTRACT
To describe the relationship between miRNA-21 and renal fibrosis.Methods NRK-52E cells cultured in vitro,then constructed fibrosis model induced by transforming growth factorβ1 ( TGF-β1 ) , then bone morphogenetic protein-7 ( BMP-7 )-containing lentivirus vector was used to transfect the NRK-52E cells to anti-fibrosis, SMAD1, SMAD6, FN1, rno-miR-21-5p gene expression were detected.Results After TGF-β1 treatment, the BMP-7 mRNA and protein expressions were significantly decreased, FN1 mRNA and protein expressions were significantly increased ( P<0.05 ) . After transfected by BMP-7-containing lentivirus vector, BMP-7 mRNA expression significantly increased(P<0.05), FN1, SMAD6 and rno-miRNA-21-5p mRNA decreased, SMAD1 mRNA increased(P<0.05).Conclusion Epithelial-to-mesenchymal transition (EMT) is positively correlated with the miRNA21 and TGF-beta 1, its maybe related to the changes of BMP-7, SMAD1 and SMAD6.
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Objective:To explore the effects of radiation on miRNA-21 expression and the biology of human salivary gland adenoid cystic carcinoma cell lines(SACC-83,SACC-LM).Methods:In vitro cultured SACC-83 and SACC-LM cells were radiated by Varian 23 EX with the dose of 0(control),2,6 and 10 Gy respectively.Cell proliferation,apoptosis and miRNA-21 expression were observed by CCK-8 assay,flow cytometry and qRT-PCR methods respectively.Results:48 h after radiation the proliferation ability of SACC-LM cells decreased with the increasing dose of radiation(F =1 321.646,P =0.000),so did the miRNA-21 expression (F =177.964,P =0.011).However,the early apoptosis rate (F =354.484,P =0.039),the later apoptosis rate (F =254.278,P =0.042) and heteroploid ratio(F =1 562.991,P =0.001) increased with the increasing dose of radiation.In SACC-83 cell line 48 h after 6 Gy radiation the cell proliferation(F =1 537.214,P =0.013) and the miRNA-21 expression(F =134.868,P =0.017) were lower than that of other radiation doses.Moreover,the early apoptosis rate (F =88.579,P =0.006),the late apoptosis rate (F =1 391.345,P =0.033),heteroploid ratio(F =250.461,P =0.004) were higher than that of other radiation doses.Conclusion:The miRNA-21 expression in SACC-LM and SACC-83 cell lines is conversely associated with the radiation sensitivity.
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Objective To investigate the effect of sodium arsenite (NaAsO2) on the expression of miRNA-21 (miR-21) mediated by transcription factor ETS-1 in human normal hepatocytes (L-02).Methods Dose-effect study:The L-02 cells were treated with different doses of NaAsO2 [0.0 (control),2.5,5.0,10.0,20.0,40.0 μmol/L] for 24 h.Time-effect study:L-02 cells were exposed to 0 (control) and 20 μmol/L NaAsO2 for 12,24,36 and 48 h (n =6).ETS-1 and miR-21 were treated with ETS-1 shRNA and miR-21 inhibitor,respectively.The cells treated with ETS-1 shRNA (100 nmol/L) were divided into 4 groups:①ETS-1 shRNA NC treatment alone (control group);②ETS-1 shRNA NC combined with NaAsO2 (20 μ,mol/L) treatment group;③ETS-1 shRNA treatment alone group;④Treatment with ETS-1 shRNA and NaAsO2 (20 μmol/L) group.The MiR-21 inhibitor (100 nmol/L) treated cells were also divided into 4 groups:① miR-21 inhibitor NC treatment (control group);② miR-21 inhibitor NC combined with NaAsO2 (20 μmol/L);③miR-21 inhibitor group;④miR-21 inhibitor combined with NaAsO2 (20 μ mol/L) treatment group.The expression of ETS-1 mRNA and miR-21 were detected by quantitative real-time PCR (qRT-PCR);the protein expression of ETS-1 was detected by Western blotting.Results Dose-effect study:The expression of ETS-1 mRNA in the groups of 0.0 (control),2.5,5.0,10.0,20.0 and 40.0 μmol/L was 1.008 ± 0.028,1.552 ± 0.029,1.697 ± 0.050,1.842 ± 0.077,2.233 ± 0.096 and 2.235 ± 0.092;miR-21 expression was 1.025 ± 0.094,1.552 ± 0.072,1.683 ± 0.066,1.915 ± 0.171,2.337 ± 0.195 and 2.592 ± 0.177;the expression of ETS-1 protein was 1.060 ± 0.045,1.267 ± 0.160,1.386 ± 0.087,1.723 ± 0.196,2.208 ± 0.122 and 2.284 ± 0.224,respectively,and the differences were statistically significant (F =47.797,8.959,65.748,all P ＜ 0.05),the NaAsO2 dose groups were significantly higher than those of the control group (all P ＜ 0.05),and there was a dose-effect relationship.Time-effect study:The expression of ETS-1 mRNA in L-02 cells was 1.253 ± 0.175,1.623 ± 0.220,1.771 ± 0.324 and 1.913 ± 0.251,respectively at 12,24,36 and 48 h;the expression of miR-21 was 1.502 ± 0.111,1.716 ± 0.113,1.979 ± 0.186 and 2.452 ± 0.304;the expression of ETS-1 protein was 1.196 ± 0.105,1.502 ± 0.076,1.651 ± 0.074 and 1.839 ± 0.139,respectively,there were significant differences between the groups (F =14.936,39.180,39.441,all P ＜ 0.05).The expression of various time points of exposure to NaAsO2 was significantly higher than those in the control group (1.044 ± 0.115,1.044 ± 0.124,1.108 ± 0.088,1.053 ± 0.061;1.092 ± 0.061,1.096 ± 0.169,1.024 ± 0.111,1.057 ± 0.146;1.020 ± 0.017,1.049 ± 0.121,1.024 ± 0.089,1.031 ± 0.124,all P＜ 0.05),and there was a time-effect relationship.ETS-1 shRNA and miR-21 inhibitor treatment:compared with ETS-1 shRNA NC combined with NaAsO2 (20 μmol/L),ETS-1 shRNA and NaAsO2 (20 μmol/L) could significantly inhibit the expression of ETS-1 (0.912 ± 0.238 vs 1.641 ± 0.225,P ＜ 0.05),and down-regulated the expression of miR-21 (1.313 ± 0.334 vs 2.363 ± 0.252,P ＜ 0.05).There was no significant difference of ETS-1 mRNA expression between miR-21 inhibitor and NaAsO2 (20.μmol/L) group (1.580 ± 0.077 vs 1.576 ± 0.065,P ＞ 0.05) compared with miR-21 inhibitor NC and NaAsO2 (20 μmol/L).Conclusions The expression of ETS-1 and miR-21 in L-02 cells is significantly higher than those in control.The high expression of ETS-1 mediates NaAsO2-induced miR-21 overexpression,which may be an important molecular mechanism of arsenic-induced expression dysregulation of human hepatic miRNAs and liver damage.
ABSTRACT
Objective To explore the clinical significance of detection of plasma microRNA-21,-143 in identifying early esophageal cancer and esophageal non-tumor diseases.Methods The expression of plasma microRNA-21,-143 in 27 cases of patients with early esophageal cancer (esophagus cancer group),25 cases of patients with non-esophageal tumor (non-esophageal tumor group)and in the healthy controls were detected by RT-PCR,and detected the levels of plasma CEA and CA72-4 by the electrochemical luminescence technology,which of changes were analysed to observe the relationship between the changes and the esophageal cancer,the benign esophageal diseases for the two markers.Results The expression of plasma microRNA-21,-143 in the esophagus cancer group were 0.93±0.17,0.27±0.05,which of ones in the non-esophagus cancer group were 0.25±0.03,0.99±0.15,and with those in the control group were 0.23±0.03,1.02±0.15.Compared with those in the non-esophagus cancer group,the expression of plasma microRNA-21,-143 were obviously up or down-regulated with significant differences (t=10.87,11.55,P＜0.01).Compared with those in the control group,which of ones were obviously up or down-regulated with significant differences (t=9.20,9.07,P＜0.01),and with no statistical significances in comparison between the esophagus cancer group and the control group (t=1.39,1.19,P＞0.05).The positiverate of plasma microRNA-21,-143 in the esophageal cancer,non-esophagus cancer group and the control group were,81.4 ％ (22/27),4.0 ％(1/25) and 0 (0/24);85.1％ (23/27),4.0％ (1/25),and 0 (0/24),respectively.The positive rate of microRNA-21,-143 in the esophageal group respectively in comparison with those in the non-esophagus cancer group and the control group were significantly higher,the differences had statistical significances (x2 =31.59,34.39,P＜ 0.01;x2 =34.42,37.23,P＜ 0.01).The expression of two markers in the esophagus cancer group were no statistically significant differences compared with control group (x2 =0.980,0.980,P＞0.05).The sensitivity and specificity of microRNA-21,-143 in early diagnosis on the esophageal cancer were 81.4 ％,97.9 ％ and 85.1％,97.9 ％.The sensitivity of microRNA-21,-143 in the esophageal group were significantly higher compared with those of CEA and CA72-4,the differences were statistically significant (x2 =12.79,P＜0.01;x2 =5.33,P＜0.05;x2 =15.03,P＜0.01;x2 =6.95,P＜0.05).The specificity of microRNA-21,-143 in the esophageal cancer group were no statistically significant differences in comparison with those of CEA and CA72-4 (x2 =1.043,0.000,P＞0.05) and (x2=1.043,0.000,P＞0.05),respectively.The analysis results from the spearman correlation test showed that in the esophageal cancer group,the expression of plasma microRNA-21,-143 had a negative correlation (r =0.658,P＜0.01).Which of ones respectively associated with the levels of CEA and CA72-4 (r=0.607,0.623,P＜0.01 and r=0.579,0.610,P＜0.01).Conclution The detection of expression of plasma miRNA-21,miRNA-143 in the patients with the early esophageal cancer and non-esophageal tumor can provide a new train of thought for pathologic diagnosis of early esophageal cancer.