Background: Metastasis is a major reason for poor prognosis in patients with cancer, including hepatocellular carcinoma (HCC). A salient feature is the ability of cancer cells to colonize different organs. Long non-coding RNAs (lncRNAs) play important roles in numerous cellular processes, including metastasis. Materials and Methods: In this study, the lncRNA expression profiles of two HCC cell lines, one with high potential for metastasis to the lung (HCCLM3) and the other to lymph nodes (HCCLYM-H2) were assessed using the Arraystar Human LncRNA Array v2.0, which contains 33,045 lncRNAs and 30,215 mRNAs. Coding-non-coding gene co-expression (CNC) networks were constructed and gene set enrichment analysis (GSEA) was performed to identify lncRNAs with potential functions in organ-specific metastasis. Levels of two representative lncRNAs and one representative mRNA, RP5-1014O16.1, lincRNA-TSPAN8 and TSPAN8, were further detected in HCC cell lines with differing metastasis potential by qRT-PCR. Results: Using microarray data, we identified 1,482 lncRNAs and 1,629 mRNAs that were differentially expressed (${\geq}1.5$ fold-change) between the two HCC cell lines. The most upregulated lncRNAs in H2 were RP11-672F9.1, RP5-1014O16.1, and RP11-501G6.1, while the most downregulated ones were lincRNA-TSPAN8, lincRNA-CALCA, C14orf132, NCRNA00173, and CR613944. The most upregulated mRNAs in H2 were C15orf48, PSG2, and PSG8, while the most downregulated ones were CALCB, CD81, CD24, TSPAN8, and SOST. Among them, lincRNA-TSPAN8 and TSPAN8 were found highly expressed in high lung metastatic potential HCC cells, while lowly expressed in no or low lung metastatic potential HCC cells. RP5-1014O16.1 was highly expressed in high lymphatic metastatic potential HCC cell lines, while lowly expressed in no lymphatic metastatic potential HCC cell lines. Conclusions: We provide the first detailed description of lncRNA expression profiles related to organ-specific metastasis in HCC. We demonstrated that a large number of lncRNAs may play important roles in driving HCC cells to metastasize to different sites; these lncRNAs may provide novel molecular biomarkers and offer a new basis for combating metastasis in HCC cases.
Alhazmi, Mohammed I.;Hasan, Tarique N.;Shafi, Gowhar;Al-Assaf, Abdullah H.;Alfawaz, Mohammed A.;Alshatwi, Ali A.
Asian Pacific Journal of Cancer Prevention
/
v.15
no.22
/
pp.9655-9660
/
2014
Background: Nigella Sativa (NS) is an herb from the Ranunculaceae family that exhibits numerous medicinal properties and has been used as important constituent of many complementary and alternative medicines (CAMs). The ability of NS to kill cancer cells such as PC3, HeLa and hepatoma cells is well established. However, our understanding of the mode of death caused by NS remains nebulous. The objective of this study was to gain further insight into the mode and mechanism of death caused by NS in breast cancer MCF-7 cells. Materials and Methods: Human breast cancer cells (MCF-7) were treated with a methanolic extract of NS, and a dose- and time-dependent study was performed. The $IC_{50}$ was calculated using a Cell Titer $Blue^{(R)}$ viability assay assay, and evidence for DNA fragmentation was obtained by fluorescence microscopy TUNEL assay. Gene expression was also profiled for a number of apoptosis-related genes (Caspase-3, -8, -9 and p53 genes) through qPCR. Results: The $IC_{50}$ of MCF-7 cells was $62.8{\mu}L/mL$. When MCF-7 cells were exposed to $50{\mu}L/mL$ and $100{\mu}L/mL$ NS for 24h, 48h and 72h, microscopic examination (TUNEL assay) revealed a dose- and time-dependent increase in apoptosis. Similarly, the expression of the Caspase-3, -8, -9 and p53 genes increased significantly according to the dose and time. Conclusions: NS induced apoptosis in MCF-7 cells through both the p53 and caspase pathways. NS could potentially represent an alternative source of medicine for breast cancer therapy.
Background: The discovery that microRNAs (miRNAs) regulate proliferation, invasion and metastasis provides a principal molecular basis of tumor heterogeneity. Microvessel distribution is an important characteristic of solid tumors, with significant hypoxia occurring in the center of tumors with low blood flow. The distribution of miR-374a in breast tumors was examined as a factor likely to be important in breast cancer progression. Methods: Breast tissue samples from 40 patients with breast cancer were classified into two groups: a highly invasive and metastatic group (HIMG) and a low-invasive and metastatic Group (LIMG). Samples were collected from the center and edge of each tumor. In each group, six specimens were examined by microRNA array, and the remaining 14 specimens were used for real-time RT-qPCR, Western blot and immunohistochemical analyses. Correlation analysis was performed for the miRNAs and target proteins. Follow-up was carried out during 28 months to 68 months after surgery, and survival data were analyzed. Results: In the LIMG, the relative content of miR-374a was lower in the center of the tumor than at its edge; in the HIMG, it was lower at the edge of the tumor, and miR-374a levels were lower in breast cancer tissues than in normal tissues. There was no difference between VEGF-A and VCAM-1 mRNA levels at the edge and center of the tumor; however, we observed a significant difference between VEGF-A and VCAM-1 protein expression levels in these two regions. There was a negative correlation between miR-374a and target protein levels. The microvessel density (MVD) was lower in the center of the tumor than at its edge in HIMG, but the LIMG vessels were uniformly distributed. There was a significant positive correlation between MVD and the number of lymph node metastases (Pearson correlation, r=0.912, P<0.01). The median follow-up time was 48.5 months. LIMG had higher rate of disease-free survival (100%, P=0.013) and longer median survival time (66 months) than HIMG, which had a lower rate of 75% and shorter median survival time (54 months). Conclusions: Our data demonstrated miR-374a to be differentially distributed in breast cancer; VEGF-A and VCAM-1 mRNA had coincident distribution, and the distribution of teh respective proteins was uneven and opposite to that for the miR-374a. These data might explain the differences in the distribution of MVD in breast cancer and variation in breast cancer prognosis.
The 7F0→5D0excitation spectra of Eu(Ⅲ) complexed with polyfunctional monocarboxylic acid(glycolic acid, glycine and thioglycolic acid) containing a terminal O, N and S neutral donors and propionic acid were investigated using Eu(Ⅲ) luminescence spectroscopy. In the excitation spectra of Eu(Ⅲ)-propionate system, the stepwise appearance of the peaks was observed at 579.0, 579.2 and 579.5 nm with increasing in the ligand-to-metal ratio, which correspond to the formation of Eu(propionate)2+, Eu(propionate)2+ and Eu(propionate)3 species. Three maximum peaks were also obtained for Eu(Ⅲ)-glycolate, Eu(Ⅲ)-glycinate and Eu(Ⅲ)-thioglycolate systems and were found to be quite similar to those of Eu(Ⅲ)-propionate system. The q values (number of coordinated water molecules of Eu(Ⅲ) ion) obtained from the luminescence decay constants of Eu(Ⅲ)-glycolate and Eu(Ⅲ)-thioglycolate were 7.0 and 7.1, and compare well with 7.3 for Eu(Ⅲ)-propionate: Each ligand units replace around two coordinated water molecules. These results show that the polyfunctional monocarboxylates behaves like the propionate for Eu(Ⅲ) ion coordination.
In order to provide basic data for the ecological management of forest vegetation in Southern Naeyeon Mountains, A total of 149 sample plots were selected and vegetation survey was carried out by the phytosociological method of the ZM school to classify vegetation types and to grasp ecological characteristics. The forest vegetation was divided into 10 types in terms of species composition, and had a unit hierarchy of 2 community groups, 4 communities, 6 sub-communities and 6 variants. A total of 19 types of physiognomic vegetation were identified based on uppermost dominant species, of which 18 were natural vegetation and 1 was artificial vegetation. As a result of the analysis of the importance values of constituent species, Quercus mongolica, a potentially natural vegetation element, was found to be relatively more important in most stands than other species, and excluding the artificial interference, most of the areas except for some sites would be changed to Q. mongolica forest. In order to understand the spatial distribution of forest vegetation, 1/5,000 large-scale physiognomic vegetation map was created by the uppermost dominant species. As a result, natural vegetation accounted for 98.2%, the number of vegetation patches was 733 and the average area per patch 3.93ha.
This study was conducted to investigate the effect of different pre-sowing treatments of seeds on germination and growth performance of native threatened tree species Quercus gomeziana A. Camus at the nursery of Chittagong University, Bangladesh. Furthermore, seedling growth attributes under different doses of fertilizer (urea) was also experimented to find the best dose of fertilizer on this tree species at the nursery stage for better field level growth. Seeds were placed to six pre-sowing treatments e.g. control (PT0), treated with sand paper rubbing (PT1), nicking (PT2), seeds immersed in cold water for 48 hours (PT3), seeds immersed in cold water for 7 days (PT4) and seeds sown at propagator house with increased temperature (PT5). It was found from the study that germination was started earlier (at 31 days) in treatments sand paper rubbing (PT1) and nicking (PT2). The highest germination percentage (93%) was in PT1 followed by 86% in seeds immersed in cold water for 7 days (PT4) and 80% in PT0 (control). Germination percentage was observed least (63%) in PT2 even though germination started earlier. For fertilizer dose experiment to seedlings at the nursery level, treatment FT1: 100 kg/ha (0.33679 g urea/pot/seedling) comparing with other treatments FT0: 0 kg/ha (Control), FT2: 200 kg/ha (0.67358 g urea/pot/seedling), FT3: 300 kg/ha (1.01037 g urea/pot/seedling) showed better performance in case of collar diameter (6.74 mm), number of leaves, shoot dry weight (19.74), total dry weight (28.16 g), total fresh weight (67.96 g), volume index (3904.82), sturdiness (127.69). Finally, it can be concluded that Quercus gomeziana seedlings revealed better performances under the treatment FT1 in growth and biomass production. Findings of this study will be helpful to take decision on organic fertilizer dose application to seedlings of Q. gomeziana for large scale plantation and conservation of this species.
Utilizing chi-square test statistics, inter-species association and covariation were analyzed for the 37 woody plant species in a deciduous forest dominated by Quercus mongolica and Q. variabilis. within 50 temporarily established $20m{\times}20m$ square quadrats, the association for each pair of species was presented based on the presence-absence parameters. Acer palmatum had significant positive association with Acer mono and Kalopanax pictus, but negative association with Pinus densiflora. Other positively associated species pairs were Prunus sargentii-Macckia amurensis, Quercus serrata-Kalopanax pictus, Symplocos chinensis var. pilosa-Euonymus oxyphyllus, and Ulmus davidiana var. japonica-Lindera obtusiloba. The covariation far each pair of species was evaluated based on the quantitative measures, density and basal area. Overall results showed that the association and covariation values among species generally agreed with each other. Because covariation was calculated by density and basal area of the tallied species in the sample plots, the number of species pairs of covariation tended to be greater than those of association. Especially, Pinus densiflora, considered to be pioneer species in the successional stage, had negative covariation with most of climax species. These ecological information could be applied to silvicultural practices, such as ecosystem classification, establishment of mixed hardwood forest, and tending operations for marking crop trees and desirable species.
In this work, small-size, high-performance solenoid-type RF chip inductors utilizing a low-loss ${Al_2}{O_3}$ core material were investigated. The size of the chip inductors fabricated in this work were $0.86{\times}0.46{\times}0.45m^3$, $1.5{\times}1.0{\times}0.7m^3$, $2.1{\times}1.5{\times}1.0m^3$, and $2.4{\times}2.0{\times}1.4m^3$ and copper (Cu) wire with $27{\sim}40{\mu}m$ diameter was used as the coils. High frequency characteristics of the inductance, quality factor, and impedance of developed inductors were measured using an RF Impedance/Material Analyzer (HP4291B with HP16193A test fixture). It was observed that the developed inductors with the number of turns of 7 have the inductance of 13 to 100nH and exhibit the self-resonant frequency (SRF) of 6.4 to 1.1GHz. The SRF of inductors decreases with increasing the inductance and the inductors have the quality factor of 50 to 80 in the frequency range of 300MHz to 1.3GHz. In this study, small-size solenoid-type RF chip inductors with high inductance and high quality factor were fabricated successfully.
This trial was conducted to determine the effects of feeding a diet containing solid-state fermented rapeseed meal on performance, nutrient digestibility, intestinal ecology and intestinal morphology of broiler chickens. A mixed liquid culture, containing approximately 5 log cfu/ml Lactobacillus fermentum, Enterococcus faecium, Saccharomyces cerevisae and Bacillus subtilis was prepared in a 1:1:1:1 ratio. A basal substrate (BS) containing 75% rapeseed, 24% wheat bran and 1% brown sugar was mixed with the liquid culture in a ratio of 10:3. Over the 30-day fermentation, isothiocyanates were reduced from 119.6 to 14.7 mmol/kg. A total of 168, day-old male Arbor Acres broiler chicks were assigned to one of three dietary treatments including a corn-soybean meal based control diet as well as two experimental diets in which the control diet was supplemented with 10% of the BS containing unfermented rapeseed meal or 10% of the BS containing rapeseed meal subjected to solid state fermentation. There were 8 pens per treatment and 7 birds per pen. From days 19-21 and days 40-42, uncontaminated excreta were collected from each pen for digestibility determinations. In addition, digesta from the colon and ceca were collected to determine the number of lactobacilli, enterobacteria and total aerobes. The middle sections of the duodenum, jejunum, and ileum were collected for intestinal morphology. Over the entire experimental period (d 1-42), the weight gain and feed conversion of birds fed fermented rapeseed meal were superior (p<0.05) to that of birds fed nonfermented rapeseed meal and did not differ from the soybean control. On day 42, birds fed fermented rapeseed meal had higher (p<0.05) total tract apparent digestibility coefficients for dry matter, energy, and calcium than birds fed non-fermented rapeseed meal. Colon and ceca digesta from broilers fed the fermented feed had higher (p<0.05) lactobacilli counts than birds fed the control and non-fermented rapeseed meal diets on day 21 and 42. Fermentation also improved (p<0.05) villus height and the villus height:crypt depth ratio in the ileum and jejunum on day 21 and 42. The results indicate that solid-state fermentation of rapeseed meal enhanced performance and improved the intestinal morphology of broilers and may allow greater quantities of rapeseed meal to be fed to broilers potentially reducing the cost of broiler production.
Background: Aberrant promoter hypermethylation has been recognized in human breast carcinogenesis as a frequent molecular alteration associated with the loss of expression of a number of key regulatory genes and may serve as a biomarker. The E-cadherin gene (CDH1), mapping at chromosome 16q22, is an intercellular adhesion molecule in epithelial cells, which plays an important role in establishing and maintaining intercellular connections. The aim of our study was to assess the methylation pattern of CDH1 and to correlate it with the expression of E-cadherin, clinicopathological parameters and hormone receptor status in breast cancer patients of Kashmir. Materials and Methods: Methylation specific PCR (MSP) was used to determine the methylation status of CDH1 in 128 invasive ductal carcinomas (IDCs) paired with the corresponding normal tissue samples. Immunohistochemistry was used to study the expression of E-cadherin, ER and PR. Results: CDH1 hypermethylation was detected in 57.8% of cases and 14.8% of normal adjacent controls. Reduced levels of E-cadherin protein were observed in 71.9% of our samples. Loss of E-cadherin expression was significantly associated with the CDH1 promoter region methylation (p<0.05, OR=3.48, CI: 1.55-7.79). Hypermethylation of CDH1 was significantly associated with age at diagnosis (p=0.030), tumor size (p=0.008), tumor grade (p=0.024) and rate of node positivity or metastasis (p=0.043). Conclusions: Our preliminary findings suggest that abnormal CDH1 methylation occurs in high frequencies in infiltrating breast cancers associated with a decrease in E-cadherin expression. We found significant differences in tumor-related CDH1 gene methylation patterns relevant to tumor grade, tumor size, nodal involvement and age at diagnosis of breast tumors, which could be extended in future to provide diagnostic and prognostic information.
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