Yunyun Ma

MiR-429 up-regulation induces apoptosis and suppresses invasion by targeting Bcl-2 and SP-1 in esophageal carcinoma

PurposeMicroRNAs (miRNAs) may act as oncogenes or tumor suppressor genes and, as such, they may play a role in cancer development. We investigated miR-429 expression levels in a cohort of esophageal carcinomas (EC) to assess its impact on EC cell growth, apoptosis and invasion.MethodsqRT-PCR assays were used to quantify miR-429 expression levels in 32 paired EC samples and adjacent non-neoplastic tissues. Assays for cell growth, apoptosis, caspase activity and trans-well invasion were used to evaluate the effects of miR-429 expression on EC cells. Luciferase reporter and Western blotting assays were used to test whether the Bcl-2 and specificity protein 1 (SP1) mRNAs serve as major targets of miR-429.ResultsThe expression levels of miR-429 in EC tissues were found to be lower than those in adjacent non-neoplastic tissues (P < 0.05). This relatively low expression was found to be significantly associated with the occurrence of lymph node metastases (P < 0.05). Apoptosis and migration rates were found to be significantly higher in two EC-derived cell lines (EC9706 and KYSE30) transfected with a miR-429 agomir (P < 0.05). Subsequent Western blotting and luciferase reporter assays showed that miR-429 can bind to putative binding sites within the Bcl-2 and SP1 mRNA 3′ untranslated regions (UTRs) to reduce their expression.ConclusionsIn primary EC tissues miR-429 is expressed at low levels. Up-regulation of miR-429 inhibits invasion and promotes apoptosis in EC cells by targeting Bcl-2 and SP1. Our findings suggest that Bcl-2 and SP1 may serve as major targets of miR-429. This study paves the way for a better understanding of the mechanism underlying EC pathogenesis and the development of novel, targeted therapies.

miR-21 Down-Regulation Suppresses Cell Growth, Invasion and Induces Cell Apoptosis by Targeting FASL, TIMP3, and RECK Genes in Esophageal Carcinoma

BackgroundmiR-21 is overexpressed in esophageal squamous cell carcinoma (ESCC) and is thought to be correlated with the development of the cancer. The target gene of miR-21 including FASL, TIMP3 and RECK is revealed by researchers. miR-21 may be involved in the tumorgenesis of ESCC by targeting FASL, TIMP3 and RECK.AimsThe purpose of this study was to explore the mechanism of miR-21 in the development of ESCC.MethodsmiR-21 expression in ESCC and the matched non-malignant adjacent tissues (NMATs) was examined by qRT-PCR. Cell growth, cell apoptosis and cell invasion ability of EC9706 and EC-1 cells was examined after the cells were transfected with miR-21 inhibitor. The potential target genes of miR-21 including FASL, TIMP3 and RECK were examined by western blot and Luciferase reporter assay.ResultsmiR-21 expression was increased significantly in ESCC tissues compared with NMAT. miR-21 down-regulation inhibits cell growth, cell invasion and induces cells to apoptosis. FASL, TIMP3 and RECK are direct targets of miR-21.ConclusionsmiR-21 down-regulation inhibits cell growth, invasion and induces cells to apoptosis by targeting FASL, TIMP3 and RECK genes.

Effect of miR-335 upregulation on the apoptosis and invasion of lung cancer cell A549 and H1299.

MicroRNAs are small non-coding RNAs that may also function as oncogenes and tumor-suppressor genes, as the abnormal expression of microRNAs is associated with various human tumors. However, the effect of miR-335 on the lung cancer cells remains unclear. The aim of the paper was to study the expression of miR335 in non-small cell lung cancer (NSCLC) and miR335’s relation to the metastasis, invasion, and apoptosis in lung cancer cells A549 and H1299. qRT-PCR was used to identify the miR-335 expression. The effects of miR-335 on cell proliferation, apoptosis, and invasion were further analyzed. Luciferase reporter assay and Western blot were to verify Bcl-w and SP1 as potential major target genes of miR-335. Finally, the effect of Bcl-w on miR-335-induced cell survival was determined. Our results showed that miR-335 expression was significantly lower in NSCLC tissue, which was significantly associated with lymph node metastasis. In contrast to cells in blank and negative control groups, incidence of apoptosis was significantly higher (P < 0.05) and the number of cells migrating through matrigel was significantly lower (P < 0.05) in miR-335 mimics transfected group. Western blot and luciferase reporter assay demonstrated that miR-335 could bind to the putative binding sites in Bcl-w (or SP1) mRNA 3′-untranslated region to visibly lower the expression of Bcl-w (or SP1). The introduction of Bcl-w cDNA without 3′-untranslated region abrogated miR-335-induced cell survival. These results indicated that upregulation of miR-335 can simultaneously suppress the invasiveness and promote apoptosis of lung cancer cell A549 and H1299 by targeting Bcl-w and SP1. Therefore, miR-335 may be a potential therapeutic target in NSCLC treatment.

Isolation and characterization of ZZ1, a novel lytic phage that infects Acinetobacter baumannii clinical isolates.

BackgroundAcinetobacter baumannii, a significant nosocomial pathogen, has evolved resistance to almost all conventional antimicrobial drugs. Bacteriophage therapy is a potential alternative treatment for multidrug-resistant bacterial infections. In this study, one lytic bacteriophage, ZZ1, which infects A. baumannii and has a broad host range, was selected for characterization.ResultsPhage ZZ1 and 3 of its natural hosts, A. baumanni clinical isolates AB09V, AB0902, and AB0901, are described in this study. The 3 strains have different sensitivities to ZZ1, but they have the same sensitivity to antibiotics. They are resistant to almost all of the antibiotics tested, except for polymyxin. Several aspects of the life cycle of ZZ1 were investigated using the sensitive strain AB09V under optimal growth conditions. ZZ1 is highly infectious with a short latent period (9 min) and a large burst size (200 PFU/cell). It exhibited the most powerful antibacterial activity at temperatures ranging from 35°C to 39°C. Moreover, when ZZ1 alone was incubated at different pHs and different temperatures, the phage was stable over a wide pH range (4 to 9) and at extreme temperatures (between 50°C and 60°C). ZZ1 possesses a 100-nm icosahedral head containing double-stranded DNA with a total length of 166,682 bp and a 120-nm long contractile tail. Morphologically, it could be classified as a member of the Myoviridae family and the Caudovirales order. Bioinformatic analysis of the phage whole genome sequence further suggested that ZZ1 was more likely to be a new member of the Myoviridae phages. Most of the predicted ORFs of the phage were similar to the predicted ORFs from other Acinetobacter phages.ConclusionThe phage ZZ1 has a relatively broad lytic spectrum, high pH stability, strong heat resistance, and efficient antibacterial potential at body temperature. These characteristics greatly increase the utility of this phage as an antibacterial agent; thus, it should be further investigated.

Expression analysis of serum microRNAs in idiopathic pulmonary fibrosis

Idiopathic pulmonary fibrosis (IPF) is a disease of unknown etiology with considerable morbidity and mortality. Seeking informative diagnostic markers with greater clinical significance is essential for the early diagnosis of IPF. microRNAs (miRNAs or miRs) have emerged as novel serum diagnostic biomarkers for various diseases. In this study, we performed microarray analysis of the miRNA expression profile in the serum of patients with IPF compared to that of control subjects. We then performed a preliminary analysis of biological functions for the most differentially expressed miRNAs. Some of the microarray results were validated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The results from this study provide evidence to link the biological role of miRNAs to IPF, and suggest that miRNAs may undertake a variety of functions. Additionally, we found that the altered expression levels of miR-21, miR-155 and miR-101-3p were associated with forced vital capacity (FVC) and radiological features in IPF. Our data may serve as a basis for further investigation, preferably in large prospective studies, before miRNA can be used as a non-invasive screening tool for IPF in routine clinical practice.

Mir-655 up-regulation suppresses cell invasion by targeting pituitary tumor-transforming gene-1 in esophageal squamous cell carcinoma

BackgroundMicroRNAs (miRNAs) can act as either oncogenes or tumor suppressor genes under different conditions and thus can play a significant role in cancer development. We investigated miR-655 expression in a cohort of esophageal squamous cell carcinoma (ESCC) to assess the impact of this miRNA on ESCC cell invasion and metastasis.MethodsA qRT-PCR assay was used to quantify miR-655 expression levels in 34 paired ESCC samples and adjacent non-tumor tissues. Wound healing and transwell assays were used to evaluate the effects of miR-655 expression on the invasiveness of ESCC cells. Luciferase reporter and western blot assays were used to determine whether the mRNA encoding pituitary tumor-transforming gene-1 (PTTG1) is a major target of miR-655.ResultsThe expression level of miR-655 in ESCC tissues was found to be lower than in adjacent non-tumor tissues (P < 0.05). This relatively low expression level was significantly associated with the occurrence of lymph node metastases (P < 0.05). Migration rates were significantly lower for two ESCC-derived cell lines (EC9706 and KYSE150) transfected with miR-429 mimics (P < 0.05). Subsequent western blot and luciferase reporter assays demonstrated that miR-655 could bind to putative binding sites within the PTTG1 mRNA 3’-untranslated region (3’-UTR) and thus reduce the expression.ConclusionsmiR-655 is expressed at low levels in primary ESCC tissues, and up-regulation of miR-655 inhibits ESCC cell invasiveness by targeting PTTG1. Our findings suggest that PTTG1 may act as a major target of miR-655. This study improves our understanding of the mechanisms underlying ESCC pathogenesis and may promote the development of novel targeted therapies.

Myricetin exerts anti-proliferative, anti-invasive, and pro-apoptotic effects on esophageal carcinoma EC9706 and KYSE30 cells via RSK2

Myricetin, a common dietary flavonoid, is widely distributed in fruits and vegetables and is used as a health food supplement based on its anti-tumor properties. However, the effect and mechanisms of myricetin in esophageal carcinoma are not fully understood. Here, we demonstrated the effect of myricetin on the proliferation, apoptosis, and invasion of the esophageal carcinoma cell lines EC9706 and KYSE30 and explored the underlying mechanism and target protein(s) of myricetin. CCK-8 assay, transwell invasion assay, wound-healing assay, cell cycle analysis, and apoptosis assay were used to evaluate the effects of myricetin on cell proliferation, invasion, and apoptosis. Nude mouse tumor xenograft model was built to understand the interaction between myricetin and NTD RSK2. Pull-down assay was used to verify molecular mechanism. Myricetin inhibited proliferation and invasion and induced apoptosis of EC9706 and KYSE30 cells. Moreover, myricetin was shown to bind RSK2 through the NH2-terminal kinase domain. Finally, myricetin inhibited EC9706 and KYSE30 cell proliferation through Mad1 and induced cell apoptosis via Bad. Myricetin inhibits the proliferation and invasion and induces apoptosis in EC9706 and KYSE30 cells via RSK2. Myricetin exerts anti-proliferative, anti-invasive, and pro-apoptotic effects on esophageal carcinoma EC9706 and KYSE30 cells via RSK2. Our results provide novel insight into myricetin as a potential agent for the prevention and treatment of esophageal carcinoma.

Differential expression profiling of microRNAs and their potential involvement in esophageal squamous cell carcinoma

MicroRNAs are small, noncoding RNAs approximately 18–24 nucleotides in length that negatively regulate gene expression at the posttranscriptional and/or translational level by binding to complimentary sequences in the 3′-untranslated regions of target mRNAs. Growing evidence has indicated the important roles for different miRNA species in the development of different cancers. Therefore, miRNAs have the potential to become new biological markers for esophageal squamous cell carcinoma (ESCC) and to be applied in the diagnosis, prognosis, and targeted treatment of ESCC. In this study, we performed a miRNA microarray to analyze the miRNA expression profile in ESCC compared to normal tissues. Then, we made a preliminary analysis of the biological function for the most differentially expressed miRNAs and their potentially target genes regulated. Some microarray results were validated by performing quantitative RT-PCR. The study provided evidence that linked the biological role of miRNAs to ESCC and showed that miRNAs could undertake a variety of mechanisms. Additionally, we also found that altered miR-429 and miR-451 expression levels were associated with the occurrence of lymph node metastases and the differentiation status and TNM stage in ESCC. The study of miRNAs may lead to finding novel methods to diagnose, treat, and prevent ESCC.

Epidemiological Analysis of HTLV-1 and HTLV-2 Infection among Different Population in Central China

Background HTLV-1 and HTLV-2 are retroviruses linked etiologically to various human diseases, and both of them can be transmitted by vertical route, sexual intercourse, blood transfusion and intravenous drug use. Recently, some HTLV-infected cases have been reported and this virus is mainly present in the Southeast coastal areas in China, but has not been studied for the people in Central China. Objectives To know the epidemiologic patterns among different population samples in Central China and further identify risk factor for HTLV-1 and HTLV-2 infection. Methods From January 2008 to December 2011, 5480 blood samples were screened for HTLV-1/2 antibodies by using enzyme immunoassay, followed by Western Blot. Results The prevalence of HTLV-1 and HTLV-2 was found with infection rates 0.13% and 0.05% among all population samples for HTLV-1 and HTLV-2, respectively. The highest percentages of infection, 0.39% and 0.20%, were found in the high risk group, while only 0.06% and 0.03% in the blood donor group. There was only one case of HTLV-1 infection (0.11%) among patients with malignant hematological diseases. Of seven HTLV-1 positive cases, six were co-infected with HBV, two with HCV and one with HIV. Among three HTLV-2 positive individuals all were co-infected with HBV, one with HCV. Conclusions HTLV-1 and HTLV-2 have been detected in the Central China at low prevalence, with the higher infection rate among high risk group. It was also found that co-infection of HTLV-1/2 with HIV and HBV occurred, presumably due to their similar transmission routes. HTLV-1/2 antibody screen among certain population would be important to prevent the spread of the viruses.

Chloroquine inhibits MGC803 gastric cancer cell migration via the Toll-like receptor 9/nuclear factor kappa B signaling pathway

Stimulation of Toll‑like receptor 9 (TLR9) has been associated with invasion in various types of cancer cell in vitro. The present study aimed to evaluate the expression of TLR9 in MGC803 gastric cancer cells and investigate the effect of a non‑specific TLR9 inhibitor, chloroquine (CQ), on MGC803 cell migration via the TLR9/nuclear factor kappa B (NFκB) signaling pathway. The expression of TLR9 was investigated using reverse transcription polymerase chain reaction (RT‑PCR), flow cytometry and western blot analysis. The effects of CQ on MGC803 cell proliferation were measured by MTT colorimetric assay. The mRNA expression levels of cyclooxygenase‑2 (COX‑2), matrix metalloproteinase (MMP)‑2, MMP‑7 and NFκB p65 were evaluated by RT‑PCR in MGC803 cells stimulated by various concentrations of CQ. The migration of gastric cancer cells treated with CQ at 12, 24 and 36 h was measured by wound healing assay. The results indicated that MGC803 cells expressed TLR9 and that CQ had anti‑proliferative effects on MGC803 cells and inhibited mRNA expression of COX‑2, MMP‑2, MMP‑7 and NFκB p65 (P<0.05). Furthermore, CQ inhibited the bioactivity of NFκB p65 and prevented the migration of MGC803 cells in a dose‑dependent manner (P<0.05). In conclusion, the results indicated that the TLR9/NFκB signaling pathway was involved in gastric cancer cell migration and that CQ had anti‑tumor activity.