Research Article

Table of Contents | Genet. Mol. Res. 2016 (2)

Displaying 1 - 15 of 482
Research Article

The aim of the research was to examine the expression level of microRNA221/222 (miR-221/222) in the serum of patients with type 2 diabetes mellitus (T2DM) who are also diagnosed with post-menopausal breast cancer. We aimed to evaluate the differences in microRNA expression in patients with T2DM alone, patients with post-menopausal breast cancer alone, and patients with both T2DM and post-menopausal breast cancer. We selected 20 cases from a healthy control group, 30 cases from the group of patients with T2DM and obesity, 30 cases from the group of the patients with post-menopausal breast cancer, and 30 cases from the group of patients with both T2DM and post-menopausal breast cancer. The expression of miR-221/222 in the serum of the patients with post-menopausal breast cancer was higher than that of T2DM patients (P

Genet. Mol. Res. 15(2): gmr7259
DOI: 10.4238/gmr.15027259
Research Article

Toll-like receptors (TLRs) can specifically identify pathogen-associated molecular patterns (PAMPs) by recognizing structural patterns in diverse microbial molecules, and can provide an effective defense against multiple microbial infectious. A variety of TLRs can be expressed on the surface of liver parenchymal as well as nonparenchymal cells. Kupffer cells are a type of hepatic nonparenchymal macrophage, and are positively associated with the severity of liver fibrosis. They play an important role in the synthesis and deposition of the extracellular matrix by upregulating the expression of tissue inhibitor of metalloproteinases and downregulating the activity of matrix metalloproteinases. Cirrhosis, a chronic diffuse lesion usually accompanying extensive liver fibrosis and nodular regeneration, is caused by liver parenchymal cells repeating injury-repair following reconstruction of organizational structure in the hepatic lobules. Hepatocellular carcinoma is caused by repeated and persistent chronic severe liver injury, and partial hepatocytes can eventually transform into hepatoma cells. Multiple TLRs such as TLR2, TLR3, TLR4, and TLR9, as well as other receptors, can be expressed in cirrhosis and hepatocellular carcinoma. About 53 and 85% of hepatocellular carcinoma patients frequently express TLR3 and TLR9, respectively. The chronic and repeated liver injury caused by alcohol, and HBV, HCV, or other pathogens can be recognized by TLRs through the PAMP pathway, which directly increases the risk for hepatic cirrhosis and hepatocellular carcinoma. In this review, we briefly present evidence that the novel cellular molecular mechanisms of TLRs may provide more information about new therapeutics targets of the anti-inflammatory immune response.

Genet. Mol. Res. 15(2): gmr7419
DOI: 10.4238/gmr.15027419
Research Article

We investigated the effect of microRNA-27b (miR-27b), a gene expression regulatory factor, on the expression of monocyte chemoattractant protein-1 (MCP1) stimulated by interleukin 17 (IL-17). After IL-17 had been added to H9C2 cardiomyocytes, an miR-27b mimic was transfected into the H9C2 cells. The mRNA expression levels of miR-27b and MCP1 in the H9C2 cells were detected by SYBR green I fluorescence quantitative reverse transcription polymerase chain reaction. Enzyme-linked immunosorbent assay was used to measure the expression levels of MCP1 protein in the H9C2 cells. The expression of MCP1 mRNA in the H9C2 cells began to increase 2 h after IL-17 stimulation, reached a peak at 4 h, and then decreased. The MCP1 protein level increased gradually in the 24 h following IL-17 stimulation. After transfection with the miR-27b mimic, the expression of miR-27b in the H9C2 cells significantly increased than that in the miRNA negative control group (P

Genet. Mol. Res. 15(2): gmr7784
DOI: 10.4238/gmr.15027784
Research Article

Prostate cancer is a common malignant tumor in males with an unclear pathogenic mechanism. As one epigenetic regulation mechanism, DNA methylation of the whole genome and specific gene(s) plays critical roles in pathogenesis, progression, diagnosis, and treatment of prostate cancer. The E-Cadherin gene is involved in cell metabolism and has been suggested to be related with malignancy of multiple tumors. This study investigated the correlation between E-Cadherin methylation and malignancy of prostate cancer. Gradient concentrations of 5-Aza-CdR (5, 10, and 20 mM) were used to treat the prostate cancer cell line (LNCaP), and mRNA level of E-Cadherin was detected by reverse transcription-polymerase chain reaction (RT-PCR). A total of 82 prostate cancer patients were recruited to detect the methylation status of the promoter region of the E-Cadherin gene by pyrophosphate sequencing. Real-time fluorescent quantitative PCR (qRT-PCR) was employed to determine mRNA levels of E-Cadherin. Methylation and mRNA levels of E-Cadherin were analyzed by the SPSS software. With elevated concentrations of 5-Aza-CdR, mRNA levels of E-Cadherin gradually increased. DNA methylation levels of tumor tissues were significantly elevated with increased Gleason score (P 0.05). DNA methylation level was negatively correlated with mRNA expression of the E-Cadherin gene. Methylation in tumor tissues was significantly higher than that in tumor adjacent tissues (P

Genet. Mol. Res. 15(2): gmr8046
DOI: 10.4238/gmr.15028046
Research Article

Pinelliae rhizoma is the dried tuber of Pinellia ternata (Thunb.) Breit., and has been used for thousands of years as a traditional Chinese medicine. However, its genomic background is little known. With the development of high-throughput genomic sequencing, it is now easy and cheap to obtain genomic information. In this study, 193,032,910 high-quality clean reads were generated using the Illumina Hiseq 2000 platform. A total of 53,544 unigenes were identified from the contigs assembled. Functional annotation analysis annotated 37,318, 27,697, 23,043, 22,869, 23,328, and 27,415 unigenes. KEGG analysis revealed that five pathways (169 genes) were associated with alkaloid synthesis, 201 unigenes were related to fatty acid biosynthesis (ko00061), and 133 unigenes were involved in the biosynthesis of unsaturated fatty acids (ko01040). In addition, 6703 simple sequence repeats were designed based on the unigene sequences for screening germplasm resources in the future. These data are a valuable resource for genomic studies on Pinellia plants.

Genet. Mol. Res. 15(2): gmr8062
DOI: 10.4238/gmr.15028062
Research Article

Melanocortin-4 receptor (MC4R) is associated with feed intake, growth, fatness, and carcass composition in many domestic animals. The aim of this study was to evaluate the association of single nucleotide polymorphisms (SNPs) in MC4R with milk production traits in water buffalo. Eight SNPs were identified by direct polymerase chain reaction sequencing of samples from 18 buffaloes. SNPs were then genotyped using the matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) method in 332 buffaloes. Two of eight SNPs were not in Hardy-Weinberg equilibrium. Based on the SNP data, seven haplotypes were constructed. Three SNPs H1 (AGT), H2 (GAT), and H3 (GAC) accounted for 93.0% of the total individuals. Statistical analysis indicated that the SNP g.1104C>T was significantly associated with milk yield, protein, and fat percentage (P MC4R gene are associated with milk production traits and might be potential biomarkers for water buffalo breeding.

Genet. Mol. Res. 15(2): gmr8153
DOI: 10.4238/gmr.15028153
Research Article

As a proven tool, DNA barcoding can identify species rapidly and unambiguously. In this study, we used mtDNA cyt b, COI, and 16s rRNA sequences of six species of Pseudohynobius, Protohynobius puxiongensis, Liua shihi, Ranodon sibiricus, and Pachyhynobius shangchengensis, to reconstruct the phylogenetic relationships using Bayesian inference and maximum likelihood methods. Approximate lineage divergence times were also estimated, the divergence between them was calculated to have taken place mainly in Miocene. Our results showed that: 1) Ps. guizhouensis is an independent and valid species that is a sister species to Ps. kuankuoshuiensis; 2) five Pseudohynobius species formed a monophyletic group; 3) Ps. tsinpaensis is different from L. shihi, and should be classified as belonging to the Liua genus; and 4) Pr. puxiongensis is the sister lineage to all Pseudohynobius species, and should therefore be named Pseudohynobius puxiongensis.

Genet. Mol. Res. 15(2): gmr8155
DOI: 10.4238/gmr.15028155
Research Article

Feed efficiency and carcass characteristics are late-measured traits. The detection of molecular markers associated with them can help breeding programs to select animals early in life, and to predict breeding values with high accuracy. The objective of this study was to identify polymorphisms in the functional and positional candidate gene NEUROD1 (neurogenic differentiation 1), and investigate their associations with production traits in reference families of Nelore cattle. A total of 585 steers were used, from 34 sires chosen to represent the variability of this breed. By sequencing 14 animals with extreme residual feed intake (RFI) values, seven single nucleotide polymorphisms (SNPs) in NEUROD1 were identified. The investigation of marker effects on the target traits RFI, backfat thickness (BFT), ribeye area (REA), average body weight (ABW), and metabolic body weight (MBW) was performed with a mixed model using the restricted maximum likelihood method. SNP1062, which changes cytosine for guanine, had no significant association with RFI or REA. However, we found an additive effect on ABW (P ≤ 0.05) and MBW (P ≤ 0.05), with an estimated allele substitution effect of -1.59 and -0.93 kg0.75, respectively. A dominant effect of this SNP for BFT was also found (P ≤ 0.010). Our results are the first that identify NEUROD1 as a candidate that affects BFT, ABW, and MBW. Once confirmed, the inclusion of this SNP in dense panels may improve the accuracy of genomic selection for these traits in Nelore beef cattle as this SNP is not currently represented on SNP chips.

Genet. Mol. Res. 15(2): gmr8161
DOI: 10.4238/gmr.15028161
Research Article

We investigated the effect of the IL-17 monoclonal antibody Secukinumab combined with IL-35 in the blockade of the Notch signaling pathway on the invasive capability of hepatoma cells. We examined the effects of IL-17 antibody or IL-35 treatment alone or in combination on cell invasion and migration capabilities with Transwell chambers. The mRNA levels of Hes1, Hes5, and Hey1 were tested using quantitative polymerase chain reaction. The protein expression of N1ICD, Snail, and E-cadherin protein expressions were measured with western blot. The expression of Hes1, Hes5, Hey1 and N1ICD were all very high in hepatoma cell lines, and were positively correlated with the invasive migration capabilities of the cells. The combination of IL-17 monoclonal antibody Secukinumab with IL-35 could effectively inhibit the Notch signaling pathway, as well as the invasive migration of the cells. Snail and E-cadherin are involved in the migration of hepatoma cells, and it has been established that Snail can regulate the expression of E-cadherin. IL-17 monoclonal antibody Secukinumab combined with IL-35 can increase E-cadherin and decrease Snail expression, which are positively correlated with cell invasive migration capabilities. Overall, treatment with both IL-17 antibody and IL-35 is more effective than each treatment alone. Notch signaling is activated in hepatoma cell lines and increases with the enhancement of cell invasive migration capabilities. IL-17 monoclonal antibody Secukinumab combined with IL-35 can block the Notch signaling pathway, simultaneously reducing the invasive migration capability of hepatoma cells.

Genet. Mol. Res. 15(2): gmr8174
DOI: 10.4238/gmr.15028174
Research Article

We investigated the effects of eribulin and paclitaxel on breast cancer (BC) by exploring molecular biomarkers and pathways. Co-expression networks were constructed by differentially co-expressed genes and links, and centralities were analyzed to explore the hub genes. Pathway-enrichment analysis was performed. The hub genes were validated using the polymerase chain reaction and western blotting. A total of 132 and 153 differentially expressed genes were identified in BC cell lines treated with eribulin and paclitaxel, respectively. Six hub genes were identified in two co-expression networks. The spliceosome pathway was the mutually significant pathway. The validation analysis was basically consistent with the bioinformatics. We successfully identified several hub genes and pathways relevant to the effects of eribulin and paclitaxel on BC based on the network analysis.

Genet. Mol. Res. 15(2): gmr8192
DOI: 10.4238/gmr.15028192
Research Article

The aim of this study was to evaluate the effects of miR-26a on Beclin 1 expression in retinoblastoma (RB) cell lines (Y79 and WERi-RB-1). RB cells were transfected with miR-26a mimic, antagomir-26a, or control mimic. The Beclin 1 mRNA and protein levels were detected by quantitative polymerase chain reaction and western blot, respectively. The activity of Beclin 1 3ꞌ-UTR reporter gene was detected with the luciferase assay. After transfection with miR-26a mimic, Beclin 1 mRNA and protein levels as well as the activity of the 3'-UTR reporter gene decreased. However, all were increased upon inhibition of miR-26a with antagomir-26a. Beclin 1 is the target of miR-26a in human RB cell lines Y79 and WERi-RB-1, and miR-26a inhibits the expression of Beclin 1 by reducing its mRNA and protein levels.

Genet. Mol. Res. 15(2): gmr8193
DOI: 10.4238/gmr.15028193
Research Article

The objective of this study was to examine the association between TNF-α serum levels and -308G>A and -238G>A polymorphisms in the corresponding gene by comparing healthy subjects to colorectal cancer (CRC) patients from a Mexican population. Serum levels of TNF-α were found to significantly differ between CRC patients and controls (P = 0.001), but no relationship between the -308G>A and -238G>A polymorphisms and increased CRC risk was established (P > 0.05). However, an association between the -308G>A variant and disease became evident when the distribution of AA-GA genotypes was examined in patients with hematologic toxicity (neutropenia) and those without (odds ratio = 3.356, 95% confidence interval = 1.295- 8.698, P = 0.013). The GG haplotype was more common in controls than CRC patients, with a frequency of 0.85 among the former, but this difference was not significant (P > 0.05). In conclusion, TNF-α serum levels and AA-AG genotypes of the TNF-α-308G>A polymorphism may significantly contribute to CRC susceptibility in the population examined in this investigation.

Genet. Mol. Res. 15(2): gmr8199
DOI: 10.4238/gmr.15028199
Research Article

In this study, we sequenced the complete mitochondrial DNA of Chinese indigenous Jinhu Black-bone and Rugao chickens. The two chicken mitochondrial genomes were deposited in GenBank under accession Nos. KP742951 and KR347464, respectively. The complete mitochondrial genomes of Jinhu Black-bone and Rugao chickens were sequenced and found to span 16,785 and 16,786 bp, respectively, and consisted of 22 tRNA genes, two rRNA genes (12S rRNA and 16S rRNA), 13 protein-coding genes, and one control region (D-loop). The majority of genes were positioned on the H-strand, and the ND6 and eight tRNA genes were found to be encoded on the L-strand. The mitogenomes showed a similar gene order to that of the published Gallus gallus genome, as neither included a control region. The overall base composition of the genome of the two chickens was A = 30.22/30.28%, G = 13.57/13.49%, T = 23.74/23.76%, and C = 32.48/32.48%. Nucleotide skewness of the coding strands of the two chicken genomes (AT-skew = 0.12, GC-skew = -0.41) was biased towards T and G. Phylogenetic analysis revealed 29 subspecies, and the molecular genetic relationship among the 29 subspecies was identical to that of traditional taxonomy.

Genet. Mol. Res. 15(2): gmr8200
DOI: 10.4238/gmr.15028200
Research Article

We investigated the association between serum visfatin levels and single nucleotide polymorphisms (SNPs; rs61330082, rs2058539) in the visfatin gene and coronary artery calcification (CAC) in patients from Wenzhou, China. CAC patients (N = 206) were divided into two groups: mild CAC (MCAC) and moderate and severe CAC (MSCAC). Volunteers without CAC (N = 70) were included in the control group. The serum visfatin level was analyzed by enzyme-linked immunosorbent assay. SNPs (rs61330082, rs2058539) in the visfatin gene were analyzed by polymerase chain reaction-restriction fragment length polymorphism. Clinical data, serum visfatin levels, and genotype and allele frequencies of rs61330082 and rs2058539 were compared among the three groups. MSCAC patients expressed significantly higher serum visfatin levels (30.58 ± 6.12 ng/mL) than individuals in the MCAC (29.03 ± 1.87 ng/mL) and control (24.45 ± 5.44 ng/mL) groups (P

Genet. Mol. Res. 15(2): gmr8217
DOI: 10.4238/gmr.15028217
Research Article

ATP citrate lyase (ACLY) is the primary enzyme responsible for the synthesis of cytosolic acetyl-CoA, which is a key precursor of both fatty acid and mevalonate synthesis pathways. Genetic variation of the ACLY gene may influence multiple traits associated with animal production. Here, we identified three non-synonymous mutations in ACLY exons in five beef cattle populations using DNA pool sequencing and high-resolution melting analysis. Results from association analyses revealed that the single nucleotide polymorphism (SNP) g.17127C>T is significantly associated with chest girth (P C is significantly associated with an increase in chest girth (P ACLY gene are associated with growth traits in beef cattle in northwest China. However, a larger sample set is needed to validate these findings.

Genet. Mol. Res. 15(2): gmr8250
DOI: 10.4238/gmr.15028250