Effects of miR-369-3p on the proliferation and apoptosis of bladder cancer cells by regulating the expression of VEGFC gene / 中国医师杂志

Objective To observe the effect of miR-369-3p on vascular endothelial growth factor C (VEGFC) gene in bladder cancer cells EJ and 5637 and observe its effect on cell proliferation and apoptosis.Methods Bladder cancer cell lines EJ and 5637 were transfected with miR-NC (control group) or transfected with miR-369-3p (experimental group).The target gene of miR-369-3p was predicted by Targets can target gene prediction software.Fluorescence real-time quantitative polymerase chain reaction (qPCR) was used to detect the changes of VEGFC at mRNA level.The changes of VEGFC,p-Raf-1 and phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2) at the protein level were detected by Western blotting.The effect of miR-369-3p on the proliferation of bladder cancer cells was detected by cell counting kit (CCK-8) and clone formation experiment.The effect of miR-369-3p on apoptosis was detected by flow cytometry.Results After transfection with miR-369-3p,the expression of VEGFC at mRNA and protein levels was significantly decreased (P ＜ 0.05),the expression of p-Raf-1 and p-ERK1/2 protein was significantly decreased,the cell proliferation ability was significantly decreased (P ＜ 0.05),the number of clones formed was significantly decreased (P ＜ 0.05) and the apoptosis was significantly increased (P ＜ 0.01).Conclusions miR-369-3p can inhibit the proliferation and promote the apoptosis of bladder cancer cells by interfering with the expression of VEGFC gene,which may provide a new target for the biological therapy of bladder cancer.

Effect of MicroRNA-9 on cell proliferation and collagen synthesis in hepatic stellate cells by down-regulation of TGFBR2 / 中国医师杂志

Objective To investigate the effect of MicroRNA-9 (miR-9) on cell proliferation and collagen synthesis in hepatic stellate cells (HSCs),and to explore the potential mechanism.Methods HSC-T6 cells were cultured and transfected with miR-9 mimics with lipofectamine 2000.After incubation 48 h,the cells were collected and total proteins and RNAs were extracted.The expression of miR-9 was detected by quantitative real time polymerase chain reaction (qRT-PCR).The protein expression of type Ⅰ collagen and type Ⅲ collagen were measured by Western blot.The methyl thiazolyl tetrazolium (MTT) method was used to asses the proliferation of HSC-T6 cells.The expression of transforming growth factor-β1 receptor 2 (TGFBR2) was detected by qRT-PCR and Western blot.Results Compared to the control group,miR-9 expression in HSCs was increased in the miR-9 mimics group (P ＜ 0.05),type Ⅰ and type Ⅲ collagen protein expression was reduced by (44 ± 2) ％ and (50 ± 3) ％ (P ＜ 0.01),respectively.The proliferation activity of HSCs was decreased by (48 ± 4)％ (P ＜ 0.05).The expression of TGFBR2 was inhibited in the miR-9 mimics group.Conclusions Upregulation of miR-9 plays a role on suppressing cell proliferation and collagen synthesis in HSCs.This process might be mediated by downregulation of TGFBR2.

Effect of miR-218 on proliferation and cell cycle of gastric cancer cells / 中国医师杂志

Objective To investigate the effect of miR-218 on cell proliferation and cell cycle in the gastric cancers (GC).Methods SGC-7901 and MGC-803 cells were transfected with miR-218 mimics.Meanwhile,SGC-7901 and MGC-803 cells were transfected with control mimics (Scramble mimics) as negative control.Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect the miR-218 expression in these transfected cells.The effect of miR-218 overexpression on cell proliferation was evaluated with direct cell counting and MTS [3-(4,5-dimethylthiazol-2-yl)-5 (3-carboxymethonyphenol)-2-(4-sulfophenyl)-2H-tetrazolium inner salt] assay.Moreover,the effect of miR-218 overexpression on cell cycle was examined by fluorescence activated cell sorting (FACS).Results miR-218 expression was remarkably induced in miR-218-transfected cells,miR-218 overexpression led to a significant decrease in cell proliferation rate compared to control cells (P ＜ 0.05).miR-218 overexpression induced GC G1 arrest.Conclusions miR-218 can suppress GC cell proliferation and block cell cycle progression.It maybe play an important role in tumorigenesis and development of GC.

The study of new targeted-delivery of micro RNAs to bone-metastatic prostate tumors / 中国医师杂志

Objective To construct a new gene delivery system based on atelocollagen (ATE),and explore that modified aptamer (APT),and APT-ATE/miRNA (miRNA-15a and miRNA-16-1) were successfully synthesized to treat bone-metastatic prostatic cancers.Methods Flow cytometry (FCM) analysis was used to characterize APT-ATE complex.The diameter and zeta potential of complexes were measured by Zetasizer Nano-ZS9.The prostatic cancer (PCa) distribution experiments were used to explore its biological characteristics and targeting ability of PCa cells (PC3 and LNCaP).The inhibition of APT-ATE complex on LNCaP cell was determined with the cholecystokinin (CCK)-8 assay.Results FCM results demonstrated the successful synthesis of ATE-APT complex.The cellular uptake of vectors was concentration-dependent.The gene expression in vitro indicated that the modification of APT could increase the efficiency of gene expression and PCa targeting ability of ATE vectors to LNCaP [prostate specific membrane antigen (PSMA) over-expressing prostate cancer cells].The result of biodistribution showed that the bone uptake of APT-ATE was higher than ATE-APT.Conclusions APT-ATE/miRNA might be useful for preclinical and clinical studies on the treatment of bone-metastatic PCa.

Let-7a is involved in the pathogenesis of chronic eosinophilic rhinosinusitis with nasal polyp by regulating BAFF / 中国医师杂志

Objective To explore the regulatory effect of miRNA on B-cell activating factor (BAFF) in chronic eosinophilic rhinosinusitis with nasal polyp (Eso CRSwNP).Methods Ten nasal mucosal samples were collected from Eso CRSwNP patients who were admitted and treated in our hospital between January 2012 and February 2013.Normal nasal mucosal tissues (n =10) served as control.The miRNA-targeting BAFF was predicted by bioinformatics tools.Immunohistochemistry and real time polymerase chain reaction (RT-PCR) were used to assess the protein expression of BAFF and the predicted miR-NA.The correlation between the predicted miRNA and BAFF was analyzed.Results Down-regulated expression of let-a was confirmed in Eos CRSwNP,while the BAFF protein expression was increased.Let-a was positively correlated with BAFF in nasal epithelia.Conclusions Let-a might contribute to mucosal eosinophilia in eosinophilic CRSwNP via targeting BAFF.

The effect of let-7b and miR-199a on B16F10 cell growth and proliferation / 中国医师杂志

Objective To confirm whether or not let-7b and miR-199a were significantly associated with malignant melanoma growth and proliferation. Methods An over -expression plasmid and an inhibitor, which targeted on let-7b and miR-199a, was constructed. B16F10 cells were divided into seven groups: control group, let-7b plasmid group, miR-199a plasmid group, empty plasmid group, let-7b inhibitor group, miR-199a inhibitor group, inhibitor control group. Foreign gene was transfected into B16F10 cells, let-7b and miR-199a expression were validated from RNA level, protein level and cell level. Results The relative let-7b or miR-199a gene expression of the let-7b plasmid group (3.8776±0.1372)and miR-199a plasmid group (2.8660±0.2821)were significantly higher than control group (P＜0.05), the relative let-7b or miR-199a gene expression of the let-7b inhibitor group (0.2057±0.0263) and miR-199a inhibitor group(0.2656±0.0253) were significantly lower than control group(P＜0.05). The cyclinD1 expression of the let-7b plasmid group(2.023±0.315) and let-7b inhibitor group (1.857±0.377) were significantly higher than control group (0.997±0.041) (P＜0.05), whereas, the Met expression of themiR-199a plasmid group (5.19±0.309) and miR-199a inhibitor group (4.87±0.044) were significantly higher than control group (2.2±0.198) (P＜0.05). The let-7b plasmid group and miR-199a plasmid group B16F10 cell growth rate were slower than control group, especially on the third day after transfection, the growth rate gradually dropped to the lowest value (P＜0.05). In addition, the apoptosis rates of the let-7b plasmid group and miR-199a plasmid group reach to (11.8±1.19)% and (11.3±1.59)%,which were significantly higher than control group (P＜0.05). Conclusions let-7b and miR-199a may be a negative regulator on the B16F10 cell growth and proliferation.