MicroRNA-520a-3p suppresses non-small-cell lung carcinoma by inhibition of High Mobility Group Box 1 (HMGB1).

OBJECTIVE: Currently, pathogenesis of non-small cell lung carcinoma (NSCLC) is still unknown and the treatment is far from ideal. Therefore, we investigated the effect of inhibiting microRNA-520a-3p in NSCLC cells. MATERIALS AND METHODS: NCI-H157 cells were treated with microRNA-520a-3p analogs for 48 h, or microRNA-520a-3p analogs and its inhibitor, for a total of 48 h. Many tests were performed, including MTT, flow cytometry, wound healing assay and transwell assay. The tumor model was established, and HMGB1 mRNA was detected by RT-PCR. Protein levels of HMGB1, MMP-2, MMP-9, Bcl-2, Bax, and Caspase-3 were assessed by Western Blot. RESULTS: microRNA-520a-3p could significantly inhibit the proliferation, migration and invasion of NCI-H157 cells, inducing their apoptosis. microRNA-520a-3p inhibited tumor growth and decreased the mRNA and protein levels of HMGB1. Additionally, it decreased the Bcl-2/Bax ratio, MMP-2 and MMP-9 expression, and increased caspase-3 expression. CONCLUSIONS: microRNA-520a-3p exhibited an effective inhibition on NCI-H157 tumor growth likely by inhibiting HMGB1 expression.

TM4SF5-CTD-2354A18.1-miR-4697-3P may play a key role in the pathogenesis of gastric cancer.

AIMS: Our aim is to identify important lncRNAs and mRNAs which may play a key role in contributing to pathogenesis of gastric cancer. METHODS: Different LncRNAs and mRNAs are identified by microarray in gastric cancer tissue and corresponding normal tissues. The function and relationship of different LncRNAs and mRNAs is performed by GO analysis and Pathway analysis and made code-non-code network (CNC) by Pearson correlation coefficients (PCC). Then mRNA-miRNA relationship is predicted through mRNA-miRNA relationship software (http://www.targetscan.org). Lastly, mRNA-miRNA-LncRNA network is established for further research. RESULTS: The expression profiles of 3732 lncRNAs showed different expression (fold change (FC)≥2.0, p<0.05) in gastric cancer tissue and normal tissue and expression profiles of 3994 mRNAs also showed different expression (FC≥2.0, p<0.05) in gastric cancer and corresponding normal tissue. CONCLUSION: The expression of TM4SF5, CTD-2354A18.1 and miR-4697-3P is in balance at physiological conditions, however, the balance is disrupted by some situations, which may contribute to gastric cancer. GO analysis and Pathway analysis also showed TM4SF5 played an important role in proliferation, differentiation and apoptosis. Therefore, TM4SF5-miR-4697-3P- CTD-2354A18.1 may play a key role in the pathogenesis of gastric cancer (Tab. 2, Fig. 4, Ref. 30).

Research of inhibition of survivin in rat HSC-T6 cell by siRNA interference.

OBJECTIVE: To explore the effects and mechanisms of siRNA targeting survivin of inducing apoptosis in rat HSC-T6 cells. METHODS: The experiment was divided into blank group, pGPU6/GFP/Neo-shNC group and pGPU6/GFP/Neo-siRNA group. The siRNA was transfected into HSC-T6 cells mediated by LipofectamineTM2000 for 24 h, and then the efficiency of transfection was observed by fluorescence microscopy. After transfection for 48h, the expression of survivin mRNA and protein was assessed by reverse transcription-polymerase chain reaction (RT-PCR) method and Western-blot, and the form of cells was observed by microscopy. The apoptosis rate of HSC-T6 cells was measured by the flow cytometry with PI staining. The expression of caspase-3 protein was assessed by western-blot. RESULTS: The prominent apoptosis of the pGPU6/GFP/Neo-siRNA group by PI staining was high, there was significant difference comparing with blank group and pGPU6/GFP/Neo- shNC group (p<0.05). The expression of caspase-3 by Western-blot in pGPU6/GFP/Neo-siRNA group was high, there was significant difference comparing with blank group and pGPU6/GFP/Neo-shNC group (p>0.05). CONCLUSION: siRNA targeting survivin can inhibit the expression of survivin mRNA and protein in rat HSC-T6 cells. Expression of survivin is negative correlation with expression of caspase-3. siRNA targeting survivin may up-regulate expression of caspase-3 and increase apoptosis of rat HST-T6 (Fig. 6, Ref. 24).

Aberrant gene expression profiles in hepatocellular carcinoma detected by microdissection.

The aim of this study was to identify genomic aberrations in hepatocellular carcinoma (HCC) by using laser capture microdissection (LCM) combined with microarray analysis. Samples were procured by LCM from HCC and patient-matched normal liver tissue surgically resected from 4 patients. RNA was isolated from the samples and reverse transcribed into cDNA. After 2-cycle linear amplification and 2-color fluorescent labeling, the cRNA was hybridized onto a whole genome microarray. All genes expressed in the normal and HCC samples were counted and analyzed. Differentially expressed genes were identified and the top 10 up and downregulated genes (totally 20 genes) were further evaluated. LCM was able to accurately capture 50-200 cells from HCC and control tissues. The microarray spectrum showed satisfactory detection of HCC-enriched genes. A total of 1361 differentially expressed genes were identified, among which, 607 were upregulated and 754 were downregulated. Among the top 20 up and downregulated genes, 4 genes had not been documented in the literature as being differentially expressed in any tumors. Thus, LCM is an effective approach for identifying aberrantly expressed genes in HCC, and may lead to the discovery of biomarkers for diagnostic and therapeutic applications.

Acetate production by a coupled syntrophic acetogenesis with homoacetogenesis process: effect of sludge inoculum concentration.

The effect of sludge inoculum concentration on acetate (Ac) production by a novel coupled system was investigated. The coupled system consisted ofa syntrophic acetogenesis phase and a homoacetogenesis phase. The results showed that the inoculum concentration not only affected the acetate production at the syntrophic acetogenesis phase but also that produced at the homoacetogenesis phase. When the inoculum concentration in the syntrophic acetogenesis phase was 4 g l(-1), the acetate yield of the coupled system was found to be about 0.36 g Ac g(-1) glucose. In addition, when the inoculum concentration in the homoacetogenesis phase was 4 g l(-1), a maximum acetate production of 11.2 g l(-1) was obtained in the syntrophic acetogenesis phase and the yield of acetate was 0.39 g Ac g(-1) glucose. Analysis shows that the variation in acetate yield might be attributed to different metabolic pathways and the imbalance between hydrogen production and hydrogen consumption in the coupled system with different inoculum concentrations.

Study of gastric and gallbladder kinetics with real-time ultrasonography in cases of functional dyspepsia.

Gastric and gallbladder emptying in 113 patients with functional dyspepsia (FD) were evaluated by real-time ultrasonography (RUS) after a liquid-fat meal by the patients, and compared with 15 healthy volunteers. The results showed that in FD group 69 patients (61.06%) had delayed gastric emptying, and 28 patients (24.77%) had gallbladder hypokinesia. Among them both delayed gastric emptying and gallbladder hypokinesia were found in 11 patients (9.7%), 44 patients (38.93%) had normal gastric emptying and 85 patients (75.22%) had normal gallbladder emptying.

[Clinical study of erythromycin action on gallbladder motility in patients with non-ulcer dyspepsia].

Thirty-two patients of non-ulcer dyspepsia (NUD) with gallbladder hypokinesia treated by oral erythromycin administration 0.125g three times daily for two weeks. Before and after oral erythromycin administration, gallbladder volumes were determined by ultrasound, and plasma motilin concentration were determined by radioimmunoassay. The results showed that before and after oral erythromycin administration, maximal percentage emptying of gallbladder were 48.24 +/- 8.30ml vs 69.74 +/- 10.78ml (P < 0.01), plasma motilin were 361.28 +/- 87.92ng/L vs 394.97 +/- 134.27ng/L (P > 0.05). It is indicated that erythromycin could reduce fasting and postprandial residual gallbladder volumes and increases maximal percentage emptying of gallbladder. It suggests that erythromycin might have an agonist action on the motilin receptor as an agonist action on the motilin receptor as an exogenously administrated motilin.