Genes copy number as a marker of low-invasive assessment of rectal tumors radiotherapy effectiveness.

2021 ◽  
Vol 39 (15_suppl) ◽  
pp. 3025-3025
Author(s):  
Marina A. Gusareva ◽  
Natalia G Kosheleva ◽  
Natalya B. Fatkina ◽  
Anna A. Solntseva ◽  
Lyudmila Ya. Rozenko ◽  
...  
Keyword(s):  
Blood Plasma ◽  
Copy Number ◽  
Tumor Regression ◽  
Extracellular Dna ◽  
Molecular Characteristics ◽  
Rectal Tumors ◽  
Number Variation ◽  
Group 2 ◽  
Low Efficiency ◽  
Group 1

3025 Background: Radiotherapy (RT) is a key component of rectal cancer (RC) treatment, however, nonresponsiveness in patients to preoperative RT is very common, usually due to the tumor cells radioresistance, mediated by their molecular characteristics, such as gene expression. The features of mRNA rapid degradation in extracellular environment make this indicator unsuitable for low invasive diagnostics. The solution to this problem is possible by switching to a more stable marker - the copy number variation (CNV), which can be determined in the extracellular DNA (cfDNA) circulating in the blood plasma. Therefore, the aim of the study was to identify the relationship between the level of genes CNV in the cfDNA of blood plasma with the effectiveness of rectal tumors RT. Methods: We used cfDNA preparations from blood plasma obtained before RT from 200 patients with RC, as well as from blood plasma of 50 apparently healthy donors (AHD, without cancer). RT was carried out on a linear accelerator Novalis TX (SFD = 2.4 Gy, TFD = 54.0 Gy). Blood samples were separated into plasma and cell fraction by centrifugation. Isolation of cfDNA from blood plasma was performed using the phenol-chloroform method. Determination CNV of 5 genes (BRCA2, H2AX, CASP9, RBBP8 and BCL2) was performed using Real-Time qPCR method. Differences were assessed using Mann-Whitney test; the Bonferroni correction was used to correct multiple comparisons. Results: RT results for 200 patients allowed them to be divided into 2 groups. After RT, 120 patients showed complete tumor regression (group 1), 50 patients showed insignificant tumor regression and 30 patients did not regress (group 2). In cfDNA of group 1 patients was found CNV decrease (p < 0.05) of H2AX and RBBP8 genes by 2.5 and 2.0 times, respectively, relative to AHD group. In the cfDNA of group 2patients an increase (p < 0.05) of BRCA2, H2AX, RBBP8 and BCL2 genes CNV was found by 2.0, 2.2, 2.0 and 2.0 times, respectively, relative to AHD group. Only 2 genes CNV differed in group 1 from group 2: the CNV of H2AX and RBBP8 was 5.4 and 4.0 times less respectively (p < 0.005). Conclusions: Thus, it has been found that increased CNV of genes BRCA2, H2AX, BCL2, RBBP8 in blood plasma cfDNA is associated with low efficiency of RT. At the same time, the CNV of H2AX and RBBP8 genes in cfDNA of patients with RC has the greatest potential as a marker of the RT effectiveness.

Association of genes copy number in extracellular DNA in patients with prostate cancer and sensitivity to radiotherapy.

2021 ◽  
Vol 39 (15_suppl) ◽  
pp. e15014-e15014
Author(s):  
Denis S. Kutilin ◽  
Mikhail S. Zinkovich ◽  
Marina A. Gusareva ◽  
Aleksandr V. Faenson ◽  
Elena A. Karnauhova ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Blood Plasma ◽  
Copy Number ◽  
Extracellular Dna ◽  
Biochemical Relapse ◽  
Prostate Tumors ◽  
Molecular Features ◽  
Number Variation ◽  
Invasive Method ◽  
High Level

e15014 Background: Radiotherapy (RT) is one of the main treatments for prostate cancer (PC). The effectiveness of such therapy depends on the initial radioresistance of tumor cells, which is ensured by their certain molecular features, which include the genes copy number variation (CNV). Model experiments on cell cultures (obtained from surgical material) have shown that CNVs have high potential as predictors of RT sensitivity. However, this potential is limited by the high level of invasiveness in obtaining biomaterials. A possible solution to this problem lies in the transition to CNV study in the extracellular DNA (cfDNA) of blood plasma. The aim of the study was to screen predictors of radioresistant PC based on the genes CNV in cfDNA. Methods: The study included 400 patients with diagnosed PC (T2a-3bN0M0, st. II-III), 40 of them after RT had a state of biochemical relapse (RT was performed on a Novalis TX linear accelerator (Varian, USA) (TFDisoeff = 75 Gr), mean time to biochemical relapse 7.5 months). Blood samples were separated into plasma and cell fraction by centrifugation. Isolation of cfDNA from blood plasma was performed using a set of reagents “DNA-Plasma-M” (Russia). Determination of the relative CNV of 13 genes (CDK1, CCND3, CDKN1B, TP53, PTEN, BCL2, XRCC4, BAX, RBBP8, H2AX, BRCA2, RAD50, EP300) was performed using the Real-Time qPCR method. Differences were assessed using the Mann-Whitney test; the Benjamin-Hochberg correction was used to correct multiple comparisons. Results: In the group with biochemical relapse (n = 40), the CNV of genes CDK1, CDKN1B, RBBP8, XRCC4, BRCA2 and RAD50 was statistically significantly (p < 0.05) higher by 2.0 times, 2.3 times, 2.1 times, 1.4 times, 2.4 times and 2.8 times, respectively, relative to the CNV of these genes in the cfDNA of the group without relapse (n = 360). Conclusions: Thus, it was found that the CNV of 6 genes (CDK1, CDKN1B, RBBP8, XRCC4, BRCA2 and RAD50) may be a potential molecular marker of radiosensitivity of prostate tumors. Based on the obtained data, a low invasive method for determining the prostate tumors sensitivity to RT has been developed.

scholarly journals Carcinosarcoma of the prostate: case report with molecular and histological characterization

2018 ◽  
Vol 33 (4) ◽  
pp. 540-544 ◽  
Author(s):  
Samanta Salvi ◽  
Valentina Casadio ◽  
Filippo Martignano ◽  
Giorgia Gurioli ◽  
Maria Maddalena Tumedei ◽  
...  
Keyword(s):  
Copy Number Variation ◽  
Copy Number ◽  
Copy Number Variations ◽  
Molecular Characteristics ◽  
Glutathione S Transferase ◽  
Molecular Alterations ◽  
Positive Expression ◽  
Molecular Pattern ◽  
Programmed Death ◽  
Number Variation

Background: We report a case of prostatic carcinosarcoma, a rare variant of prostatic cancer, which is composed of a mixture of epithelial and mesenchymal components with a generally poor outcome. Aims and methods: We aim to identify molecular alterations, in particular copy number variations of AR and c -MYC genes, methylation and expression of glutathione S-transferase P1 (GSTP1), programmed death-ligand 1 (PD-L1), AR, and phosphorylated AR expression. Results: We found a distinct molecular pattern between adenocarcinoma and carcinosarcoma, which was characterized by high AR copy number variation gain; positive expression of PD-L1, AR, and phosphorylated AR; low espression of GSTP1 in epithelial component. The sarcomatoid component had a lower gain of the AR gene, and no expression of PD-L1, AR, phosphorylated AR, or GSTP1. Both components had a gain of c-MYC copy number variation. Conclusions: Our findings suggest that carcinosarcoma has specific molecular characteristics that could be indicative for early diagnosis and treatment selection.

Clove (Syzygium aromaticum) Effect on Carbon Tetrachloride-Induced Rat: Comparison of Malondialdehyde Level of Liver and Blood Plasma

2018 ◽  
Vol 24 (8) ◽  
pp. 6053-6057
Author(s):  
Muhammad K Azwar ◽  
Ani R Prijanti
Keyword(s):  
Carbon Tetrachloride ◽  
Blood Plasma ◽  
Antioxidant Properties ◽  
Oxidative Stress Marker ◽  
Control Group ◽  
Syzygium Aromaticum ◽  
Group 4 ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

Studies suggested antioxidant properties of the content of Syzygium aromaticum (clove). The study was conducted to obtain better understanding about the effect of clove on concentration of oxidative stress marker malondialdehyde (MDA) in liver and blood plasma of rat initially induced by carbon tetrachloride (CCl4); and whether blood plasma MDA level might represent liver condition. Experimental research was done using 20 Wistar rats classified into 5 treatment groups: (1) CCl4 - and clove-positive treatment after 3 days of clove treatment, (2) one day after, (3) alpha-tocopherol as positive control, (4) CCl4 only as negative control, and (5) normal control. Wills method was used for MDA concentration measurement. Liver MDA concentration were 0.0262 ± 0.0010 for group 1, 0.0214 ± 0.0047 group 2, 0 for group 3, 0.0077 ± 0.0094 group 4, and 0.0039 ± 0.0009 control group in nmol/mg protein (p = 0.001), whereas in the blood plasma it was 29.6032 ± 6.8021 for group 1, 26.1103 ± 3.6920 for group 2, 1.1612 ± 0.3555 for group 3, 1.4585 ± 1.4747 for group 4, and 2.4217 ± 1.2382 control group in nmol/mL (p = 0.001). Contrary to study in the past, no antioxidant properties were observed in treatment with dose 200 mg clove/kg body weight of rat. Such treatment increased MDA concentration and enhanced CCl4-induced damage in a time-dependent fashion. Strong correlation between MDA concentration in the liver and blood plasma (R = 0.97; p = 0.003) suggested blood plasma utilisation to represent hepatic MDA concentration or damage.

scholarly journals Blood Plasma Serotonin and von Willebrand Factor as Biomarkers of Unstable Angina Progression Toward Myocardial Infarction

2021 ◽  
Vol 28 (1) ◽  
pp. E202112
Author(s):  
Yuliya Tyravska ◽  
Oleksandr Savchenko ◽  
Viktor Lizogub ◽  
Nataliia Raksha ◽  
Olexiy Savchuk
Keyword(s):  
Myocardial Infarction ◽  
Blood Plasma ◽  
Von Willebrand Factor ◽  
Serotonin Concentration ◽  
Group 3 ◽  
Group 2 ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
Plasma Serotonin ◽  
Group 1

Aim: To investigate the serotonin and von Willebrand factor (vWF) concentrations among unstable angina (UA) patients without and with progression toward myocardial infarction (outcome) and to assess the utility of both as prognostic markers of UA complications. Materials and methods: In observational cohort study, we recruited 103 patients with ischemic heart disease (the median age 65.0 (59.0-69.0) years, 45 females (43.7%)). After full set of investigations including high sensitive Troponin I test and 28-day follow-up period, we defined three groups: Group 1 - stable angina patients (n=22) as control, Group 2 - UA patients without outcome (n=71), Group 3 - UA patients with outcome (n=10). We analyzed the blood plasma serotonin content by the ion-exchange chromatography with measurement of serotonin on fluorescence spectrophotometer. VWF concentration was determined by ELISA. We compared the concentrations of observed parameters among the groups with the Kruskal-Wallis test (with post-hoc Mann-Whitney test with Bonferroni-Holm correction). We assessed binary logistic models, receiver operating characteristic curves, calculated sensitivity (Se), specificity (Sp), and positive likelihood ratio (LR+) for each indicator. Results: We registered elevation in serotonin concentration and decline in vWF concentration in Group 3 in comparison with Group 2 (22.670 (20.687-24.927) μg/ml vs 11.980 (8.120-15.000) μg/ml, p< 0.001, and 0.117 (0.109-0.120) rel.units/ml vs 0.134 (0.127-0.143) rel.units/ml, p < 0.001) and Group 1 (12.340 (10.052-13.619) μg/ml, p < 0.001, and 0.137 (0.127-0.156) rel.units/ml, p < 0.001), respectively. No significant differences in serotonin and vWF concentrations between Group 1 and Group 2 were detected (p=0.81 and p=0.36, respectively). The probability of outcome increased significantly (by 60.7% and 59.7%, LR+ 19.0 [6.0, 60.0] and 18.0 [3.9, 80.0]) if serotonin concentration was above 21.575 μg/ml (Se=80.0%, Sp=95.8%, AUC=0.975) and vWF concentration was below 0.114 rel.units/ml (Se=50.0%, Sp=97.2%, AUC=0.973), respectively. Conclusions: Serotonin and vWF as biomarkers are demonstrated promising results for rule-in the patients with risk of short-term UA progression toward myocardial infarction.

Cobimetinib plus gemcitabine is an active combination in KRAS G12R-mutated in previously chemotherapy-treated and failed pancreatic patients.

2020 ◽  
Vol 38 (15_suppl) ◽  
pp. 4642-4642
Author(s):  
Bach Ardalan ◽  
Jared Addison Cotta ◽  
Miriam Gombosh ◽  
Jose Ignacio Azqueta
Keyword(s):  
Pancreatic Cancer ◽  
Cancer Patients ◽  
Tumor Regression ◽  
Mapk Pathway ◽  
Progression Free Survival ◽  
Mek Inhibitor ◽  
Kras Mutations ◽  
Positive Study ◽  
Group 2 ◽  
Group 1

4642 Background: he KRAS proto-oncogene is involved in the RAS/MAPK pathway. Various G12X mutations have been examined with the most common mutations being G12D (40%), G12V (30%), and G12R (15-20%) in pancreatic cancer patients. Throughout the course of studying the G12X mutations, we have observed that not all KRAS mutations are equal. Preclinical data shows G12R is impaired in pI3Kα signaling, as compared to KRAS G12V/D. This mechanism is important in PDAC as it allows tumor growth to be sustained. In preclinical studies, PDX derived tumors were transplanted in mice and were treated with a MEK inhibitor plus chemotherapy, which demonstrated a greater tumor regression than either agent alone. Therefore, we have decided to treat patients with Gemcitabine alongside a 2nd generation MEK inhibitor (Cobimetinib). Methods: In our single arm study, 13 KRAS mutated pancreatic patients (KRAS G12D, G12V, and G12R) received the combination of Cobimetinib 20mg BID weekly for three weeks alongside Gemcitabine at 1000mg/m2 weekly, followed by one week of rest. The above constitutes one cycle. Results: Patients were divided into two groups; Group 1 consists of seven patients that were KRAS G12D/G12V mutated, and Group 2 included six KRAS G12R mutated patients. In Group 1, seven patients on treatment progressed and died within two months on the study. In Group 2, one achieved PR and others stable disease. Median progression-free survival was 6.0 months (95% CI 3-9.3 months) and median OS has not been reached. All patients are alive at 8 months. Common adverse reactions include rash, fatigue, nausea, and vomiting. Cancer antigen 19-9 decreased in ≥ 50 of all patients in the latter group. We would like to report our positive study to the society. Moreover, we intend to confirm the study in a larger patient cohort. Conclusions: Pancreatic cancer patients that demonstrate KRAS G12R mutations are treatable with a new active combination chemotherapy.

scholarly journals 161 PREGNANCY RATES OBTAINED AFTER EMBRYO TRANSFER AT FIXED TIME OF IN VIVO-, IVF- AND CLONED-DERIVED EMBRYOS

2005 ◽  
Vol 17 (2) ◽  
pp. 231
Author(s):  
J. Lagioia ◽  
M. Panarace ◽  
M. Marfil ◽  
M. Basualdo ◽  
J. Gutierrez ◽  
...  
Keyword(s):  
Pregnancy Rate ◽  
Embryo Transfer ◽  
Fixed Time ◽  
Pregnancy Rates ◽  
Transfer Programs ◽  
Estrus Detection ◽  
Group 2 ◽  
Low Efficiency ◽  
Group 1

The most important factor in bovine embryo transfer programs is the low efficiency in the utilization of the recipients; this low efficiency is associated with low response to synchronization protocols and failures in estrus detection. It has been shown that cows transferred at fixed time with in vivo-derived embryos resulted in high rates of recipients selected for transfer and high overall pregnancy rates (recipients pregnant/recipients treated) (Tribulo et al. 2002 Theriogenology 57, 563). An experiment was designed to evaluate the pregnancy rate in recipients transferred with in vivo (fresh and frozen), IVF, and cloned-derived embryos without estrus detection. A total of 1555 non-lactating Bos Taurus crossbred beef cows was divided into two groups. Cows from group 1 (n = 421) were synchronized with a progesterone intravaginal releasing device (1 g P4; DIB, Syntex®, Buenos Aires, Argentina) plus 2 mg of estradiol benzoate (EB) i.m. (Syntex®) on Day 0. On Day 5, they received 400 IU of eCG (Novormon 5000, Syntex®) i.m. and 150 μg of D-Cloprostenol (PGF2α) (Bioprost-D, Biotay®, Buenos Aires, Argentina). The DIB devices were removed on Day 8 and on Day 9, 1 mg of EB was injected. Day 10 was arbitrarily considered as the day of estrus. Cows from group 2 (n = 1134) received 2 doses of PGF2α 14 days apart and were checked for heat during 5 days after the second PGF2α dose. Cows of both groups were examined 7 days after estrus by ultrasonography (Pie Medical Scanner 200®) and those with a corpus luteum >10 mm of diameter were transferred nonsurgically with in vivo (fresh and frozen), IVF, and cloned-derived embryos. In group 1, 360 cows were transferred, and in group 2, 726 cows were transferred (Table 1). Pregnancy was diagnosed 23 days later by ultrasonography (Pie Medical Scanner 200®). The pregnancy rates were compared statistically between groups 1 and 2 by analysis of variance (Infostat, LSD Fisher). There was no significant statistic difference (P > 0.05) between pregnancy rate in group 1 and 2 with in vivo (fresh), IVF, and cloned-derived embryos. However, pregnancy rate of frozen in vivo-derived embryos was lower in group 1 than in group 2 (P < 0.05). Results showed that treatment using DIB combined with EB, PGF2α, and eCG associated with embryo transfer without estrus detection (group 1) had no difference in pregnancy rate when compared with the treatment where synchronization with PGF2α and heat detection were used (group 2). Another important advantage is the use the group 1 treatment for increasing the flexibility and efficiency in the management of the recipients of in vivo, IVF, and cloned-derived embryo transfer programs. Table 1. Comparison of pregnancy rates between group 1 (embryo transfer at fixed time) and group 2 (embryo transfer 7 days after estrus detection)

Peculiarities of Fibrinolytic and Proteolytic Activity of Blood Plasma in Patients with Chronic Pancreatitis Combined with Hypothyroidism

2021 ◽  
Vol 6 (5) ◽  
pp. 233-238
Author(s):  
V. V. Ratsa ◽  
◽  
O. I. Fediv
Keyword(s):  
Molecular Weight ◽  
Chronic Pancreatitis ◽  
Blood Plasma ◽  
Fibrinolytic Activity ◽  
The State ◽  
Low Molecular Weight ◽  
Healthy Individuals ◽  
Group 2 ◽  
Low Molecular Weight Proteins ◽  
Group 1

The purpose of the study is to analyze the state of proteolytic and fibrinolytic activities of blood plasma in patients with chronic pancreatitis combined with hypothyroidism. Materials and methods. 105 people participated in our study, of which group 1 consisted of patients with chronic pancreatitis (n = 27), group 2 – patients with hypothyroidism (n = 30), group 3 – patients with chronic pancreatitis combined with hypothyroidism (n = 28), group 4 – almost healthy individuals (n = 20). The state of fibrinolytic activity of blood plasma was studied by lysis of azofibrin, followed by determination of total fibrinolytic activity, non-enzymatic fibrinolytic activity and enzymatic fibrinolytic activity. Assessment of the state of the proteolysis system was studied by lysis of azoalbumin (breakdown of low molecular weight proteins), azocasein (breakdown of high molecular weight proteins) and azocol (breakdown of collagen). Results. When analyzing the results of the study, we observe a probable increase in lysis of azoalbumin by 1.89, 1.96 and 2.16 times (p <0.05) in groups 1, 2, 3 compared with the group of almost healthy individuals. In patients with chronic pancreatitis and hypothyroidism, the most pronounced degradation of low molecular weight proteins was observed, which was 13.86% and 9.75% (p <0.05) higher than in the first and second groups. Indicators of azocasein lysis by 52.48%, 56.35% and 95.03% (p <0.05) were found in groups 1, 2, 3 compared with almost healthy individuals. Azocasein lysis was higher by 27.89% and 24.73% (p <0.05) in patients with chronic pancreatitis combined with hypothyroidism than in patients in groups 1 and 2. Azocol lysis was significantly higher by 10.85%, 12.05%, 16.87% (p <0.05) in groups 1, 2, 3 compared with almost healthy individuals. In addition, in patients with comorbid pathology there was an increase in lysis of azocol by 5.3% and 4.3% (p <0.05) compared with the first and second groups. The total fibrinolytic activity of blood plasma was 8.3%, 6.7%, 16.26% (p <0.05) lower in patients of groups 1, 2, 3 compared with almost healthy individuals. Non-enzymatic fibrinolytic activity of blood plasma was 44.89%, 49.64%, 66.27% higher in groups 1, 2 and 3 than in almost healthy individuals. Enzymatic fibrinolytic activity of blood plasma was 44.28%, 42.25%, 90.57% (p <0.05) lower in group 1, 2, 3 compared with the group of almost healthy individuals (p <0,05). There was a decrease in the level of enzymatic fibrinolytic activity of blood plasma by 32.07% and 33.96% (p <0.05) in patients with chronic pancreatitis associated with hypothyroidism compared with participants in groups 1 and 2 without comorbid pathology. Conclusion. The most pronounced changes in proteolytic (increased lysis of azoalbumin, azocasein, azocol) and fibrinolytic (decrease in total, non-enzymatic and enzymatic) activities of blood plasma in patients with chronic pancreatitis associated with hypothyroidism were determined

Intraoperative detection of lymph node metastasis using one-step nucleic acid amplification (OSNA) in breast cancer patients: Effect on second surgery rate and delay for adjuvant therapy.

2012 ◽  
Vol 30 (15_suppl) ◽  
pp. 10517-10517 ◽  
Author(s):  
Sophie Klingler ◽  
Frederic Marchal ◽  
Philippe Rauch ◽  
Ouarda Kenouchi ◽  
Anne-Sophie Chrétien ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Lymph Node ◽  
Adjuvant Therapy ◽  
Copy Number ◽  
Isolated Tumor Cells ◽  
Ck19 Mrna ◽  
Second Surgery ◽  
The Mean ◽  
Group 2 ◽  
Group 1

10517 Background: Sentinel lymph node (SLN) analysis is conventionally analysed using HES and CK19 immunohistochemistry. In case of SLN involvement, a second surgery is required for axillary lymph node (ALN) resection thus delaying the initiation of adjuvant therapies. Methods: 381 pts with invasive breast cancer were considered in this retrospective study. SLN were detected using combined radio-isotope and color detection. SLN involvement was analysed using OSNA for CK19 mRNA, in 100 pts (group 1) and compared to conventional histopathology in 281 pts (reference population, group 2). In all cases, control cytology was performed. Results: No significant difference was found between group 1 and 2 regarding patients characteristics, tumor localization, size, grade, steroid receptors and HER2 expression and the mean number of SLN analysed per pt, 2.4 (range 1-7) and 2.5 (range 1-8), respectively. Considering positive SLN as “++” (CK19 mRNA copy number>5000), “+” (250 < CK19 mRNA copy number < 5000) and positive by inhibition in group 1 and macro-, micrometastases and isolated tumor cells in group 2, no difference in lymph node involvement rate was found between the two groups with 29.0 and 29.9% of positive SLN, respectively. Only one discordance case was observed with negative OSNA analysis and positive cytology with isolated tumor cells. Using OSNA intraoperatively, the mean time to process SLN was 42 min (range 10-104) allowing immediate ALN resection. Significant (P<.01) reduction of re-intervention rate was observed when OSNA was used (9 vs 39%), including margins insufficiency (5 vs 13%), ALN resection (3 vs 15%) or both (1 vs 11%). Delay before adjuvant therapy was also significantly (P <.01) reduced when OSNA was used with 43 (range 20-82) vs 59 (range 14-212) days. Conclusions: Results achieved with OSNA are fully consistent with those achieved using conventional immunohistochemistry analysis. SLN analysis using OSNA avoids a second operation in most patients for ALN resection and shortens the delay for adjuvant therapy.

Copy number variation of MDM2 and p53 genes in extracellular DNA as potential marker for lung adenocarcinoma diagnostics.

2017 ◽  
Vol 35 (15_suppl) ◽  
pp. e23094-e23094
Author(s):  
Denis S. Kutilin ◽  
Yaroslav S. Enin ◽  
Dmitriy I. Vodolazhsky ◽  
Vyacheslav A. Sustretov ◽  
Tamara G. Ayrapetova ◽  
...  
Keyword(s):  
Lung Adenocarcinoma ◽  
Copy Number ◽  
Gene Copy Number ◽  
P53 Gene ◽  
Extracellular Dna ◽  
Gene Copy ◽  
Mdm2 Gene ◽  
Healthy Donors ◽  
Number Variation ◽  
P53 Genes

e23094 Background: Today, the problem of early diagnostics of lung cancer remains unsolved as every fourth patient diagnosed with the disease already has distant metastases. Studying gene copy number variation (CNV) in extracellular DNA in the blood plasma can become a basis for a new effective and low invasive method for predictive diagnostics and disease prognosis. The purpose of the study was to examine the copy number of the MDM2 and p53 genes in extracellular DNA of patients with metastatic and non-metastatic lung cancer and healthy donors. Methods: The blood samples of 30 patients with lung adenocarcinoma (collected before surgery) and 30 healthy donors (without cancer) were studied. Each sample was centrifuged to obtain blood plasma. DNA was isolated from plasma using a phenol-chloroform extraction method. Detection of relative copy number of the MDM2 and p53 genes (reference gene - GAPDH) was performed by RT-qPCR using CFX96 thermocycler (Bio-Rad, USA). The groups were compared by the Mann-Whitney U- test. Results: Reduction of the p53 gene copy number by 57% (p < 0.05), as well as increasing of the MDM2 gene copy number by 160% (p < 0.05), were found in the extracellular DNA of patients with lung adenocarcinoma compared with healthy donors. As a result, the ratio of copy number of pro-/anti-apoptotic genes p53:MDM2 in extracellular DNA of patients with lung adenocarcinoma (1:23 ratio) was different from that of healthy donors (1:4 ratio). The MDM2 gene copy number in extracellular DNA of patients with metastases exceeded the value in non-metastatic patients two-fold (p < 0.05). The p53 gene copy number in extracellular DNA of patients with metastatic and non-metastatic cancer did not differ significantly. Conclusions: CNV of the p53 and MDM2 genes in extracellular DNA has a high potential for low invasive diagnostics and prognosis of lung adenocarcinoma.

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