The Journal of Gene Medicine最新文献

Background: As a result of the growing prevalence of colorectal cancer (CRC), new screening and early detection methods are required. Among the novel biomarkers, DNA methylation has emerged as a high-potential diagnosis/screening molecular marker. The present study aimed to assess non-invasive early diagnosis of CRC by examining promoter methylation of TFPI2 and NDRG4 genes in peripheral blood mononuclear cells (PBMCs).

Methods: Fifty CRC patients and 50 normal controls were recruited to the present study. Quantitative methylation of the promoter region of the TFPI2 and NDRG4 genes was analyzed in DNA extracted from PBMCs of all cases and control subjects using a methylation-quantification endonuclease-resistant DNA (MethyQESD) method.

Results: The sensitivity and specificity of the TFPI2 gene for the diagnosis of CRC was 88% and 92%, respectively, and, for the NDRG4 gene, it was 86% and 92%, respectively. The methylation range for the TFPI2 gene was 43.93% and 11.56% in patients and controls, respectively, and, for the NDRG4 gene, it was 38.8% in CRC patients and 12.23% in healthy controls (p < 0.001). In addition, we observed that a higher percentage of methylation was correlated with the higher stage of CRC.

Conclusions: The results of the present study reveal that PBMCs are reliable sources of methylation analysis for CRC screening. Furthermore, the TFPI2 and NDRG4 genes provide sufficiently high sensitivity and specificity to be nominated for use in a novel noninvasive CRC screening method in PBMCs.

Background: The present study aimed to investigate the effect of cnidium lactone on ovariectomy (OVX)-induced bone loss and determine whether it exerts its effects by mediating the estrogen receptor-α (ERα)/bone morphogenetic protein-2 (BMP-2)/Smad signaling pathways.

Methods: Fifty-five female rats were randomly assigned to the following treatment groups: the OVX group, the sham-operated (sham) group, and groups treated with cnidium lactone at different doses (10 mg/kg/day, 20 mg/kg/day, 30 mg/kg/day). Treatments were administered for 60 days. Search Tool for Interacting Chemicals (STITCH; http://stitch.embl.de) was used to identify the interaction between cnidium lactone and target proteins. Bone mineral density (BMD), mechanical strength, serum osteoblastic and osteoclastic markers, and hematoxylin and eosin (HE) staining of the distal femur were evaluated. Moreover, western blot analyses were also performed to evaluate the effect of cnidium lactone on the ERα/BMP-2/Smad signaling pathway.

Results: Cnidium lactone treatment was associated with an increase in the BMD of the distal femur compared to that of the OVX group. Moreover, cnidium lactone significantly increased biomechanical properties in a dose-dependent manner compared to those of the OVX group (p < 0.05). Treatment with cnidium lactone significantly enhanced the BMP-2/Smad signaling pathway by up-regulating the expression of ERα, BMP-2, p-Smad1 and p-Smad4. Cnidium lactone treatment improved the microstructure of trabecular bone in the distal femurs of OVX rats, as shown by HE staining.

Conclusions: Cnidium lactone exerts potent antiosteoporotic activity in ovariectomized mice, and the underlying molecular mechanism may be related to the ERα/BMP-2/Smad signaling pathways.

Background: Progressive spastic ataxia is a heterogeneous disorder characterized by cerebellar ataxia and limb spasticity associated with other severe neurological complications. Spastic ataxia is classified into pure and complex types, inherited in both an autosomal recessive and autosomal dominant manner. It is caused by pathogenic variants in at least eight different genes, including NKX6-2 (MIM 607063) located on chromosome 10q26.3. The present study aimed to identify the genetic variant(s) underlying progressive spastic ataxia and to establish the genotype-phenotype correlation.

Methods: We collected a large consanguineous family having four affected individuals segregating progressive spastic ataxia in an autosomal recessive manner. To investigate the molecular cause of the disease, genomic DNA of three affected individuals underwent whole exome sequencing.

Results: All of the affected individuals showed progressive clinical features such as spastic ataxia, lower limb weakness and other mild neurological abnormalities. Whole exome sequencing data were analyzed using different filters. Filtering of rare and shared homozygous variants revealed a novel homozygous missense variant (c.545C>T; p.Ala182Val) in a highly conserved homeobox domain of the NKX6-2 protein.

Conclusions: The findings of the present study add a novel variant to the NKX6-2 mutation spectrum and provide evidence that homozygous variants in the NKX6-2 cause progressive spastic ataxia associated with other abnormalities.

Background: Sitosterolemia (STSL), also known as phytosterolemia, is a rare autosomal recessive hereditary disease caused by mutations in the ABCG5 or ABCG8 genes. The disease is a result of disorders in lipoprotein metabolism, and is characterized by tendinous and tuberous xanthomas, elevated plasma cholesterol and phytosterol levels, and thrombocytopenia and hemolytic anemia in several patients. The manifestations of STSL are diverse and can easily be misdiagnosed. In recent years, cases of this disease in children have been reported in succession. There is therefore a need for clinicians to improve identification of STSL and perform early intervention.

Methods: We evaluated four children with STSL caused by genetic mutations in ABCG5 or ABCG8, as well as their family members, by analyzing their clinical characteristics and performing Trio-whole exome sequencing. The biological consequences of the mutations were analyzed using various bioinformatics software. We also analyzed the consequences of a mutation commonly observed in STSL patients on the structure of the protein involved.

Results: We identified five previously unreported pathogenic mutations of different phenotypes of STSL: ABCG5 NM_022436:c.1337G>A; ABCG8 NM_022437:c.965-1G>A, c.323-1G>C, c.1418C>G and c.1534G>A. We also report the structural changes brought about by a mutation common in STSL patients, as well as the possible consequences of these changes.

Conclusions: Our findings further broaden the genotypic and phenotypic profiles of the onset of STSL in the pediatric population and provide information for the diagnosis and treatment of this disease.

Background: As a potential anti-arthritic agent, Andrographolide (And) is capable of promoting chondrocyte proliferation and preventing apoptosis in pathologic condition. The present study aimed to explore the roles of And in in vivo and in vitro models of osteoarthritis (OA), as well as its underlying molecular mechanisms.

Methods: An OA mouse model was established using anterior cruciate ligament transection operation on the left knee joint. The pathological changes of articular cartilage were assessed using safranin O staining. Chondrocyte proliferation and apoptosis were measured using cell a counting kit-8 assay and flow cytometry. Bioinformatics algorithms and a luciferase reporter assay were used to evaluate matrix metalloproteinase13 (MMP13) as a direct target of miR-27-3p.

Results: And had the ability to prevent catabolism and facilitate anabolism of articular cartilage in an experimental OA model in mice. In addition, And alleviated chondrocyte apoptosis in in vitro and in vivo models of OA. We also found that both up-regulation of MMP13 and down-regulation of miR-27-3p in the proximal tibia of OA mice and interleukin (IL)-1β-stimulated chondrocytes were reversed by And administration simultaneously. MMP13 was validated as direct target of miR-27-3p and could be suppressed by overexpression of miR-27-3p in mouse chondrocyte. Furthermore, overexpression of miR-27-3p or MMP13 loss-of-function in chondrocytes could alleviate IL-1β-induced apoptosis.

Conclusions: These results indicated that miR-27-3p/MMP13 signaling axis might be a potential therapeutic target of And for preventing the progression of OA.

Background: Autophagy is closely associated with apoptosis in H9c2 cardiomyocytes as a result of hypoxia. The present study aimed to determine whether microRNAs (miRs) mediated apoptosis and autophagy in hypoxia-stimulated H9c2 cardiomyocytes.

Methods: miR microarrays and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assays were used to detect differentially expressed miRs in H9c2 cardiomyocytes following hypoxia stimulation. Annexin V-fluorescein isothiocyanate double staining was performed to evaluate hypoxia-induced cell apoptosis, and the protein expression levels of autophagy-associated genes were detected using western blotting.

Results: miR microarrays and qRT-PCR assays showed that the expression of miR-129-5p is significantly decreased in hypoxia-exposed H9c2 cardiomyocytes. Under hypoxic stimulation, miR-129-5p mimics alleviated hypoxia-induced cell apoptosis and also restored autophagy in H9c2 cardiomyocytes. However, transfection with miR-129-5p inhibitors accelerated hypoxia-induced cell apoptosis and autophagy deficiency in H9c2 cardiomyocytes. Furthermore, overexpression of miR-129-5p enhanced cell viability and reduced the release of lactate dehydrogenase in hypoxia-stimulated H9c2 cardiomyocytes.

Conclusions: These findings highlight the protective effect of miR-129-5p against hypoxia-induced apoptosis in H9c2 cardiomyocytes through the activation of autophagy.

Background: The present study aimed to determine the accuracy (Z-value) of non-invasive prenatal testing (NIPT) results for sex chromosome aneuploidy (SCA) in routine clinical practice.

Methods: Among a cohort of 12505 pregnant females, maternal plasma samples collected from our hospital were utilized for SCA analysis by NIPT detection. The positive samples were validated through an invasive procedure and karyotyping analysis. The predictive value from positive samples in sex chromosomes was compared to analyze the accuracy of the Z-value.

Results: There were 65 females with sex chromosome abnormalities within 12,505 pregnant females in the NIPT detection, which was validated by karyotype analysis of amniotic fluid puncture through sequencing, as well as bioinformatics analysis, with 18 true-positive samples. The true-positive results with 45,X, 47,XXY, 47,XXX and 47,XYY karyotypes predicted by NIPT were 14.29%, 50.00%, 66.67% and 71.43%, respectively. Among sex chromosome cases, the findings indicated that positive NIPT results with Z ≥ 9 show a higher accuracy.

Conclusions: The findings of the present study demonstrate that the positive predictive value of NIPT for sex chromosome abnormalities is distinctive. The positive predictive value was highest for 47,XYY and lowest for 45,X. Additionally, the Z-value results are considered to be correlated with the accuracy of NIPT, although further studies need to be made.

Background: Breast cancer is the leading cause of cancer deaths in women worldwide. The purpose of the current study was to investigate the potential role of miR-96-5p in breast cancer.

Methods: Breast cancer tissues and matched para-cancerous tissues were collected. The expression of microRNA-96-5p (miR-96-5p) and arginine kinase 3 (AK3) was detected by quantitative real-time PCR (qRT-PCR). The correlation between miR-96-5p and AK3 was calculated by Pearson's Chi-square test. Moreover, mimics or inhibitors of miR-96-5p were applied to explore whether miR-96-5p influences the migration capacity in Transwell and wound healing assays. Bioinformatics analysis was performed to identify the target genes of miR-96-5p through the TargetScan, miRDB and miRanda databases. A luciferase reporter assay was performed to verify AK3 as a downstream target gene of miR-96-5p.

Results: The expression of miR-96-5p was significantly increased in breast cancer tissue and breast cancer cell lines compared with para-cancerous tissue and a breast cell line, respectively. The expression of miR-96-5p negatively correlated with AK3 gene expression. AK3 was demonstrated to be a direct mRNA target of miR-96-5p. AK3 was positively associated with the overall survival of breast cancer patients. Kaplan-Meier curve and log rank test analyses revealed that decreased AK3 levels were significantly associated with reduced overall survival. miR-96-5p was shown to promote the migration of breast cancer cells through the MEK/ERK signaling pathway.

Conclusion: Our results identify a role for miR-96-5p in promoting breast cancer cell migration through activation of MEK/ERK signaling by targeting AK3.

Background: Prior studies have noted the importance of T cell immunoglobulin and mucin domain containing 4 (TIMD4) in various diseases and its functions on cell malignant behaviors. However, the biological function of TIMD4 in diffuse large B-cell lymphoma (DLBCL) is unknown.

Methods: Relative expression of TIMD4 was analyzed based on the GSE56315 array including 88 cases of human tissues. TIMD4 expression in cells was detected using a quantitative reverse transcriptase-polymerase chain reaction and western blot experiments. Cell proliferation was measured using the cell counting kit-8 (CCK-8) assay and apoptotic properties were assessed through the detection of related proteins by western blotting. The underlying molecular mechanism of TIMD4 in DLBCL was predicted and confirmed using KEGG enrichment analysis and western blotting.

Results: The results indicate that TIMD4 is overexpressed in DLBCL tissues and the poor prognosis of DLBCL patients is significantly linked with the higher TIMD4 expression. The loss-of-TIMD4 experiment in CYP6D reveals that knockdown of TIMD4 blocks cell growth and accelerates cell apoptosis, whereas the gain-of-TIMD4 experiment in Raji cells suggests that up-regulation of TIMD4 promotes cell proliferation and inhibits cell apoptosis. The activity of the Wnt/β-catenin pathway is mediated by the TIMD4 expression in DLBCL cells.

Conclusions: These findings demonstrate that TIMD4 is up-regulated in patients with DLBCL and the regulatory effects of TIMD4 on cell proliferation and apoptosis are associated with the Wnt/β-catenin pathway, posing a novel target for DLBCL therapy.

Background: Neuroblastoma is one of the most common malignant tumors in childhood. Polymorphisms in proto-oncogene MYC are implicated in many cancers, although their role in neuroblastoma remains unclear. In the present study, we attempted to investigate the association between MYC gene polymorphisms and neuroblastoma susceptibility in Chinese children.

Methods: We included two MYC polymorphisms (rs4645943 and rs2070583) and assessed their effects on neuroblastoma risk in 505 cases and 1070 controls via the Taqman method.

Results: In single and combined locus analysis, no significant association was found between the two selected polymorphisms and neuroblastoma susceptibility. In stratification analysis, the rs4645943 CT/TT genotypes were significantly associated with a decreased neuroblastoma risk in subjects with tumors originating from other sites [adjusted odds ratio (OR) = 0.42, 95% confidence interval (CI) = 0.21-0.84, p = 0.013]. Meanwhile, the presence of one or two protective genotypes was significantly associated with a decreased neuroblastoma risk in subjects with tumors arising from other sites (adjusted OR = 0.50, 95% CI = 0.26-0.96, p = 0.036).

Conclusions: The present study indicates that MYC gene polymorphisms may have a weak effect on the neuroblastoma risk, which neeeds to be verified further.