Role of LncRNA CASC9 in oral carcinoma and its downstream gene expression and mechanism detected by nanomagnetic bead-based polymerase chain reaction

2021 ◽  
Vol 11 (7) ◽  
pp. 1017-1023
Author(s):  
Junjie Liu ◽  
Hong Mu ◽  
Xiangyong Cheng ◽  
Gangying Yuan ◽  
Qinqin Wang ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Oral Carcinoma ◽  
Luciferase Reporter ◽  
Chain Reaction ◽  
Total Rna ◽  
Polymerase Chain ◽  
And Migration ◽  
The Impact ◽  
Downstream Gene Expression

In view of the increasing incidence of oral carcinoma (OC), an in-depth understanding of the mechanism is required for future prevention and treatment. In modern medical research, the etiology of tumor diseases is mainly focused on gene changes, among which lncRNA is a trending topic. We speculated that lncRNA CASC9 may attribute to OC occurrence. To verify our conjecture, we extracted the total RNA of the research samples through nanomagnetic beads, detected the expression of CASC9 downstream genes miR-383-5p and KIF3B by polymerase chain reaction (PCR), and analyzed the impact of the three on OC cells and the targeting relationship. The total RNA extracted by nanomagnetic beads were found to be able to effectively improve the PCR speed, yield and the accuracy of the results, and reduce the allergic amplification results of the samples to be tested to obtain the best results. The detection result showed high CASC9 and KIF3B expression, and low miR-383-5p expression in OC cells. Inhibition of CASC9 and KIF3B and elevation of miR-383-5p can inhibit the viability of OC cells and boost apoptosis. Through dual-luciferase reporter (DLR) assay, the targeted regulation relationship between CASC9 and miR-383-5p and between miR-383-5p and KIF3B, was determined. Rescue experiments confirmed that CASC9 was able to modulate OC cell biological behaviors by targeting the miR-383-5p/KIF3B axis. Therefore, we argue that with high expression in OC, CASC9 can enhance the proliferation and migration of OC cells and inhibit the apoptosis through targeting the miR-383-5p/KIF3B axis.

scholarly journals Macrophage migration inhibitory factor may play a protective role in osteoarthritis

2021 ◽  
Vol 23 (1) ◽  
Author(s):  
Ming Liu ◽  
Zikun Xie ◽  
Guang Sun ◽  
Liujun Chen ◽  
Dake Qi ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Migration Inhibitory Factor ◽  
Macrophage Migration Inhibitory Factor ◽  
Protective Role ◽  
Macrophage Migration ◽  
Chain Reaction ◽  
Protein Levels ◽  
Tnf Α ◽  
Polymerase Chain

Abstract Background Osteoarthritis (OA) is the most prevalent form of arthritis and the major cause of disability and overall diminution of quality of life in the elderly population. Currently there is no cure for OA, partly due to the large gaps in our understanding of its underlying molecular and cellular mechanisms. Macrophage migration inhibitory factor (MIF) is a procytokine that mediates pleiotropic inflammatory effects in inflammatory diseases such as rheumatoid arthritis (RA) and ankylosing spondylitis (AS). However, data on the role of MIF in OA is limited with conflicting results. We undertook this study to investigate the role of MIF in OA by examining MIF genotype, mRNA expression, and protein levels in the Newfoundland Osteoarthritis Study. Methods One hundred nineteen end-stage knee/hip OA patients, 16 RA patients, and 113 healthy controls were included in the study. Two polymorphisms in the MIF gene, rs755622, and -794 CATT5-8, were genotyped using polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) and PCR followed by automated capillary electrophoresis, respectively. MIF mRNA levels in articular cartilage and subchondral bone were measured by quantitative polymerase chain reaction. Plasma concentrations of MIF, tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and interleukin-1 beta (IL-1β) were measured by enzyme-linked immunosorbent assay. Results rs755622 and -794 CATT5-8 genotypes were not associated with MIF mRNA or protein levels or OA (all p ≥ 0.19). MIF mRNA level in cartilage was lower in OA patients than in controls (p = 0.028) and RA patients (p = 0.004), while the levels in bone were comparable between OA patients and controls (p = 0.165). MIF protein level in plasma was lower in OA patients than in controls (p = 3.01 × 10−10), while the levels of TNF-α, IL-6 and IL-1β in plasma were all significantly higher in OA patients than in controls (all p ≤ 0.0007). Multivariable logistic regression showed lower MIF and higher IL-1β protein levels in plasma were independently associated with OA (OR per SD increase = 0.10 and 8.08; 95% CI = 0.04–0.19 and 4.42–16.82, respectively), but TNF-α and IL-6 became non-significant. Conclusions Reduced MIF mRNA and protein expression in OA patients suggested MIF might have a protective role in OA and could serve as a biomarker to differentiate OA from other joint disorders.

scholarly journals Clinical assessement of the 8 genes on chromosome 3 with also MGMT gene promoter regions methylaton status in patients with breast cancer luminal hystotype

2017 ◽  
Vol 16 (4) ◽  
pp. 38-45
Author(s):  
D. A. Ryabchikov ◽  
I. K. Vorotnikov ◽  
T. P. Kazubskaya ◽  
S. S. Lukina ◽  
E. A. Filippova ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Polymerase Chain Reaction ◽  
Normal Tissue ◽  
Methylation Status ◽  
Chromosome 3 ◽  
Epigenetic Changes ◽  
Chain Reaction ◽  
Promoter Regions ◽  
Polymerase Chain

Background. Epigenetic changes of TSG are supposed as the most fine and active genes regulation mechanism in particular breast cancer (BC) genes pathway development. The most valuable results are awaited for methylation role of genes located on the short arm of chromosome 3 with also MGMT gene (10q26) in BC pathogenesis because of their ambiguous data for methylation status in tumors. Objective: to illustrate the specific methylation role of the RASSF1A, SEMA3B, RARß2, RHOA, GPX1, USP4, DAG1, NKIRAS1 and MGMT genes promoter regions in BC pathogenesis. Materials and methods. Sample set of 174 BC patients consists of tumor and surrounding histologically normal tissue that were collected and clinically characterized in the N.N. Blokhin National Medical Research Center of Oncology. Two substantive methods were used to evaluate DNA methylation status. To analyse RASSF1A, SEMA3B, RARß2 and MGMT genes methylation we used polymerase chain reaction specific for the methylated allele. Whereas for analyses RHOA, GPX1, USP4, DAG1, NKIRAS1 promoter regions genes methylation status was used methyl sensitive restriction analyses with 2 methyl sensitive endonuclaeses HpaII and HhaI with subsequent polymerase chain reaction. Results. A statistically significant high frequency of RASSF1A, SEMA3B, RARß2, and MGMT genes methylation in epithelial breast tumors compared with histologically normal tissue from the same patients was shown. Significant correlation of RARß2 and MGMT genes methylation frequency considering the different clinical and morphological characteristics of the malignant process was revealed. The statistically significant relationship between methylation of RASSF1A, RARß2 and MGMT genes and patient survival is shown for the first time. Conclusion. The findings of epigenetic changes in the luminal BC supplement the “molecular picture” of this cancer and contribute to an understanding of its pathogenesis. The revealed features of investigated genes methylation can find clinical application for the development of modern approaches to prognosis, prevention and choice of tactics for treatment of BC in females of the Moscow region.

Detection of Aspergillus fumigatus by Polymerase Chain Reaction (PCR)

2019 ◽  
Vol 10 (04) ◽  
pp. 655-661
Author(s):  
Zainab H Abood AL-Asadi
Keyword(s):  
Polymerase Chain Reaction ◽  
Aspergillus Fumigatus ◽  
Oligonucleotide Primer ◽  
Rrna Gene ◽  
Fungal Culture ◽  
Chain Reaction ◽  
Isolation And Identification ◽  
Internal Transcribed Spacer Region ◽  
Polymerase Chain ◽  
The Impact

Aspergillosis refers to fungi infections of the respiratory tract caused by Aspergillus species, especially Aspergillus fumigatus. Infection of A. fumigatus was increased in the last few years due to either resistances to antibiotics or the influence of other factors such as other fungal infections. The present study aimed to review the impact of Aspergillus fumigatus in Aspergillosis cases, and study the role of Singleplex PCR for amplification of ITS1, ITS4 of rRNA gene in the detection of fungal isolate. In this study, One hundred sputum samples were collected from patients admitted to the specialize chest and respiratory diseases center / Baghdad who were suffering from respiratory problems. During these studied, molds were isolation and identification based on Conventional method (Direct microscopy by using 10% KOH, and fungal culture was done on Sabouraud Dextrose agar supplemented with chloramphenicol and on Czapek-Dox agar incubated at 37°C and examined for 3-7 days then macroscopic, microscopic examination of the colony by(lactophenol cotton blue stain )and molecular methods by using Polymerase chain reaction (PCR)technique for identification. The 10% KOH examination was positive for 35 cases, while laboratory culturing was positive for 53 cases. Aspergillus sp were isolated from 44(83%) patients; A. fumigatus was isolated in 23 (42. 4%) patients while A. flavus, A. niger, and A. terreus were isolated from 11 (20. 08%), (13. 2%) and 3 (5. 7%) patients respectively, also isolated Penicillium spp. at percentage 1(1. 9%). In this study. The ages of participants ranged from 10-70years with a mean age of 34years, the males were more susceptible to fungal infection, were recorded 35/53 (66. 3), compared to females were 18/53 (33. 96). The infection of fungi was more prevalent in ages 30-40recorded 26(53. 06%) followed by ages 40-50, 13(26. 5), while the lowest infection recorded in the age group 10- 20 years was 2(2. 04%). DNA isolated from twenty-three A. fumigatus isolates was used as a template, and the specific of oligonucleotide primer sequences were used in conventional PCR to detect the presence of internal transcribed spacer region ( ITS) region of the rRNA gene for Aspergillus fumigates. The results of the PCR amplification of the rRNA gene showed that this gene was present in 19 samples out 23 positive samples which isolation with a PCR product size of approximated 385 bp, while 4 samples out 23 positive samples showed negative results for the presence of this gene as indicated by the absence of the PCR products in their relevant lanes. Statistical analysis revealed that the PCR to have a sensitivity of 95. 1% in the detection of Aspergillus fumigatus in Aspergillosis cases. Polymerase chain reaction (PCR) is a rapid, specific, and sensitive method to detect Aspergillus fumigatus in aspergillosis cases of humans.

The role of intraocular f luid polymerase chain reaction in the diagnosis, treatment and monitoring of cytomegalovirus retinitis

2021 ◽  
pp. 380-383
Author(s):  
B.S. Pershin ◽  
◽  
A.A. Maschan ◽  
V.Y. Makhmutov ◽  
M.A. Ilushina ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Stem Cell ◽  
Retinal Detachment ◽  
Cytomegalovirus Retinitis ◽  
Chain Reaction ◽  
Hematopoietic Stem ◽  
Key Words Cytomegalovirus ◽  
Polymerase Chain ◽  
Intraocular Fluid

Purpose. To study the possibilities of a new method of CMRR treatment in the prevention of irreversible blindness. Material and methods. 74 patients with cytomegalovirus retinitis, frolicking after hematopoietic stem cell transplantation. The first group (9 people, 15 eyes) consisted of children, whose treatment was carried out under ophthalmoscopic control. The second group (65 people, 114 eyes) consisted of children in whom the control of the effectiveness of treatment was carried out using PCR of aqueous humor in real time. Results. In the first group, retinal detachment was diagnosed in three out of fifteen eyes, accounting for 20%. In the second group, the incidence of retinal detachment was 3.5% of 114 eyes. Among patients receiving treatment under ophthalmoscopic control, CMRR relapses were detected in 5 cases, which amounted to 33.3%. In children, whose treatment was controlled by intraocular fluid PCR, relapses were diagnosed in 22 cases, which amounted to 19.29%. Conclusions. Intravitreal administration of antiviral drugs under the control of polymerase chain reaction is a more effective method of treating cytomegalovirus retinitis than intravitreal administration under ophthalmoscopic control. Key words: cytomegalovirus retinitis, intraocular fluid polymerase chain reaction.

scholarly journals The impact of COL1A1 and COL6A1 expression on hypospadias and penile curvature severity

BMC Urology ◽  
2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Prahara Yuri ◽  
Gunadi ◽  
Rahmadani Puji Lestari ◽  
Firly Putri Fardilla ◽  
Wiwit Ananda Wahyu Setyaningsih ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Polymerase Chain Reaction ◽  
External Genitalia ◽  
Penile Curvature ◽  
Moderate Correlation ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Distal Hypospadias ◽  
Male External Genitalia ◽  
The Impact

Abstract Background Hypospadias, the most frequent congenital male external genitalia abnormality, is usually associated with curvature of the ventral penis, i.e. chordee. Abnormality of darto tissue has been suggested as the pathophysiology of chordees. Collagen is one of the most abundant fibrous proteins within the extracellular matrix. In this study, we determined the expression of collagen 1 (COL1A1) and COL6A1 in patients with hypospadias and associated them with the severity of penile curvature. Methods We included 60 children < 18 years old, consisting of 20 distal hypospadias, 20 proximal hypospadias patients, and 20 controls in our institution from 2017 – 2020. The expression of COL1A1 and COL6A1 in darto tissue was determined by reverse-transcriptase polymerase chain reaction (qPCR). The penile curvature severity was classified as mild (< 30 degrees), moderate (30–60 degrees), and severe (> 60 degrees). Results qPCR showed that COL1A1 and COL6A1 expression was significantly downregulated in the distal (0.88 (0.38–2.53) and 0.54 (0.16–4.35), respectively) and proximal 0.76 (0.33–2.57) and 0.57 (0.18–1.38), respectively) hypospadias groups compared to controls (1.85 (0.24–4.61) and 0.93 (0.17–4.06), respectively) with p-values of 0.024 and 0.018, respectively. Furthermore, there was a moderate correlation between COL1A1 and COL6A1 expression (r = 0.458, p < 0.0001). Interestingly, COL1A1 and COL6A1 were also significantly downregulated in the moderate and severe chordee groups compared to the mild chordee groups, with p-values of 0.003 and 0.037, respectively. Conclusions Aberrant COL1A1 and COL6A1 expression might affect abnormalities in darto tissue and penile curvature severity in hypospadias patients.

Study of the HLA-DPβ1 Locus by the Polymerase Chain Reaction Technique in Patients with Hodgkin's Disease

1993 ◽  
Vol 79 (2) ◽  
pp. 133-136 ◽  
Author(s):  
Guseppe Pellegris ◽  
Claudia Lombardo ◽  
Annelisa Cantoni ◽  
Liliana Devizzi ◽  
Monica Balzarotti
Keyword(s):  
Polymerase Chain Reaction ◽  
Hodgkin's Disease ◽  
Hodgkin’S Disease ◽  
Polymerase Chain Reaction Method ◽  
Healthy Controls ◽  
Chain Reaction ◽  
Reaction Method ◽  
Significant Difference ◽  
Polymerase Chain

Background A number of reports have studied associations between Hodgkin's disease and HLA. Some of them established correlation between several antigens and Hodgkin's disease, and others found no correlations. Methods The HLA DP locus was determined by the polymerase chain reaction method in 31 Hodgkin's disease patients and 58 healthy controls. Results No significant difference between patients and controls was noted. Conclusions Further investigations are needed to confirm the hypothesis of a possible role of the HLA complex as one of the factors involved in Hodgkin's disease.

Sign in / Sign up

Export Citation Format

Share Document