Long non-coding RNA PVT1 facilitates cell migration and invasion by regulating miR-148a-3p and ROCK1 in breast cancer.

PURPOSE: Breast cancer (BC) is one of the most common malignant tumors for women. The role and potential mechanisms of long non-coding RNA plasmacytoma variant translocation 1 (lncRNA PVT1) were explored in BC cell migration and invasion. METHODS: PVT1, miR-148a-3p and Rho­associated, coiled­coil containing protein kinase 1 (ROCK1) mRNA expressions were detected using real-time fluorescent quantitative polymerase chain reaction (qRT-PCR). The ROCK1 protein expression was detected by Western blotting. The relationship of PVT1, miR-148a-3p and ROCK1 was analyzed by Dual Luciferase activity, RNA immunoprecipitation (RIP) and Spearman correlation analysis. Cell invasion and migration were detected by Transwell assay. RESULTS: Upregulation of PVT1 and ROCK1, and downregulation of miR-148a-3p were observed in BC tissues and cell lines. According to the analysis of Dual Luciferase activity, RIP and Spearman correlation analysis, miR-148a-3p directly binds to PVT1, and ROCK1 is a target of miR-148a-3p. In addition, PVT1 regulated the cells migration and invasion by regulating miR-148a-3p and ROCK1 expression. CONCLUSION: These data demonstrated that PVT1 was upregulated and facilitated to the cell migration and invasion of BC by the regulation of miR-148a-3p and ROCK1, indicating that PVT1 may be a potential biomarker of BC diagnosis and treatment.

Influence of suppression of CapG gene expression by siRNA on the growth and metastasis of human prostate cancer cells.

This study investigated CapG gene expression in prostate cancer cell lines; in addition, we explored the effects of CapG suppression on DU145 cell growth, and the underlying mechanism with which CapG affects DU145 cell growth and invasiveness. The expression of CapG and 18 related genes in DU145 cells was analyzed by flow cytometry, quantitative polymerase chain reaction (qPCR), CCK8 assay, western blot, and the trans-well assay. DU145 cells were transfected with designed small interfering RNA (siRNA). CapG expression was quantified by qPCR and western blot. DU145 cell proliferation and invasiveness was analyzed using the CCK8, flow cytometric, and trans-well assays. CapG, TMPRSS1, EGFR, ETS-1, ERBB2, AKT, Cyclin D1, P21, Bcl-2, and Bak1 gene and Bcl-2, Cyclin D1, and CapG protein expressions were significantly lower in the siRNA group compared to the negative control group (P < 0.05). The proliferation of CapG siRNA DU145 cells was lower than that of the two control groups, 48 h after transfection. The cell inhibition rate was 24.5, 35.4, and 16,5% at 24, 48, and 72 h, respectively. The growth curve indicated that CapG siRNA DU145 cells showed a significantly slower proliferation rate (P < 0.05). The trans-well assay showed a significant decrease in the migratory and invasive capacities of DU145 cells in the siRNA group (P < 0.05). The suppression of CapG expression caused a significant decrease in the proliferation, invasiveness, and metastasis of DU145 cells. The mechanism with which CapG, with other oncogenes, influences cancer cell cycle remains to be elucidated.

Significance of sarcomere gene mutation in patients with dilated cardiomyopathy.

Dilated cardiomyopathy (DCM) is a myocardial disease with a high mortality rate. Approximately 40 genes have been found to be associated with DCM to date. Non-familial DCM can also be caused by gene mutations, suggesting that genetic factors were involved in the pathogenesis of DCM; therefore genetic testing is beneficial for the early diagnosis of DCM, which can facilitate the implementation of preventive measures by and within patient's families. Here, we investigated the underlying genetic mutations involved in the cause of patients with DCM. This prospective study included 240 patients with idiopathic DCM and 240 healthy volunteers. Subject clinical data were collected and polymerase chain reaction amplification was carried out on subject DNA for three candidate genes tropomyosin (TPM1), cardiac troponin T type-2 (TNNT2), and nuclear lamina protein A/C. Single nucleotide polymorphism (SNP) loci were detected in the TPM1 (rs1071646) and TNNT2 (rs3729547) genes, respectively. The genotype distributions and allele frequencies were found to satisfy Hardy-Weinberg equilibrium, which indicated that the group was representative. Statistically significant differences were found between the variant frequencies in the two SNP loci between the Kazakh patients with idiopathic DCM (IDCM) and healthy volunteers. A significant difference in the genotype distributions (P = 0.000) and allele frequencies (P = 0.000) of SNP rs1071646, and another significant difference in the genotype distributions (P = 0.000) and allele frequencies (P = 0.039) of SNP rs3729547 between Kazakhs with IDCM and Kazakh controls. These results suggest that the TPM1 (rs1071646) and TNNT2 (rs3729547) gene variants might represent risk factors for patients with DCM in the Kazakh population.

Effects of dietary crude protein on the growth performance, carcass characteristics and serum biochemical indexes of Lueyang black-boned chickens from seven to twelve weeks of age

This study was undertaken to assess dietary crude protein (CP) concentration for optimum growth performance and carcass characteristics of Lueyang black-boned chicken. In total, six hundred 42-day-old Lueyang black-boned chicks were randomly assigned to five treatments, each with six replicate pens with ten males and ten females. The birds fed experimental diets with different levels of protein concentration of 120, 140, 160, 180 and 200 g kg-1 from seven to twelve weeks of age respectively. On day of 84, weight gain, feed intake, and feed:gain ratio were measured, and two chickens (one male and one female) close to the average weight of all birds in each treatment were selected from each pen and sacrificed to evaluate carcass traits and selected serum biochemical indexes. Dietary CP concentration did not have any significant influence on feed intake (p>0.05). The birds fed the diet with 180 or 160 g kg-1 CP concentration exhibited greater (p< 0.05) growth rate, better feed conversion ratio, relative breast weight and albumin concentration in serum than that of those fed other dietary CP concentrations. According to the results of regression analysis, the CP requirements of Lueyang black-boned chicken from seven to twelve weeks of age for optimal weight gain and feed:gain ratio were 174 and 170 g kg-1, respectively.

Effects of dietary crude protein on the growth performance, carcass characteristics and serum biochemical indexes of Lueyang black-boned chickens from seven to twelve weeks of age

This study was undertaken to assess dietary crude protein (CP) concentration for optimum growth performance and carcass characteristics of Lueyang black-boned chicken. In total, six hundred 42-day-old Lueyang black-boned chicks were randomly assigned to five treatments, each with six replicate pens with ten males and ten females. The birds fed experimental diets with different levels of protein concentration of 120, 140, 160, 180 and 200 g kg-1 from seven to twelve weeks of age respectively. On day of 84, weight gain, feed intake, and feed:gain ratio were measured, and two chickens (one male and one female) close to the average weight of all birds in each treatment were selected from each pen and sacrificed to evaluate carcass traits and selected serum biochemical indexes. Dietary CP concentration did not have any significant influence on feed intake (p>0.05). The birds fed the diet with 180 or 160 g kg-1 CP concentration exhibited greater (p< 0.05) growth rate, better feed conversion ratio, relative breast weight and albumin concentration in serum than that of those fed other dietary CP concentrations. According to the results of regression analysis, the CP requirements of Lueyang black-boned chicken from seven to twelve weeks of age for optimal weight gain and feed:gain ratio were 174 and 170 g kg-1, respectively.(AU)

Cassava (Manihot esculenta Krantz) genome harbors KNOX genes differentially expressed during storage root development.

In plants, homeodomain proteins play a critical role in regulating various aspects of plant growth and development. KNOX proteins are members of the homeodomain protein family. The KNOX transcription factors have been reported from Arabidopsis, rice, and other higher plants. The recent publication of the draft genome sequence of cassava (Manihot esculenta Krantz) has allowed a genome-wide search for M. esculenta KNOX (MeKNOX) transcription factors and the comparison of these positively identified proteins with their homologs in model plants. In the present study, we identified 12 MeKNOX genes in the cassava genome and grouped them into two distinct subfamilies based on their domain composition and phylogenetic analysis. Furthermore, semi-quantitative reverse transcription polymerase chain reaction analysis was performed to elucidate the expression profiles of these genes in different tissues and during various stages of root development. The analysis of MeKNOX expression profiles of indicated that 12 MeKNOX genes display differential expressions either in their transcript abundance or expression patterns.

SAHA attenuates sevoflurane-induced learning and memory impairments in fetal mice.

Previous studies have found that children with multiple exposures to anesthesia at an early age are at increased risk of learning and memory impairment. Sevoflurane is the most commonly used inhalational anesthetic for general anesthesia in children. Multiple exposures to sevoflurane have been shown to induce neuroinflammation, inhibit neurogenesis, and cause subsequent learning and memory impairments in fetal mice. Histone-tail acetylation has been implicated in memory formation. In this study, we employed suberanilohydroxamic acid (SAHA), an inhibitor of histone deacetylases, to treat sevoflurane-induced learning and memory impairments. Six-day-old C57BL/6 mice were exposed to sevoflurane for 2 h daily for 3 days. Morris water maze test performed to evaluate learning and memory impairments and the expression of genes related in to synaptic remodeling/plasticity, or regulated by neuronal activity or the cell cycle were detected by real-time PCR. We found that SAHA attenuated sevoflurane-induced learning and memory impairments in fetal mice. Our findings suggest that SAHA may have potential as a therapeutic agent for preventing or treating the neurotoxicity associated with anesthesia.

Cloning and molecular characterization of a cDNA encoding a small GTPase from Hevea brasiliensis.

Small GTPases play a critical role in the regulation of a range of cellular processes including growth, differentiation, and intracellular transportation. The cDNA encoding a small GTPase, designated as HbGTPase1, was isolated from Hevea brasiliensis. HbGTPase1 was 882 bp long containing a 612-bp open reading frame encoding a putative protein of 203 amino acids, flanked by an 83-bp 5'-untranslated region (UTR) and a 187-bp 3'-UTR. The predicted molecular mass of HbGTPase1 is 22.62 kDa, with an isoelectric point of 5.06. The HbGTPase1 protein was predicted to possess the conserved functional regions of the small GTPase superfamily of proteins. Quantitative polymerase chain reaction analysis revealed that HbGTPase1 was constitutively expressed in all tissues tested. HbGTPase1 transcripts accumulated at relatively low levels in the flower, latex, and leaves, while HbGTPase1 transcripts accumulated at relatively high levels in bark. Transcription of HbGTPase1 in the latex was induced by jasmonate.

Resistance to ethanol disordering of membranes from ethanol-fed rats is conferred by all phospholipid classes.

Phospholipids extracted from liver microsomes and mitochondria of ethanol-fed rats retained the resistance to membrane disordered by ethanol which is observed in the intact isolated membranes. The lipid extracts were separated into the major phospholipid classes (phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol from microsomes and phosphatidylcholine, phosphatidylethanolamine and cardiolipin from mitochondria) by preparative TLC. The extent of membrane disordering by ethanol of phospholipid vesicles composed of a mixture of phospholipids from ethanol-fed rats and controls was determined from the reduction of the order parameter of the spin-probe 12-doxyl-stearate. In contrast to previous reports, we found that all phospholipid classes from ethanol-fed rats confer resistance to disordering by ethanol. To a first approximation the extent of resistance was proportional to the fraction of lipids from ethanol-fed rats, regardless of the phospholipid head-group. Subtle differences between phospholipid classes may exist but were too small to measure accurately. Except for phosphatidylethanol, incorporation of anionic phospholipids did not have a significant effect on the sensitivity of phospholipid vesicles to the disordering effect of ethanol. Vesicles prepared from mixtures of various dioleoyl phospholipids and natural phospholipids did not indicate a clear effect of fatty acid saturation on the sensitivity to disordering by ethanol. Although the precise molecular changes that occur in phospholipids from ethanol-fed rats have not been fully characterized it appears that subtle changes in all phospholipid classes contribute to the resistance to ethanol disordering of these membranes.