Jifang Wen

Urinary microRNA-30a-5p is a potential biomarker for ovarian serous adenocarcinoma

MicroRNAs (miRNAs) can serve as biomarkers in human cancer. To determine the clinical value of urinary miRNAs for ovarian serous adenocarcinoma, we collected urine samples from 39 ovarian serous adenocarcinoma patients, 26 patients with benign gynecological disease and 30 healthy controls. The miRNA microarray data showed that only miR-30a-5p was upregulated and 37 miRNAs were downregulated in the urine samples of ovarian serous adenocarcinoma patients, when compared to healthy controls, which was confirmed after conducting quantitative PCR. The upregulation of urinary miR-30a-5p was closely associated with early stage of ovarian serous adenocarcinoma as well as lymphatic metastasis. Receiver operator characteristic (ROC) analysis demonstrated the potential use of urinary miR-30a-5p as a diagnostic marker for ovarian serous adenocarcinoma. Furthermore, a lower urine level of miR-30a-5p was found in 20 gastric cancer and 20 colon carcinoma patients when compared to ovarian serous adenocarcinoma, suggesting that the upregulation of urinary miR-30a-5p may be specific for ovarian serous adenocarcinoma. miR-30a-5p was also upregulated in ovarian serous adenocarcinoma tissues and cell lines, while urinary miR-30a-5p from ovarian cancer patients was notably reduced following the surgical removal of ovarian serous adenocarcinoma, suggesting that urinary miR-30a-5p was derived from the ovarian serous adenocarcinoma tissue. Notably, miR-30a-5p was concentrated with exosomes from the ovarian cancer cell supernatant or urine from ovarian serous adenocarcinoma patients, supporting a pathway for excretion into the urine. The results also showed that the knockdown of miR-30a-5p significantly inhibited the proliferation and migration of ovarian cancer cells. In summary, to the best of our knowledge, the present study provided the first evidence of increased miR-30a-5p in the urine of ovarian serous adeno-carcinoma patients, while the inhibition of miR-30a-5p suppressed the malignant phenotypes of ovarian cancer in vitro. Therefore, miR-30a-5p serves as a promising diagnostic and therapeutic target for ovarian serous adenocarcinoma.

Down-Regulated miRNA-214 Induces a Cell Cycle G1 Arrest in Gastric Cancer Cells by Up-Regulating the PTEN Protein

To detect the expression of miRNA-214 in human gastric cancer cell lines of BGC823, MKN45 and SGC7901, and to identify the effect of miRNA-214 on cell cycle and apoptosis of these cells. Expression of miRNA-214 in human normal gastric mucosal cell line GES-1 and human gastric cancer cell lines was detected by real-time reverse-transcription polymerase chain reaction. Antisense-miRNA-214 oligonucleotides were transfected transiently into gastric cancer cell lines to down-regulate the expression of miRNA-214. The cell cycle and apoptosis were studied by flow cytometry assay. PTEN, one of the target genes of miRNA-214 was detected by using of immunocytochemistry and Western blotting. MiRNA-214 was overexpressed in gastric cancer cell lines of BGC823, MKN45 and SGC7901 compared with normal gastric mucosal cell line GES-1. Antisense-miRNA-214 oligonucleotides significantly down-regulated the expression of miRNA-214, and increased the portion of G1-phase and decreased the portion of S-phase in BGC823 and MKN45 cells. The immunocytochemistry test and Western blotting analysis showed that the down-regulation of miRNA-214 could significantly up-regulate the expression of PTEN in BGC823 and MKN45 cells. MiRNA-214 is overexpressed in human gastric cancer cell lines of BGC823, MKN45 and SGC7901. The down-regulation of miRNA-214 could induce a G1 cell cycle arrest in them, the up-regulation of PTEN maybe one of the mechanism.

Expression and functions of the repressor element 1 (RE-1)-silencing transcription factor (REST) in breast cancer

The repressor element 1 (RE‐1)‐silencing transcription factor (REST), also known as the neuron‐restrictive silencer factor (NRSF) or repressor binding to the X2 box (XBR), REST/NRSF/XBR, is originally discovered as a transcriptional repressor of a large number of primarily terminal neuronal differentiation genes in non‐neuronal cells and neural stem cells (NSCs). Recently, the tumor‐suppressor function of REST is finally proved. However, the expression profile and function of REST in breast cancer are not very clear. In this study, the expression of REST was detected in breast cancer tissue by immunohistochemistry. The results showed that REST expression was significantly lower in breast cancer samples compared to normal and benign breast samples (P < 0.05). Furthermore, the shRNA approach was used to investigate the function of REST in human breast cancer cells. Knocking down REST expression by shRNA in the human breast cancer MCF‐7 cells resulted in an increase in cell proliferation, suppression in apoptosis, and reduced sensitivity to anticancer drug with a concurrent significantly up‐regulated expression of Bcl‐2. These data implied a significant role of REST in breast cancer. The reduced expression of REST might contribute to the breast cancer pathogenesis. J. Cell. Biochem. 110: 968–974, 2010.

Elevated expression of semaphorin 5A in human gastric cancer and its implication in carcinogenesis

AIMS Semaphorin 5A, a member of semaphorin family, was originally identified as axonal guidance factor functioning during neuronal development. Here, we investigated semaphorin 5A expression in gastric cancer and explored its roles in gastric carcinogenesis. MAIN METHODS The expression of semaphorin 5A was examined by reverse transcription-polymerase chain reaction (RT-PCR) analysis in six gastric cancer cell lines and detected by real-time RT-PCR and Western blotting in 30 pairs of primary gastric cancer and normal gastric mucosa tissues. RNA interference (RNAi) technique was used to generate a semaphorin 5A-silenced stable cell line. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and anchorage-independent growth as well as flow cytometry assays were conducted to examine the effect of siRNA-semaphorin 5A on gastric cancer cell growth, proliferation and apoptosis. KEY FINDINGS Semaphorin 5A was expressed in all human gastric cancer lines and the expression of semaphorin 5A was significantly higher in cancer tissues than that in normal mucosa tissues. siRNA-mediated semaphorin 5A knockdown significantly suppressed the proliferation and anchorage-independent growth, and induced the apoptosis of gastric cancer cell line SGC7901. SIGNIFICANCE The present study suggests that overexpression of semaphorin 5A may contribute to gastric carcinogenesis, which reveals a novel expression and function of semaphorin 5A outside the nervous system and adds more weight to our knowledge of semaphorin 5A.

Epidermal growth factor-like domain-containing protein 7 (EGFL7) enhances EGF receptor-AKT signaling, epithelial-mesenchymal transition, and metastasis of gastric cancer cells.

Epidermal growth factor-like domain-containing protein 7 (EGFL7) is upregulated in human epithelial tumors and so is a potential biomarker for malignancy. Indeed, previous studies have shown that high EGFL7 expression promotes infiltration and metastasis of gastric carcinoma. The epithelial–mesenchymal transition (EMT) initiates the metastatic cascade and endows cancer cells with invasive and migratory capacity; however, it is not known if EGFL7 promotes metastasis by triggering EMT. We found that EGFL7 was overexpressed in multiple human gastric cancer (GC) cell lines and that overexpression promoted cell invasion and migration as revealed by scratch wound and transwell migration assays. Conversely, shRNA-mediated EGFL7 knockdown reduced invasion and migration. Furthermore, EGFL7-overexpressing cells grew into larger tumors and were more likely to metastasize to the liver compared to underexpressing CG cells following subcutaneous injection in mice. EGFL7 overexpression protected GC cell lines against anoikis, providing a plausible mechanism for this enhanced metastatic capacity. In excised human gastric tumors, expression of EGFL7 was positively correlated with expression levels of the mesenchymal marker vimentin and the EMT-associated transcription repressor Snail, and negatively correlated with expression of the epithelial cell marker E-cadherin. In GC cell lines, EGFL7 knockdown reversed morphological signs of EMT and decreased both vimentin and Snail expression. In addition, EGFL7 overexpression promoted EGF receptor (EGFR) and protein kinase B (AKT) phospho-activation, effects markedly suppressed by the EGFR tyrosine kinase inhibitor AG1478. Moreover, AG1478 also reduced the elevated invasive and migratory capacity of GC cell lines overexpressing EGFL7. Collectively, these results strongly suggest that EGFL7 promotes metastasis by activating EMT through an EGFR−AKT−Snail signaling pathway. Disruption of EGFL7−EGFR−AKT−Snail signaling may a promising therapeutic strategy for gastric cancer.

Molecular characterization and expression pattern of an odorant receptor from the myiasis-causing blowfly, Lucilia sericata (Diptera: Calliphoridae)

The blowfly Lucilia sericata (Diptera: Calliphoridae) is a facultative ectoparasite that causes myiasis in both man and animals, leading to serious human health problems and economic losses in the livestock industry. Like other insects, olfaction of this species plays an important role in host location and is presumably mediated by a seven transmembrane receptor family. Here, we isolate and characterize LserOR1, which is the first candidate member of the odorant receptor gene family from L. sericata. LserOR1 displayed high amino acid conservation with previously identified Or83b orthologs from different insect species. The transcripts of LserOR1 were detected in the major olfactory organs including the antennae and maxillary palps, as well as in traditionally non-olfactory tissues such as the legs and female ovipositors. In developmental studies, a quantitative real-time PCR assay was developed and validated for determining the relative expression levels of LserOR1 during several stages. In contrast to its extremely high expression in the adult stage, LserOR1 expression was at the lowest level during the egg stage, and then increased to a peak through the first two larval stages before declining in the third-instar stage. These results suggest that a broadly expressed LserOR1 receptor is likely to be essential for olfactory sensory processes throughout the lifetime of L. sericata. The present study provides the information that may aid in the development of novel blowfly repellents using olfactory proteins as molecular targets.

Increased Expression of Prohibitin and its Relationship with Poor Prognosis in Esophageal Squamous Cell Carcinoma

Prohibitin, a potential tumor suppressor, has been shown to be an anti- proliferative protein, a regulator of cell-cycle progression and in apoptosis. Recently, it was found to be over-expressed in breast cancer and gastric cancer, and it has been suggested as a biomarker in those diseases. To clarify the role and the prognostic significance of prohibitin expression in esophageal squamous cell carcinoma (ESCC), we analyzed the expression in ESCC and their corresponding nonneoplastic epithelia tissues by immunohistochemistry(IHC), Western blotting and real-time quantitative reverse transcription polymerase chain reaction(QRT-PCR).The relationship between prohibitin expression and clinicopathological variables was examined by statistical analysis. The findings suggested the up-regulation of prohibitin play an important role in the carcinogenesis of ESCC. The over-expression of prihibitin was significantly correlated with the depth of tumor, lymph node metastasis, distant metastasis, lymphatic invasion and vascular invasion of ESCC. These results suggested that prohibitin(+), lymph node metastasis and distant metastasis could be the independent risk factors for worse prognosis in ESCC patients.

Down-regulation of Notch receptor signaling pathway induces caspase-dependent and caspase-independent apoptosis in lung squamous cell carcinoma cells.

Cao H, Hu Y, Wang P, Zhou J, Deng Z, Wen J. Down‐regulation of Notch receptor signaling pathway induces caspase‐dependent and caspase‐independent apoptosis in lung squamous cell carcinoma cells. APMIS 2012; 120: 441–50.

Knockdown of fascin1 expression suppresses the proliferation and metastasis of gastric cancer cells.

Aims: Our current investigation attempts to study the role of the fascin1 gene in growth and metastasis of gastric cancer cell line MKN45. Methods: Small interfering RNA (siRNA) was used to inhibit fascin1 expression in the human gastric cancer cell line MKN45. Expression of fascin1 in fascin1 siRNA transfected cells (sifascin1), non‐transfected cells (NT) and non‐specific fascin1 siRNA cells (CON) were examined by Western blotting and reverse transcription polymerase chain reaction (RT‐PCR). Cell growth ability in vitro was evaluated by MTT and clone formation assays. Cell mobility in vitro was examined by the Boyden chamber assay. Nude mice metastasis models were established by abdominal cavity transfer method. Tumour growth was evaluated by immunohistochemistry with proliferating cell nuclear antigen (PCNA). Results: Knockdown of fascin1 expression in MKN45 cells resulted in decreased cellular proliferative and migratory abilities. In vitro, the cloning efficiency of siFascin1 cells (34.2%) was significantly lower compared to that in NT (78.5%) (p < 0.05). The migration rate in siFascin1 cells was significantly decreased (33.7%) compared with NT cells (89.4%) (p < 0.05). In vivo, the cell proliferation rate was lower in siFascin1 cells (25.8%) compared to that in NT (75.0%) (p < 0.05). The number of tumour clones in the liver was significantly lower in siFascin1 cells (2.0 ± 1.1) compared to that in NT (5.1 ± 1.6) (p < 0.05). Conclusions: Our study demonstrates that down‐regulation of fascin1 suppresses the proliferation and migration of gastric cancer cells MKN45, suggesting that fascin1 siRNA may offer a novel potential gene therapy approach for human gastric cancer with fascin1 over‐expression.

Follicular dendritic cell sarcoma: a report of six cases and a review of the Chinese literature

GoalsThe main purpose of this study is to broaden the clinicopathological spectrum and increase recognition of follicular dendritic cell sarcoma (FDCS) through analysis of the clinical and pathological features of 50 cases.MethodsThe clinicopathological features of total 50 cases of FDCS were analyzed including a review of 44 cases reported in Chinese literature before October 2009 and six original cases from the pathology files conducted by the authors.ResultsThe youngest patient came under observation in this study is only seven years old. Including the cases contributed by the authors, our literary review indicated that male dominated the tumor cases (M: F = 3: 2). 28 cases (56%) present with this disease in extranodal sites. Tumor cells demonstrated positive staining for the follicular dendritic cell markers CD21 (47/49), CD35 (43/45), CD23 (20/23) and CD68 (23/25). In situ hybridization for Epstein-Barr virus-encoded RNA was performed in 10 cases. Nevertheless, EBV expression was absent in all these cases. The follow-up analysis of all cases shows that 26 (81.2%) patients were alive and disease free; 6 (18.8%) patients were alive with recurrent disease or metastasis; and nobody had died of this disease at the time of last follow-up.ConclusionsThe diagnosis of the FDCS is based on the findings of morphology and immunohistochemistry. The FDCS occurred in China should be viewed and treated as a low-grade sarcoma, and the role of the EBV in the pathogenesis of this tumor is still uncertain. There is a possibility that the tumor might be racial or geographic correlated, because most cases were reported from Eastern Asia area; its particular the case of the liver or spleen tumor.