Abstract： Objective To observe the expression changes of microRNA 1 （miRNA-1） and microRNA 21（miRNA-21） after ischemic preconditioning （IPC）, ischemic postconditioning （IPO） and remote ischemic preconditioning （RIPC）in an ischemia-reperfusion rat heart model in vitro, as well as the expression of their target protein heat shock protein 70 （HSP70） and programmed cell death 4 （PDCD4）, and evaluate whether miRNA are involved in endogenous cardio-protective mechanism. Methods The Langendorff-perfused Sprague-Dawley rat hearts were randomly assigned into one of the four groups, control group （CON group, n=12）, ischemia preconditioning group （IPC group， n=12）?, ischemia postconditioning group （IPO group， n=12） and remote ischemia preconditioning group （RIPC group，n=12）. Cardiac function was digitalized and analyzed. The expression of HSP70, PDCD4, B-cell lymphoma/leukemia-2 （Bcl-2） and Bax was detected by Western blotting. The expression of miRNA-1 and miRNA-21 was detected by real-time reverse transcriotion-polymerase chain reaction （RT-PCR）. Assessment of cardiac infarct size and myocardial apoptosis was determined using triphenyltetrazolium chloride （TTC） assay and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay （TUNEL） assay respectively. Results The expressions of miRNA-1 and miRNA-21 were up-regulated in IPC group, but the expression of miRNA-1 was down-regulated in RIPC group and IPO group （P＜0.05）. The expressionsof PDCD4, HSP70 and Bax were down-regulated in ‘conditioning’ groups compared with CON group （P＜0.05）. The expression of Bcl-2 was not statistically different among the four groups. The infarct size and the myocardial apoptosis in ‘conditioning’ hearts were significantly decreased compared with CON group （P＜0.05）. Conclusion The expressions of the miRNA-1 and miRNA-21 are different in IPC, RIPC and IPO groups, and their target proteins are not inversely correlated with the miRNAs in all the ‘conditioning’ groups.
Objective To explore the microRNA （miRNA） expression changes and related miRNA characteristics of colorectal cancer （CRC） with hepatic metastasis by miRNA microarray. Methods The fresh specimens of primary CRC were collected in 10 patients during operation， which with hepatic metastasis or not. miRNA microarray analysis was performed to compare the miRNA expression levels in two groups. The different expression levels of miRNA were validated by quantitative real-time PCR analysis. Results A total of six dysregulated miRNAs were identified in the CRC patients with hepatic metastasis comparing with CRC patients without hepatic metastasis， including 3 up-regulated miRNAs （miR-224， miR-1236， and miR-622） and 3 down-regulated miRNAs （miR-155， miR-342-5p， and miR-363）， and the quantitative real-time PCR result of miR-224 consisted with the microarray finding. Conclusions miR-224 may be involved in the process of CRC with hepatic metastasis pathogenesis. miR-224 would be a research direction on a new biomarker or therapic method in CRC with hepatic metastasis.
Objective To summarize the domestic and abroad articles related to the research on the relation between microRNA （miRNA） and pancreatic cancer，and explore the important effects of miRNA expression patterns in diagnosis of pancreatic cancer. Methods “microRNA and pancreatic cancer” were searched as key words by PubMed and CNKI series full-text database retrieval systems from 2000 to 2012. Totally 60 English papers and 15 Chinese papers were obtained. Choice criteria：the basic research of miRNA and pancreatic cancer，the clinical research of miRNA and pancreatic cancer， and the prospect of miRNA in pancreatic cancer diagnosis and treatment. According to the choice criteria，31 papers were finally analyzed. Results The miRNA expression spectrum and specific miRNA expression such as miR-21，miR-34，miR-217，miR-196a，miR-10a，miR-155，miR-221，miR-222，miR-181a，miR-181b，miR-181d， and the family members of miR-200 and let-7 might be used as tumor markers to differentiate pancreatic cancer from normal pancreas，chronic pancreatitis or pancreatic endocrine tumors，and might be used as prognostic factor to predict the outcome. Conclusions miRNA expression spectrum are not only related to diagnosis of pancreatic cancer， but also have provided a new research direction and method for gene therapy of pancreatic cancer．
Objective To clarify the trends of expression levels of several up-regulated micro RNA (miRNA) in tissues of atrophic bone nonunion and mRNAs and proteins of their related target genes in osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs), and to explore their biological functions. Methods The hBMSCs were isolated from bone marrow of il iac bone by gradient centrifugation, and cultured. Osteogenic culture medium was used for osteogenic differentiation of the 4th generation of hBMSCs. The changes of corresponding miRNAs, mRNA and protein expression levels of related target genes were observed at 0 hour, 12 hours, 1 day, 2 days, 4 days, 7 days, and 14 days, by quantitative real-time PCR and Western blot. Results In the process of hBMSCs osteogenic differentiation, the mRNA and protein expression levels of osteoblastic target genes [alkal ine phosphatase l iver/bone/kidney (ALPL), bone morphogeneticprotein 2 (BMP-2), and platelet-derived factor alpha polypeptide (PDGF-A)] at most time points increased significantly whencompared with the values at 0 hour except that of BMP-2 decreased at 12 hours and 1 day, with maximum changes at 1 to 7 days. The miRNA expression levels, mRNA and protein expression levels changed significantly at different time points, while the trends of hsa-miRNA-149 and hsa-miRNA-654-5p changes were negatively correlated with the trends of ALPL and BMP-2 mRNA and protein expression changes respectively (P lt; 0.05). There was no obviously negative correlation between the trends of hsa-miRNA-221 change and PDGF-A change (P gt; 0.05). Conclusion In the osteogenic differentiation process of hBMSCs, hsa-miRNA-149 and hsa-miRNA-654-5p are closely related with the mRNA and protein regulation of ALPL and BMP-2, respectively.
Objective To investigate the effects of nucleus localization signal linked nucleic kinase substrate short peptide (NNS) conjugated chitosan (CS) (NNSCS) mediated the transfection of microRNA-140 (miR-140) in rabbit articular chondrocytes in vitro. Methods Recombinant plasmid GV268-miR-140 and empty plasmid GV268 were combined with NNSCS to form NNSCS/pDNA complexes, respectively. Chondrocytes were isolated and cultured through trypsin and collagenase digestion from articular cartilage of newborn New Zealand white rabbits. The second generation chondrocytes were divided into 3 intervention groups: normal cell control group (group A), NNSCS/GV268 empty plasmid transfection group (group B), and NNSCS/GV268-miR-140 transfection group (group C). NNSCS/GV268 and NNSCS/GV268-miR- 140 complexes were transiently transfected into cells of groups B and C. After transfection, real-time fluorescent quantitative PCR (RT-qPCR) was used to detect the expressions of exogenous miR-140; Annexin Ⅴ-FITC/PI double staining and MTT assay were used to detect the effect of exogenous miR-140 on apoptosis and proliferation of transfected chondrocytes; the expressions of Sox9, Aggrecan, and histone deacetylase 4 (Hdac4) were detected by RT-qPCR. Results RT-qPCR showed that the expression of miR-140 in group C was significantly higher than that in groups A and B (P<0.05). Compared with groups A and B, the apoptosis rate in group C was decreased and the proliferation activity was improved, Sox9 and Aggrecan gene expressions were significantly up-regulated, and Hdac4 gene expression was significantly down-regulated (P<0.05). There was no significant difference in above indexes between groups A and B (P>0.05). Conclusion Exogenous gene can be carried into the chondrocytes by NNSCS and expressed efficiently, the high expression of miR-140 can improve the biological activity of chondrocytes cultured in vitro, which provides important experimental basis for the treatment of cartilage damage diseases.
ObjectiveTo detect expressions of transient receptor potential channel C5 (TRPC5) and microRNA-320a (miR-320a) in thyroid cancer and explore clinical significances of them in thyroid cancer.MethodsThe expressions of TRPC5 and miR-320a mRNA in the thyroid cancer were investigated by searching the Ualcan database. While the expressions of TRPC5 and miR-320a mRNA in 80 cases of thyroid cancer, 35 cases of thyroid adenoma and 32 cases of normal thyroid tissues adjacent to thyroid adenoma tissues in the Zhengzhou Seventh People’s Hospital from March 2014 to March 2015 were tested. Real time PCR was used to detect the expressions of TRPC5 mRNA and miR-320a mRNA in the various tissues and Western blot was used to detect the TRPC5 protein in the thyroid cancer tissues. Therelationships between the expressions of TRPC5 and miR-320a mRNAs and clinicopathologic features of thyroid cancer were analyzed. The correlation between expressions of TRPC5 and miR-320a mRNA was analyzed by Pearson method. The risk factors influencing the prognosis were analyzed by univariate and multivariate Cox proportional hazards regression model.ResultsThe results of Ualcan database showed that the expression level of TRPC5 mRNA in the thyroid cancer was higher than that in the normal thyroid tissue (P<0.001), while the expression level of miR-320a mRNA was lower than that in the normal thyroid tissue (P<0.001). The results of clinical cases showed that the expression level of TRPC5 mRNA was significantly higher, while the expression of miR-320a mRNA was significantly lower in the thyroid cancer tissues as compared with the normal thyroid tissues (P<0.05). There was a negative correlation between the expression level of TRPC5 and miR-320a mRNA in the thyroid cancer (r=?0.653, P<0.001). The expressions of TRPC5 and miR-320a mRNA were correlated with the degree of differentiation, lymph node metastasis, and TNM stage (P<0.05). Kaplan-Meier survival curve analysis found that the patients with higher expression level of TRPC5 and lower expression level of miR-320a showed the poor prognosis, and multivariate analysis found that the lower tumor differentiation, later TNM stage, with lymph node metastasis, higher expression level of TRPC5 mRNA, and lower expression level of miR-320a mRNA were the risk factors affecting prognostic survival (P<0.05).ConclusionsFrom the database and clinical case data, it is concluded that TRPC5 mRNA is highly expressed, while miR-320a mRNA is lowly expressed in thyroid cancer tissues, and expressions of TRPC5 and miR-320a mRNA are related to degree of tumor differentiation, lymph node metastasis, TNM staging, and prognosis in patients with thyroid cancer. TRPC5 and miR-320a mRNA might be used as potential indicators for clinical and prognostic monitoring.
ObjectiveTo review the role of microRNA (miRNA) in skin development and wound healing. MethodsThe recent literature about miRNA in skin development and wound healing was reviewed and analyzed. ResultsmiRNA extensively involved in the development of the skin,including epidermal cell proliferation,differentiation,aging and hair follicle development;miR-203 known as the "skin-specific miRNA" can directly inhibit the expression of p63 and promote the differentiation of the epidermis.Meanwhile,miRNA also involved in various stages of skin regeneration and wound healing.Abnormal expression of miRNA is closely related with abnormal wound healing. ConclusionmiRNA play an important role in maintaining normal skin physiology and skin regeneration.To explore their roles in the healing of skin wounds and their regulatory mechanism can provide a new target for the treatment,which has a potential value and broad prospects.
Objective To search for the key microRNAs (miRNAs) involved in myocardial fibrosis in hypertrophic cardiomyopathy, and to further explore the mechanisms involved in the regulation of myocardial fibrosis. MethodsForty-two patients with hypertrophic cardiomyopathy diagnosed and treated surgically in West China Hospital of Sichuan University from January 2014 to June 2018 were selected, including 29 males and 13 females, with a median age of 46 (15-69) years. In the myocardial tissue of patients with hypertrophic cardiomyopathy with different degrees of fibrosis, miRNAs with significantly different expression were screened and further verified at the cellular level. By regulating the expression of the target miRNAs, the expressions of fibrosis-related proteins and target genes were detected respectively. Finally, the target-binding relationship was verified by dual-luciferase reporter gene detection. ResultsmiR-484 was up-regulated in severely fibrotic myocardial tissue and activated cardiac fibroblasts. After cardiac fibroblasts were activated by TGF-β1, the expression of miR-484 was significantly up-regulated, the expression of fibrosis-related proteins (CollagenⅠ, α-SMA) increased, and the expression of the target gene HIPK1 decreased (P<0.05). After inhibiting the expression of miR-484 by transfection of miR-484 antagomir, the expression of fibrosis-related proteins decreased, while expression of HIPK1 was up-regulated (P<0.05). The detection of dual luciferase reporter gene showed that the luciferase activity of the transfected WT-miRNA-484 mimics group was lower than that of the control group (P<0.05). ConclusionmiR-484 is a pro-fibrotic miRNA that participates in the process of myocardial fibrosis by negatively regulating the expression of HIPK1. It can be used as a regulatory target to provide a therapeutic strategy for myocardial fibrosis.
Objective To systematically evaluate the correlation between the expression of microRNA (miRNA)-21 and the prognosis of esophageal cancer. Methods PubMed, Cochrane Library, Embase, Wanfang Data, China National Knowledge Infrastructure and VIP Databases were searched by for the literature on the correlation between miRNA-21 and the prognosis of esophageal cancer till July 10, 2022. Two researchers independently performed literature screening, quality evaluation, and data extraction. Statistical analysis was conducted with Stata 14.0. Results A total of 13 articles were included, including 1 204 patients. The results of meta-analysis showed that: the overall survival (OS) of patients with high expression of miRNA-21 was lower than that of patients with low expression of miRNA-21 [hazard ratio (HR)=2.11, 95% confidence interval (CI) (1.56, 2.84), P<0.001]. miRNA-21 expression was not associated with disease free survival [HR=2.53, 95%CI (0.67, 8.22), P=0.182]. The OS of Asian patients with high expression of miRNA-21 was significantly lower [HR=2.44, 95%CI (1.71, 3.49), P=0.005], while the OS of non-Asian patients was not related to miRNA-21 expression [HR=1.34, 95%CI (0.94, 1.91), P=0.363]. The high expression of miRNA-21 was correlated with the decreased OS in patients with esophageal squamous cell carcinoma [HR=2.22, 95%CI (1.52, 3.26), P=0.001], while the OS in patients with esophageal adenocarcinoma was not correlated with the expression of miRNA-21 [HR=1.39, 95%CI (0.63, 3.06), P=0.409]. Conclusion The overexpression of miRNA-21 is associated with poor prognosis and might be regarded as a potential prognostic biomarker for patients with esophageal cancer.