scholarly journals Determination of expression profile of p53 gene in different grades of breast cancer tissues by real time PCR

2020 ◽  
Vol 20 (3) ◽  
pp. 1273-1282
Author(s):  
Haleema Sadia ◽  
Munir Ahmad Bhinder ◽  
Asma Irshad ◽  
Beenish Zahid ◽  
Rais Ahmed ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Tumor Suppressor ◽  
Real Time ◽  
Real Time Pcr ◽  
P53 Gene ◽  
Abnormal Behavior ◽  
Internal Stresses ◽  
Down Regulation ◽  
Significant Difference

Background: Pakistan has a high incidence of breast cancer in Asia, where annually 16,232 deaths are reported. There are many exogenous and endogenous risk factors that affect the tumor suppressor genes and oncogenes. The p53 gene is a tumor suppressor gene and it has a role to protect the whole genome from external and internal stresses, which causes damages to the genome. Objective: The aim of the current study was to investigate the p53 gene expression by using the real-time PCR technique in different grades of breast cancer as compared to the normal tissue. Methods: Fresh Modified Radical Mastectomy (MRM) samples (grade1-grade3) were collected from different hospitals of the Lahore. The project was approved by an ethical review committee of Jinnah Hospital, Lahore. And before sampling an informed consent was obtained from patients and clinicians. RNA from fresh biopsies was extracted by Qiagen extraction kit and cDNA was formed. Real time PCR performed by using SYBR green master mix (ABI) and the data was evaluated by using Livak method. Statistical analysis was done by using Microsoft Excel. Results: There was an abnormal gene expression of p53 in all grades of the breast tumors. Non-significant (p>0.05) differ- ence of down and up regulation of p53 in different grades of breast tumor was found. However, as a whole up-regulation was more than down-regulation with significant difference (p<0.0011). Conclusion: The abnormal expression of p53 shows that there are some genetic and epigenetic factors which are the primal cause of an abnormal gene expression. It is recommended that perform next generation sequencing (NGS) of the gene to find out the mutations causing the abnormal behavior of p53 gene. Keywords: Breast cancer; up-regulation; down-regulation; real-time PCR; Punjab; Pakistan; p53 gene.

scholarly journals P4: CIRCULATING MIRNA PROFILES POINT TO DYSREGULATION IN TGFBETA/SMAD3 TUMOR SUPPRESSOR SIGNATURE IN BRCA POSITIVE BREAST CANCER PATIENTS

2016 ◽  
Vol 64 (3) ◽  
pp. 818.2-818
Author(s):  
I Shapira ◽  
T Bhuiya ◽  
S Arora ◽  
N Mukhi ◽  
S Datla ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Tumor Suppressor ◽  
Real Time ◽  
Cancer Patients ◽  
Survival Data ◽  
Real Time Pcr ◽  
Germ Line ◽  
Germline Mutations ◽  
Breast Cancer Patients ◽  
Mean Differences

Purpose of StudyOver 240,000 individuals are diagnosed with breast cancer (BrCa) of which 12,000 individuals carry BRCA germline mutations. MicroRNA dysregulation is common in malignancy and may correlate with germline mutations.Aims:1. Analyze microRNAs in patients with breast cancer with or without BRCA germ line mutations, with and without cancer.2. Identify molecular BRCA mutant patients to deduct reasons for accelerated malignancy.Methods UsedWe analyzed plasma miR expression from 94 br cancer patients (41 BRCA positive) relative to 24 normal controls. All samples were collected between 2010 and 2014 and survival data was known for all cancer patients. TaqMan Open Array panel was used to simultaneously run hundreds of microRNA assays in the Applied Biosystem Open array real time PCR. Using AB open array real time PCR, 756 miRNA species were detected. Two-sample t-test was used for all 2-sample comparison and ANOVA followed by Tukey HSD post-hoc test to compare the miRs mean differences. All tests were 2-tailed and results with a p<0.05 were considered statistically significant.Summary of ResultsBRCA+underexpressed hsa-mir-10a and hsa-mir-376c and over-expressed Hsa- mir- 326 and Hsa-mir-143 relative to BRCA-; p<0.05.Using Coremine data mining linking genes and diseases differentially expressed circulating miRs are linked to tumor suppressor TGFbeta/SMAD3.ConclusionsThe early onset of breast cancer in BRCA mutant patients may recapitulate the pro-oncogenic effects of TGF-β. The context dependent SMAD3 binding & tumor suppression TGF-β effects are abrogated in BRCA mutant patients. TGF-β/Smad3 tumor-suppressor signature suppresses local inflammation in the tumor microenvironment.

Real-Time PCR Gene Expression Profiling of Microarray Gene Signatures in Acute Leukemia.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4401-4401
Author(s):  
Ebrahim Sakhinia ◽  
Mahboubeh Farahangpour ◽  
John A. Liu Yin ◽  
Gerard Brady ◽  
Judith A. Hoyland ◽  
...  
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Real Time Pcr ◽  
Cyclin D3 ◽  
Expression Level ◽  
Tumor Type ◽  
Gene Signatures ◽  
Significant Difference ◽  
The Mean ◽  
Microarray Gene

Abstract Cancer subtype discovery and classification using microarray gene signatures has the potential to transform pathological diagnosis but measurement of indicator genes in routine practice remains difficult. We tested use of real-time PCR measurement of indicator genes for AML and ALL (Golub et al, Science, 1999) as a method for validation and application of microarray gene signatures. Mononuclear cells (MC) were isolated from whole bone marrow (BM) aspirates by density gradient centrifugation and sorted into unselected (total), CD34+ve and CD34-ve fractions. The mRNA in each fraction was globally amplified using a PolyA PCR method. We measured the expression profile of the 17 top ranked genes (cystatin C, leptin receptor, fumarylacetoacetate, CD33, HoxA9, adipsin, proteoglycan 1, LTC4 synthase, LYN, C-myb, MB-1, cyclin D3, SNF2, RbAp48, proteasome iota, HkrT-1 and E2A) from Golub et al (1999) by real-time PCR. All values were calibrated against control standards and normalized to the mean of three housekeeping genes (IF2-beta, GAPDH and human ribosomal protein S9). Data for all 17 genes were obtained for 4 (ALL), 26 (AML), 12 (AML remission) and 9 (morphologically normal) BM samples, each fractionated into three fractions (total MC, CD34+ve MC & CD34−ve MC). There was no significant difference in the mean of three housekeeping gene expression levels between the diagnostic groups. Comparison of the expression level of the other genes confirmed ability to separate AML and ALL, whilst the direction of expression change (increased or decreased) for each gene between AML and ALL was the same as found by Golub et al. In particular, c-myb showed largest significant increase in ALL vs AML in the total BM fraction, whilst cystain c was increased in AML in the CD34−ve fraction. hSNF2b was significantly increased in the ALL total B.M fraction and Hox-A9 was significantly increased in the AML CD34+ve B.M fraction. Furthermore expression level of LYN and CD33 was significantly increased in AML compared to remission AML, indicating ability of the method to determine activity status of disease. In addition, several of the genes provided better separation between AML and ALL when measured in the CD34+ve and −ve fractions indicating more prominent expression in cells of different maturity and that prior fractionation is diagnostically more informative. The results demonstrate ability of the method to validate gene expression signatures by an independent method, which is simple, sensitive and robust, allowing translation to routine clinical use. Whilst the present study used AML and ALL, in principle the method could be extended to any other tumor type for which gene signatures exist.

scholarly journals Concurrent Down-regulation of the EAF1 and the EAF2 Genes in the Invasive Ductal Carcinoma Type of Breast Cancer

2021 ◽  
Vol 11 (2) ◽  
pp. 1620-1628
Author(s):  
Sogand Heydaran
Keyword(s):  
Breast Cancer ◽  
Tumor Suppressor ◽  
Real Time ◽  
Invasive Ductal Carcinoma ◽  
Triple Negative ◽  
Ductal Carcinoma ◽  
Rna Extraction ◽  
P Value ◽  
Control Analysis ◽  
Down Regulation

In mammals, the EAF1 and EAF2 genes build an active transcription module with other components. These genes code for factors acting as potent inhibitor of the Wnt-β catenin pathway, which might be due to their function as tumor suppressor gene. Recently, the involvement of both above-mentioned factors was described in some human tumors, but not yet in breast cancer. Breast cancer is one of the most cancer cases in Iran after colon and stomach carcinoma. We aimed therefore to investigate for the first time a possible correlation between breast cancer and the EAF1 and the EAF2 gene expression. We collected invasive ductal carcinoma tumor grading (grade 1 to 3) with marginal normal tissue from forty women diagnosed with breast cancer with the average age of 50 years old. All patients underwent triple marker test (ER/PR and Her2/neu), indeed the most of them were triple positive with few triple negative individuals. After RNA extraction, cDNA was synthesized for subsequent real-time polymerase chain reaction. The humanRPL27 gene wasusedas endogenous control. Analysis of real-time PCR results showeda significant down-regulation of EAF1 (p-value: 0.028), and of EAF2 (p-value: 0.0134) in tumor tissue samples in comparison to normal one.There was no correlation between clinical parameters and the target genes. We could find significant connection between both tumor suppressor genes in triple positive and triple negative breast cancer patients, which deserves more attention.

scholarly journals BIF-1 Relative Gene Expression Study in Breast Cancer Tumor and Normal Adjacent Tissues using Real Time PCR

2017 ◽  
Vol 25 (3) ◽  
pp. 138-146
Author(s):  
Kazhaleh Mohammadi ◽  
Mahdiyeh Salimi ◽  
Abdolhamid Angaji ◽  
Foroozandeh Mahjoobi ◽  
Tayebeh Majidizadeh ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Real Time ◽  
Real Time Pcr ◽  
Relative Gene Expression ◽  
Gene Expression Study ◽  
Expression Study ◽  
Cancer Tumor ◽  
Relative Gene

Calcium-sensitive receptor expression differs between primary and secondary hyperparathyroidism

2020 ◽  
Author(s):  
Meng Yang ◽  
Yao Lu ◽  
Ling Zhang ◽  
Aiping Song ◽  
Honglei Zhang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Primary Hyperparathyroidism ◽  
Secondary Hyperparathyroidism ◽  
Real Time ◽  
Real Time Pcr ◽  
Theoretical Basis ◽  
Receptor Expression ◽  
Down Regulation ◽  
Casr Gene ◽  
Expression Characteristics

Abstract Background: Calcium-sensitive receptor (CASR) plays an important role in the pathogenesis and progression of secondary hyperparathyroidism (SHPT). The purpose of this study is to examine the protein and gene expression characteristics of CASR in SHPT.Methods: Immunohistochemistry and real-time PCR were used to detect and compare the expression of CASR protein and genes in SHPT and primary hyperparathyroidism (PHPT) tissues.Results: CASR protein was down-regulated in SHPT and PHPT compared with normal parathyroid tissues (2.42±0.5 vs. 3.2±0.62, P<0.05; 1.8±0.83 vs. 3.2±0.62, P<0.05). Further, SHPT tissue showed higher expression of both CASR protein (2.42±0.5 vs. 1.8±0.83, P<0.05) and CASR gene (0.29±0.23 vs. 0.01±0.12, P<0.05) than PHPT tissue, respectively.Conclusion: The expression of CASR protein and gene in SHPT is higher than that in PHPT. This feature provides a theoretical basis and further ideas for studying the mechanism of CASR down-regulation.

CASR Expression Differs Between Secondary Hyperparathyroidism and Primary Hyperparathyroidism

2020 ◽  
Author(s):  
Meng Yang ◽  
Yao Lu ◽  
Ling Zhang ◽  
Aiping Song ◽  
Honglei Zhang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Primary Hyperparathyroidism ◽  
Secondary Hyperparathyroidism ◽  
Real Time ◽  
Real Time Pcr ◽  
Theoretical Basis ◽  
Down Regulation ◽  
Expression Characteristics

Abstract OBJECTIVE: Calcium-sensitive receptor (CASR) play an important role in the pathogenesis and progression of secondary parathyroidism (SHPT). The purpose of this study is to study the protein and gene expression characteristics of CASR in SHPT.Methods: Immunohistochemistry and real-time PCR were used to detect the expression of CASR protein and genes in SHPT tissues, and compared with the expression of CASR in primary hyperparathyroidism (PHPT).Results: CASR protein was down-regulated in SHPT and PHPT compared with normal parathyroid tissure（2.42±0.5 vs 3.2±0.62, P <0.05; 1.8±0.83 vs 3.2±0.62, P <0.05）, meanwhile SHPT tissue was higher than PHPT in the expression of CASR protein（2.42±0.5 vs 1.8±0.83, P <0.05）; CASR expression in SHPT tissue was higher than PHPT in gene expression（0.29±0.23 vs 0.01±0.12, P <0.05）. Conclusion: The expression of CASR protein and gene in SHPT is higher than that of PHPT. This feature provides a theoretical basis and ideas for studying the mechanism of CASR down-regulation.

scholarly journals The Effect of Interval Training on the Expression of Tumor Suppressor Gene, Systemic Inflammation, and Tumor Volume in Breast Cancer–Bearing BALB/c Mice

2019 ◽  
Vol 12 (3) ◽  
pp. 17-25
Author(s):  
Faranak Sadeghipoor Vojdani ◽  
Hamid Agha-Alinejad ◽  
Mahdiyeh Molanouri Shamsi ◽  
Sara Soudi ◽  
Sana Khanchi
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Tumor Suppressor ◽  
Tumor Suppressor Gene ◽  
Systemic Inflammation ◽  
Interval Training ◽  
Suppressor Gene ◽  
Exercise Session ◽  
Significant Difference ◽  
E Cadherin

Introduction: E-cadherin is expressed in most normal epithelial tissues. Loss of E-cadherin can cause dedifferentiation and invasiveness in human carcinomas, leading E-cadherin to be classified as a tumor suppressor. Therefore, the aim of this study was to investigate the effect of interval training on the expression of tumor suppressor gene E-cadherin in breast cancer-bearing BALB/c mice. Methods: Twenty female BALB/c mice were purchased from the Pasteur Institute and transferred to the Animal Laboratory of Tarbiat Modares University and randomly divided into four groups of training-cancer-rest, training-cancer-training, rest-cancer-rest, and rest-cancer-training. Cancer was induced by subcutaneous injection of 4T1 cell line. The mice performed an average-intensity interval training for 10 weeks, 5 days per week. Forty-eight hours after the last exercise session, the mice were sacrificed to measure the research variables. Gene expression was investigated using real-time PCR. The Kruskal-Wallis test was used to determine the statistical differences between groups. Results: There was a significant difference in the E-cadherin gene expression between the training-cancer-training and rest-cancer-rest groups (p = 0.03). This difference was also observed between the training-cancer-training and training-cancer-rest groups (p = 0.04). Conclusion: Interval exercise training may influence the expression of major tumor suppressor genes and systemic inflammation involved in the development of metastasis and even reverse this process.

scholarly journals Intestinal dysmotility after bowel resection in rats is associated with decreased ghrelin and vimentin expression and loss of intestinal cells of Cajal

2020 ◽  
Author(s):  
Igor Sukhotnik ◽  
Yoav Ben-Shahar ◽  
Yulia Pollak ◽  
Shlomi Cohen ◽  
Hadar Moran-Lev ◽  
...  
Keyword(s):  
Gene Expression ◽  
Protein Expression ◽  
Real Time ◽  
Real Time Pcr ◽  
Intestinal Motility ◽  
Bowel Resection ◽  
Down Regulation ◽  
Intestinal Transit Time ◽  
Protein Levels ◽  
Cells Of Cajal

The purpose of this study was to evaluate the mechanisms of intestinal motility in a rat model of short bowel syndrome (SBS). Rats were divided into three groups: Sham rats underwent bowel transection; SBS-NSI rats underwent a 75% bowel resection and presented with normal intestinal size (NSI) at sacrifice and hypermotility patterns; SBS-DYS showed dysmotile (DYS) enlarged intestine and inhibited motility patterns. Animals were sacrificed after 2 weeks. Illumina's Digital Gene Expression (DGE) analysis was used to determine the intestinal motility-related gene expression profiling in mucosal samples. Intestinal motility-related and interstitial cells of Cajal (ICC) genes and protein expression in intestinal muscle layer were determined using Real Time PCR, Western blotting and immunohistochemistry. Gastrointestinal tract motility was studied by microcomputer tomography. From ten Ca2+ signaling pathway related genes, six genes in jejunum and seven genes in ileum were down-regulated in SBS vs Sham animals. Down regulation of TMEM16A mRNA and protein was confirmed by Real Time PCR. Rapid intestinal transit time in SBS-NSI rats correlated with mild decrease in TMEM 16A, c-kit and vimentin mRNA and protein expression (vs Sham animals). SBS-DYS rats demonstrated enlarged intestinal loops and delayed small intestinal emptying (on imaging studies) that were correlated with marked down-regulation in TMEM 16A, c-kit, vimentin, ghrelin mRNA and protein levels compared to the other two groups. In conclusion, two weeks following massive bowel resection in rats, impaired intestinal motility was associated with decreased vimentin and ghrelin gene and protein levels as well as loss of ICC (c-kit and TMEM16A).

Ataxia Telangiectasia Mutated (ATM) Gene Expression Is Reversibly Up-Regulated in FANCA-Mutant Patient Fibroblasts by an Indirect Mechanism.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 1679-1679
Author(s):  
Johnson M. Liu ◽  
Abdallah Nihrane ◽  
Kazuhiko Yamamoto
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Real Time Pcr ◽  
Bone Marrow Failure ◽  
Wild Type ◽  
Down Regulation ◽  
Hematopoietic Stem ◽  
Primary Fibroblasts ◽  
Atm Gene ◽  
Atm Protein

Abstract Fanconi anemia (FA) predisposes to hematopoietic failure, birth defects and cancer, particularly leukemia and squamous cell carcinomas involving the cervix or head and neck. Although the FANC proteins have been shown to be involved in DNA repair, the pathogenesis of bone marrow failure and other developmental abnormalities is still unclear, but presumably, reflects abnormalities in senescence and apoptosis. The canonical ATM-p53 pathway is a pivotal mediator of the DNA damage response, which may act as a barrier to cancer through cell cycle and apoptosis regulation but has not been fully studied in FA. To address this, we first analyzed the integrity of the ATM-p53 axis in primary fibroblasts derived from FA patients. By immunoblot assays, we found that expression and phosphorylation of ATM and p53 were all up-regulated in two different FANCA-mutant fibroblasts following irradiation (IR) or mitomycin C (MMC) treatment, when compared to identically treated isogenic mutant cells transduced with wild-type FANCA cDNA. We reasoned that these results might be explained by up-regulation of the basal levels of ATM protein in the mutant fibroblasts. By real-time PCR, we confirmed that ATM gene expression was increased in the mutant line in comparison to the gene-corrected control. We also confirmed the up-regulation of both ATM protein and transcription in a FANCA-mutant EBV-immortalized lymphoblastoid cell line in comparison to its gene-corrected control. In order to determine if these changes in ATM gene expression were directly caused by FANCA depletion, we transfected HCT116 cells (known to be p53 wild-type) with two different siRNAs against FANCA. For these experiments, we used a plasmid expression vector (OriGene Technologies, Rockville MD) that drives 29-mer short hairpin RNAs targeting FANCA. In both transient and stable transfection experiments with either siRNA, we found that knockdown of FANCA expression was associated with down-regulation of ATM expression, as assessed by real-time PCR, when compared to cells transfected with an empty vector control. These results have led us to hypothesize that knockdown of FANCA leads to down-regulation of ATM, whereas cells with mutations in FANCA up-regulate ATM, perhaps as a result of chronic genotoxic stress not seen in the knockdown cells. Up-regulation of the ATM-p53 axis, in turn, may contribute to the hypo-proliferative characteristics seen in both primary fibroblasts and hematopoietic stem cells from FA patients.

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