MicroRNA-150-5p inhibits the proliferation and invasion of human larynx epidermiod cancer cells though regulating peptidyl-prolyl cis/trans isomerase

Abstract Objective This study aimed to investigate the molecular mechanism of miR-150-5p regulating the malignant biological behavior of Human Epidermoid cancer cell (HEp-2) by targeting peptidyl-prolyl cis/trans isomerase NIMA-Interacting-1 (PIN1). Methods Firstly, qRT-PCR and Western blot were adopted to detect the expression levels of miR-150-5p and PIN1 in cancer tissue and paracancerous tissues of patients with LSCC, and those in human bronchial epithelial cells (16 HBE) and HEp-2. Next, the targeted relationship between miR-150-5p and PIN1 was assessed by bioinformatics website and dual-luciferase reporter assay, followed by their correlation analysis. Besides, after interfering with miR-150-5p or PIN1 expression in HEp-2 cells, CCK-8, cell colony formation assay, and transwell assay were utilized to detect the proliferation, viability, and invasion of cells, respectively. Subsequently, the protein levels of MMP-2, MMP-9, and EMT-related proteins in HEp-2 cells were checked by Western blot. Results Expression of miR-150-5p was down-regulated in LSCC tissues and HEp-2 cells. Moreover, miR-150-5p suppressed proliferation and invasion of HEp-2 cells, affected protein expression related to MMP and EMT, thereby inhibiting development of cancer. The expression of PIN1 was significantly increased in cancer tissues and HEp-2 cells, and there was a targeted relationship and negative correlation between miR-150-5p and PIN1 in cancer tissue. However, overexpression of PIN1 could reverse the effect of miR-150-5p on the proliferation and invasion of HEp-2 cells. Conclusion In a nutshell, there is a targeted relationship between PIN1 and miR-150-5p. Besides, miR-150-5p can inhibit the proliferation and invasion of HEp-2 cells by regulating the expression of PIN1. Level of evidence 3.

Cepharanthine promotes A549 cell apoptosis by regulating miR-150 and miR-182 / 中国病理生理杂志

AIM:To investigate the effect of cepharanthine on the growth of human lung carcinoma A549 cells and the underlying mechanism. METHODS:After A549 cells were treated with cepharanthine,the growth inhibitory rate was detected by MTT assay. The cell morphological changes were observed under light microscope. The apoptosis of the A549 cells was analyzed by flow cytometry. The expression of microRNA (miR)-150, miR-182, p53 mRNA and FOXO1 mRNA were detected by real-time PCR. The downstream target genes were predicted by software,and the expression of p53 and FOXO1 was determined by Western blot. RESULTS:After cepharanthine treatment,the growth of A549 cells was in-hibited,the apoptosis rate was significantly increased,and the expression levels of miR-150 and miR-182 were significantly decreased. With cepharanthine treatment at 10 μmol/L,the expression levels of p53 and FOXO1 were elevated;however, with cepharanthine at 30 μmol/L,the expression levels of p53 and FOXO1 were decreased. After transfection with miR-150,the expression of p53 was significantly decreased, while the expression of FOXO1 was significantly decreased after transfection with miR-182. CONCLUSION:Cepharanthine inhibits the growth of A549 cells and promotes the apoptosis of A549 cells by inhibiting the expression of miR-150 and miR-182. miR-150 and miR-182 may down-regulate the expression of p53 and FOXO1,respectively.

MiR-150 improves cardiac fibrosis after myocardial infarction by targeting c-Myb / 实用医学杂志

Objective To evaluate the role and mechanism of miR-150 in cardiac fibrosis after MI.Methods A rat model of MI was established by up-regulating miR-150 through overexpressing miR-150 lentivirus.Real-time PCR and Western blot were applied in detecting the expression of collagen 1 α 1 and α-SMA protein in infarction area border.Masson coloration was applied in measuring fibrosis.Cardiac fibroblasts were isolated and cultured.UTR was used to report the carrier and lentivirus.And c-Myb siRNA was used to verify the relationship between c-Myb and microRNA-150.Results In vivo,MiR-150 was down-regulated in myocardium border zone in 14 day and 28 day after infarction (P ＜ 0.001,P ＜ 0.05),and overexpressing miR-150 promoted myocardial fibrosis (P ＜ 0.001),and inhibited the expression of collagen1α 1 and α-SMA (P ＜ 0.01,P ＜ 0.05).In vitro,c-Myb was the direct target gene of miR-150,and inhibited the expression of c-Myb resulting in the down regulation of collagen1α 1 and α-SMA,suggesting that the role of miR-150 was achieved by regulating c-Myb.Conclusions MiR-150 was down-regulated in myocardium border zone,and myocardial fibrosis can be improved by targeting c-Myb.

Role of microRNA-150 in proliferation, migration and invasion of conjunctival mucosa-associated lymphoid tissue lymphoma / 第二军医大学学报

Objective To observe the expression of microRNA-150 (miR-150) in conjunctival mucosa-associated lymphoid tissue (MALT) lymphoma, so as to investigate its mechanism in proliferation, migration and invasion of conjunctival MALT lymphoma. Methods The expressions of ;m'P-150 and Cbl-b, a possible downstream molecule of miR-150, were measured by qPCR in MALT lymphoma tissues and precancerous tissues collected from 3 patients with conjunctival MALT lymphoma in Changzheng Hospital of Second Military Medical University. Then, we transfected miR-150 inhibitor and negative control into human multiple myeloma cell lines RPM1 8226 by cell transfection. CCK-8 assay and flow cytometry (FCM) method were used to investigate the role of miR-150 in the proliferation and apoptosis of RPM1 8226 cells. Transwell assay was used to analyze the effect of miR-150 on the migration and invasion of RPMI 8226 cells. Western blotting analysis was used to examine the regulation of miR150 on Cbl-b expression in RPMI 8226 cells. Results The expression of miR-150 was significantly increased in the conjunctival MALT lymphoma tissues compared with precancerous tissues (P< 0. 05,P< 0. 01). Compared with negative control group, the proliferation of RPMI 8226 cells was significantly repressed (P< 0. 01), the apoptosis was significantly increased (P< 0. 01), and the migration and invasion were significantly decreased (P< 0. 05,P< 0. 01) after transfection of miR-150 inhibitor. The expression of Cbl-b was significantly up-regulated in MALT lymphoma tissues, and was significantly increased after inhibiting miR-150 expression. Conclusion Up-regulated miR-150 can promote the proliferation, migration and invasion of lymphoma cells and is involved in the generation of conjunctival MALT lymphoma, which may be mediated by inhibiting its downstream target gene Cbl-b.

MicroRNA-150 regulates differentiation of NK and NKT cells through decre-asing CD122 expression / 中国免疫学杂志

To explore the effects of microRNA-150 deletion on the development and homeostasis of regulatory T cells (Treg),γδT cells,NK and NKT cells.Methods:microRNA-150 knockout mice were used and microRNA-150 expression was detected by Real-time PCR.The numbers of Treg ,γδT,NK and NKT cells in the thymus and spleen of normal control and microRNA-150 knockout mice were detected by Flow cytometry.Cell apoptosis was detected by Annexin V staining , and cell proliferation was detected by 5-Bromo-2-deoxyUridine ( Brdu ) incorporation.Results: microRNA-150 deletion did not affect the development and homeostasis of regulatory T cells (Treg) andγδT cells.However,microRNA-150 deletion resulted in a significant reduction of the NK and thymic NKT cell number.In addition, microRNA-150 deleted NK and NKT cells showed an arrested developmental and maturational phenotype with a reduced expression of NK 1.1 and CD122.Moreover , cell apoptosis was significantly increased in microRNA-150 deleted NK and thymic NKT cells ,while a lower cell proliferation rate was shown in the microRNA-150 deleted NK but not NKT cells.Conclusion: CD122 may play an important role in the development and homeostasis of mouse NK and NKT cells regulated by microRNA-150.