miR-630 overexpression enhanced ionic radiation (IR)-induced cytotoxicity.

<p>(A) miR-630 significantly increased IR-induced inhibition rate (B and C) miR-630 strongly increased caspase 3 and caspase 6 activities following IR exposure (D and E) miR-630 significantly increased apoptosis rate after IR exposure. Error bars represent the mean of three separate determinations ± standard deviation (SD). Asterisk indicates statistically significant changes: * (P < 0.05), ** (P < 0.01).</p

TP53RK and BCL2L2 are direct targets of miR-630.

<p>(A) Predicted binding sites of miR-630 in the 3′-UTRs of TP53RK and BCL2L2 (B and C) Fluorescence was significantly reduced in the miR-630 mimics and wild-type dual-luciferase reporter plasmid-transfected group, while it showed no significant change in the miR-630 mimics and mutant dual-luciferase reporter plasmid-transfected group (D) TP53RK and BCL2L2 protein expression levels decreased in miR-630-transfected cells. Error bars represent the mean of three separate determinations ± standard deviation (SD). Asterisk indicates statistically significant changes: * (P < 0.05), ** (P < 0.01).</p

Inhibition of miR-630 led to decreased IR sensitivity.

<p>(A) Inhibition of miR-630 significantly decreased IR-induced inhibition rate (B and C) Inhibition of miR-630 strongly decreased caspase 3 and caspase 6 activities following IR exposure. Error bars represent the mean of three separate determinations ± standard deviation (SD). Asterisk indicates statistically significant changes: * (P < 0.05), ** (P < 0.01).</p

DNA methylation status and transcription factor CREB mediated miR-630 biogenesis.

<p>(A) 5-aza-CdR significantly upregulated miR-630 expression (B) CREB was predicted to bind to the 5′-UTR of miR-630 host gene ARIH1 (C) CREB within the ARIH1 promoter region induced luciferase activity and mutation of predicted CREB binding sites reversed its effect (D) ChIP confirmed the binding of CREB to the ARIH1 promoter. Error bars represent the mean of three separate determinations ± standard deviation (SD). Asterisk indicates statistically significant changes: * (P < 0.05), ** (P < 0.01).</p

miR-630 expression levels reduced after radiation.

<p>(A and B) Endogenous miR-630 expression and IR-induced inhibition rate of serial CRC cell lines (C) miR-630 level decrease after radiation compared with their initial level. Error bars represent the mean of three separate determinations ± standard deviation (SD). Asterisk indicates statistically significant changes: * (P < 0.05), ** (P < 0.01).</p

TP53RK and BCL2L2 suppress the effects of miR-630 overexpression.

<p>(A) TP53RK or BCL2L2 protein caused a significantly decreased IR-induced inhibition rate in miR-630 transfected cell lines. (B and C) TP53RK or BCL2L2 protein decreased the fold of caspase 3 and caspase 6 activities following IR exposure in miR-630 transfected cell lines. Error bars represent the mean of three separate determinations ± standard deviation (SD). Asterisk indicates statistically significant changes: * (P < 0.05), ** (P < 0.01).</p

Additional file 1: Figure S1. of Investigating the Heavy Metal Adsorption of Mesoporous Silica Materials Prepared by Microwave Synthesis

schematic diagram of the dissolution of silica fume. Table S1. Purification rate of silica fume. Figure S2. the N2 adsorption–desorption isotherm of the sample prepared with HCl at room temperature within 24 h. Table S2. Textural properties of the sample prepared with HCl at room temperature within 24 h. Figure S3. TEM image of c-MCM-41(40) (along the channel). Figure S4. TEM image of h-MCM-41(40). Table S3. Fitting parameters of Langmuir and Freundlich isotherms for the adsorption of Cu2+, Pb2+, and Cd2+ on c-MCM-41(40). Figure S5. The kinetics of Cu2+, Pb2+, and Cd2+ adsorption on c-MCM-41(40). Figure S6. Plot of kinetic model for the adsorption of Cu2+, Pb2+, and Cd2+, (A) pseudo-first-order kinetic model (B) pseudo-second-order kinetic model. (DOCX 1066 kb

FBX8 Acts as an Invasion and Metastasis Suppressor and Correlates with Poor Survival in Hepatocellular Carcinoma

<div><p>Background</p><p>F-box only protein 8 (FBX8), a novel component of F-box proteins, is lost in several cancers and has been associated with invasiveness of cancer cells. However, its expression pattern and role in the progression of hepatocellular carcinoma remain unclear. This study investigated the prognostic significance of FBX8 in hepatocellular carcinoma samples and analyzed FBX8 function in hepatocellular carcinoma cells by gene manipulation.</p><p>Methodology</p><p>The expression of FBX8 was detected in 120 cases of clinical paraffin-embedded hepatocellular carcinoma tissues, 20 matched pairs of fresh tissues and five hepatocellular carcinoma cell lines by immunohistochemistry with clinicopathological analyses, real-time RT-PCR or Western blot. The correlation of FBX8 expression with cell proliferation and invasion in five HCC cell lines was analyzed. Moreover, loss of function and gain of function assays were performed to evaluate the effect of FBX8 on cell proliferation, motility, invasion <i>in vitro</i> and metastasis <i>in vivo</i>.</p><p>Conclusions</p><p>We found that FBX8 was obviously down-regulated in HCC tissues and cell lines (P<0.05). The FBX8 down-regulation correlated significantly with poor prognosis, and FBX8 status was identified as an independent significant prognostic factor. Over-expression of FBX8 decreased proliferation, migration and invasion in HepG2 and 97H cells, while knock-down of FBX8 in 7721 cells showed the opposite effect. FBX8 negatively correlated with cell proliferation and invasion in 7701, M3, HepG2 and 97H cell lines. In vivo functional assays showed FBX8 suppressed tumor growth and pulmonary metastatic potential in mice. Our results indicate that down-regulation of FBX8 significantly correlates with invasion, metastasis and poor survival in hepatocellular carcinoma patients. It may be a useful biomarker for therapeutic strategy and control in hepatocellular carcinoma treatment.</p></div

FBX8 inhibits proliferation of HCC cells in vitro.

<p>(A) FBX8 expression in FBX8 overexpressing 97H and HepG2 cells by real-time RT-PCR. (B) FBX8 expression in FBX8 overexpressing 97H and HepG2 cells by Western blotting. (C) FBX8 expression in FBX8 depleting 7721 cells by real-time RT-PCR. (D) FBX8 expression in FBX8 depleting 7721 cells by Western blotting. (E) Effect of ectopic FBX8 on cell proliferation in vitro by MTT assay. (F) Effect of FBX8 knockdown on cell proliferation in vitro by MTT assay. (G) Effect of ectopic FBX8 on cell proliferation in vitro by colony formation assay. *P<0.05.</p

FBX8 is down-regulated in HCC cell lines and fresh HCC tissues.

<p>(A) Kaplan-Meier survival analysis of primary HCC patients with high and low FBX8 expressions. (B) Real-time RT-PCR analysis of FBX8 in six cell lines. The relative mRNA levels with the use of control LO2 were normalized to 1. (C) Western blotting analysis of FBX8 in six cell lines. (D) Real-time PCR analysis of FBX8 expression in 8 paired HCC tissues. (E) Western blotting analyses of FBX8 expression in the same 8 paired HCC tissues. N = normal mucosa and T = tumor.</p