Minicircle-oriP-IFNγ: A Novel Targeted Gene Therapeutic System for EBV Positive Human Nasopharyngeal Carcinoma

) in which the transgene expression was under the transcriptional regulation of oriP promoter.. Immunohistochemistry was used to detect the expression and the activity of the IFNγ in tumor sections. Our results demonstrated that mc-oriP vectors mediated comparable gene expression and anti-proliferative effect in the EBV-positive NPC cell line C666-1 compared to mc-CMV vectors. Furthermore, mc-oriP vectors exhibited much lower killing effects on EBV-negative cell lines compared to mc-CMV vectors. The targeted expression of mc-oriP vectors was inhibited by EBNA1-siRNA in C666-1. This selective expression was corroborated in EBV-positive and -negative tumor models. as a safe and highly effective targeted gene therapeutic system for the treatment of EBV positive NPC

Cryptochrome 1 overexpression correlates with tumor progression and poor prognosis in patients with colorectal cancer.

BACKGROUND: Clock genes drive about 5-15% of genome-wide mRNA expression, and disruption of the circadian clock may deregulate the cell's normal biological functions. Cryptochrome 1 is a key regulator of the circadian feedback loop and plays an important role in organisms. The present study was conducted to investigate the expression of Cry1 and its prognostic significance in colorectal cancer (CRC). In addition, the function of Cry1 in human CRC was investigated in cell culture models. METHODS: Real-time quantitative PCR, Western blot analysis and immunohistochemistry were used to explore Cry1 expression in CRC cell lines and primary CRC clinical specimens. MTT and colony formation assays were used to determine effects on cellular proliferation ability. The animal model was used to explore the Cry1 impact on the tumor cellular proliferation ability in vivo. Transwell assays were performed to detect the migration ability of the cell lines. Statistical analyzes were applied to evaluate the diagnostic value and the associations of Cry1 expression with clinical parameters. RESULTS: Cry1 expression was up regulated in the majority of the CRC cell lines and 168 primary CRC clinical specimens at the protein level. Clinical pathological analysis showed that Cry1 expression was significantly correlated with lymph node metastasis (p = 0.004) and the TNM stage (p = 0.003). High Cry1 expression was associated with poor overall survival in CRC patients (p = 0.010). Experimentally, we found that up-regulation of Cry1 promoted the proliferation and migration of HCT116 cells, while down-regulation of Cry1 inhibited the colony formation and migration of SW480 cells. CONCLUSIONS: These results suggest that Cry1 likely plays important roles in CRC development and progression andCry1 may be a prognostic biomarker and a promising therapeutic target for CRC

Long Noncoding RNA BC032913 as a Novel Therapeutic Target for Colorectal Cancer that Suppresses Metastasis by Upregulating TIMP3

Summary: Long noncoding RNAs (lncRNAs) have been shown to play critical roles in the biology of various cancers. However, their expression patterns and biological functions in human colorectal cancer (CRC) remain largely unknown. The aim of this study was to explore lncRNA profiles in CRC and investigate key lncRNAs involved in CRC tumorigenesis and progression. The microarray data of six CRC and matched non-cancerous tissues revealed distinct lncRNA profiles, including 899 upregulated and 1,646 downregulated lncRNAs (p  2.0). Furthermore, we found that the lncRNA BC032913 was generally underexpressed in 115 CRC samples compared with normal tissues. Reduced BC032913 levels were significantly associated with an advanced tumor, lymph nodes, distant metastasis (TNM) stage and a higher risk of lymph node and distant metastases. BC032913 downregulation indicated poor overall survival in CRC patients. Moreover, BC032913 enhanced the mRNA and protein expression of TIMP3 and inhibited Wnt/β-catenin pathway activity, thus suppressing CRC metastasis in vitro and in vivo. Collectively, the obtained data show that BC032913 plays an inhibitory role in CRC aggression by upregulating TIMP3, followed by inactivation of the Wnt/β-catenin pathway. Our findings indicate that the novel lncRNA BC032913 could serve as a novel prognostic marker and effective therapeutic target for CRC. Keywords: colorectal cancer metastasis, lncRNA BC032913, TIMP

Extracellular-vesicle-packaged S100A11 from osteosarcoma cells mediates lung premetastatic niche formation by recruiting gMDSCs

Summary: The premetastatic niche (PMN) contributes to lung-specific metastatic tropism in osteosarcoma. However, the crosstalk between primary tumor cells and lung stromal cells is not clearly defined. Here, we dissect the composition of immune cells in the lung PMN and identify granulocytic myeloid-derived suppressor cell (gMDSC) infiltration as positively associated with immunosuppressive PMN formation and tumor cell colonization. Osteosarcoma-cell-derived extracellular vesicles (EVs) activate lung interstitial macrophages to initiate the influx of gMDSCs via secretion of the chemokine CXCL2. Proteomic profiling of EVs reveals that EV-packaged S100A11 stimulates the Janus kinase 2/signal transducer and activator of transcription 3 signaling pathway in macrophages by interacting with USP9X. High level of S100A11 expression or circulating gMDSCs correlates with the presentation of lung metastasis and poor prognosis in osteosarcoma patients. In summary, we identify a key role of tumor-derived EVs in lung PMN formation, providing potential strategies for monitoring or preventing lung metastasis in osteosarcoma

The overexpression of Cry1 mRNA and protein in colorectal cancer tissues.

<p>(<b>A</b>) A representative image of Cry1 staining in colorectal cancer tissues is shown. (<b>B</b>) A representative image of Cry1 staining in adjacent noncancerous tissues is shown. (<b>C</b>) Cry1 protein expression level was higher in tumor tissues compared to adjacent control tissue as detected by immunoblotting (mean±SEM; n = 109; * **, <i>P</i><0.001). (<b>D</b>) Average T/N ratios of Cry1 mRNA expression in paired colorectal cancer (T) and normalmucosa tissues (N) were quantified by qPCR and normalized to GAPDH. Error bars represent the standard deviation of the mean (SD) calculated from three parallel experiments. Magnification is ×200.</p

The expression of Cry1 protein in colorectal cancer sections.

<p>Representative immunohistochemical images of colorectal cancer tissue specimens indicating negative or weakly detectable Cry1 staining (<b>A</b> and <b>B</b>); moderate Cry1 staining (<b>C</b>); and strong Cry1 staining (<b>D</b>) are shown. Magnification is ×200 (<b>A, B, C</b> and <b>D</b>).</p

Overexpression of Cry1 mRNA and protein in colorectal cancer cell lines.

<p>Expression of Cry1 mRNA and protein in colorectal cancer cell lines (SW480, SW620, HT29, THC8307, and HCT116) and FHC were examined by qPCR (<b>A</b>) and Western blotting (<b>B</b>). Expression levels were normalized to GAPDH. Error bars represent the standard deviation of the mean (SD) calculated from three parallel experiments, *, <i>P</i><0.05.</p

Cry1 promoted CRC growth <i>in vivo</i>.

<p>(<b>A</b>) The expression of Cry1 was markedly increased in the stable cell line Cry1-HCT116 compared with control stable cell line ctrl-HCT116. GAPDH was used as an internal control. (<b>B</b>) Representative images of tumors derived from Cry1-HCT116 or ctrl-HCT116,following subcutaneous xenograft transplants in nude mice (<b>C</b>) Overexpression of Cry1 promoted colorectal cancer growth. Tumor cells were injected subcutaneously into nude mice. Mice were sacrificed after 4 weeks, and the volume of each tumor was measured every 4 days. Bars, ±SEM; *<i>P</i><0.05, ** <i>p</i> = 0.01.</p