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Aim: To explore the effects of microRNA- 27a-3p(miR-27a-3p) on collagen type I (Col I) and collagen type III (Col III) synthesis in pulmonary fibroblasts and the underlying mechanisms. Methods: Human pulmonary fibroblasts MRC-5 were cultured and then transfected with miR-27a-3p mimic or its inhibitor. qPCR and Western blot were used to detect miR- 27a-3p levels and nuclear β-catenin content, respectively. The expression of Col I, Col III and Wnt3a was measured using qPCR and Western blot. Bioinformatics predicted the potential of miR-27a-3p bound to Wnt3a 3'-untranslated region (3'-UTR). Dual luciferase reporter gene analyzed the effects of miR-27a-3p mimic transfection on luciferase activity of wild type and mutant Wnt3a 3'-UTR. MRC-5 cells were treated with the Wnt3a/β-catenin signaling pathway inhibitor Dkkl, followed by transfection with miR-27a-3p mimic or its inhibitor. Col I and Col III expression was detected by qPCR and Western blot. Results: miR-27a- 3p mimic markedly increased miR-27a-3p levels and decreased Col I, Col III, Wnt3a and β-catenin expression (P 0. 05), revealing Wnt3a as a target of miR-27a-3p. In addition, Dkkl pretreatment almost fully reversed the inductive effect of miR-27a-3p inhibitor on Col I and Col III expression in MRC-5 cells (P < 0. 05). Conclusions: miR-27a-3p inhibits the biosynthesis of Col I and Col III in pulmonary fibroblasts, which is attributed to inactivated Wnt3a/β-catenin signaling pathway.
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Objective To investigate the miRNA expression profiles in rectal cancer tissues and their associations with clinical pathological stage, depth of tumor invasion, and lymph node metastasis, and to evaluate the potential of miRNA as diagnostic and prognostic markers of rectal cancer. Methods Human miRNA microarray was used to profile miRNA expression in rectal cancer tissues and matched adjacent normal tissues (n=71). The up-regulated miR-93-5p and down-regulated miR-27a-3p were screened out, and the top differentially expressed miRNA were validated by quantitative real-time polymerase chain reaction ( qRT-PCR) . The relationship between the expression of miRNA and clinical parameters was analyzed by ANOVA and Spearman correlation. Results The expression of miR-27a-3p was down-regulated in miRNA microarray, but was up-regulated in qRT-PCR analysis;the data were relatively discrete. The expression of miR-93-5p was up-regulated in both miRNA microarray and qRT-PCR analysis;the expression level of miR-93-5p in rectal cancer tissues was 3165 times that in adjacent normal tissues ( P=00058);the expression level was correlated with tumor volume ( P= 0004 ) , and was positively correlated with the level of carcinoembryonic antigen ( CEA) before treatment ( P=0001) and the number of lymph nodes metastases (rs=0534, P=0005). Conclusions There is a differential miRNA expression pattern between rectal cancer tissues and matched adjacent normal tissues. The miR-93-5p is highly up-regulated in rectal cancer tissues and may serve as a diagnostic and prognostic marker of rectal cancer.
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Objective To investigate the miRNA expression profiles in rectal cancer tissues and their associations with clinical pathological stage, depth of tumor invasion, and lymph node metastasis, and to evaluate the potential of miRNA as diagnostic and prognostic markers of rectal cancer. Methods Human miRNA microarray was used to profile miRNA expression in rectal cancer tissues and matched adjacent normal tissues (n=71). The up-regulated miR-93-5p and down-regulated miR-27a-3p were screened out, and the top differentially expressed miRNA were validated by quantitative real-time polymerase chain reaction ( qRT-PCR) . The relationship between the expression of miRNA and clinical parameters was analyzed by ANOVA and Spearman correlation. Results The expression of miR-27a-3p was down-regulated in miRNA microarray, but was up-regulated in qRT-PCR analysis;the data were relatively discrete. The expression of miR-93-5p was up-regulated in both miRNA microarray and qRT-PCR analysis;the expression level of miR-93-5p in rectal cancer tissues was 3165 times that in adjacent normal tissues ( P=00058);the expression level was correlated with tumor volume ( P= 0004 ) , and was positively correlated with the level of carcinoembryonic antigen ( CEA) before treatment ( P=0001) and the number of lymph nodes metastases (rs=0534, P=0005). Conclusions There is a differential miRNA expression pattern between rectal cancer tissues and matched adjacent normal tissues. The miR-93-5p is highly up-regulated in rectal cancer tissues and may serve as a diagnostic and prognostic marker of rectal cancer.
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OBJECTIVE:To investigate the expression difference and its mechanism of miR-27a-3p and HOXB8 protein in cer-vical cancer tissues of HPV16-positive and HPV16-negative patients. METHODS:A total of 120 patients with cervical cancer in our hospital during Jan. 2012-Jan. 2016 were divided into HPV16-positive group(60 cases)and HPV16-negative group(60 cases) according to HPV16 infection situation. The expression of miR-27a-3p mRNA and HOXB8 mRNA in cervical cancer tissue were de-tected by RT-qPCR. The expression of HOXB8 protein was detected by Western blotting assay. DNA methylation level of miR-27a-3p promoter region was detected by nested-down methylation specific PCR (nMS-PCR). Histone methylation level of miR-27a-3p promoter region was detected by chromatin immunoprecipitation PCR (CHIP-PCR). The expression of miR-27a-3p mRNA and HOXB8 mRNA were detected after transfecting miR-27a-3p mimic and inhibitor into Human cervical cancer cell line SiHa,respectively. RESULTS:The expression of miR-27a-3p in HPV16-positive group was significantly lower than HPV16-nega-tive group,while HOXB8 mRNA and protein expression,DNA and histone methylation levels of miR-27a-3p promoter region were significantly higher than HPV16-negative group,with statistical significance (P<0.05 or P<0.01). After transfecting miR-27a-3p mimic into SiHa cells,the expression of miR-27a-3p was increased significantly,while that of HOXB8 mRNA was de-creased significantly;after miR-27a-3p inhibitor transfection,the expression of miR-27a-3p was decreased significantly,while that of HOXB8 mRNA was increased significantly,with statistical significance(P<0.01). CONCLUSIONS:HPV16 may down-regu-late the expression of miR-27a-3p through DNA methylation and histone methylation of promoter region,so as to influence the gen-eration and development of cervical cancer. HOXB8 may be the target protein of miR-27a-3p.