Relationship study of the verified human epidermal growth factor receptor 2 amplification with other tumor markers and clinicohistopathological characteristics in patients with invasive breast cancer, using chromogenic in situ hybridization

Objective: Human epidermal growth factor receptor 2 (HER-2), as a crucial factor involved in about 20 of breast cancer cases, is one of the most reliable tumor markers to determine prognosis and therapeutic trend of this disease. This marker is generally assessed by immunohistochemistry (IHC) technique. In the cases that result of IHC test cast doubt (+2), the test should be repeated or validated by applying in situ hybridization techniques, like chromogenic in situ hybridization (CISH). In this regard, the goal of current study was to figure out the link between different clinicopathological characteristics of patients suffering from invasive breast cancer, using tumor markers, hormone receptor (HR) and HER-2. Comparing IHC and CISH techniques for evaluating diagnostic value and usefulness of HER-2 were also the other objective of this study. Materials and Methods: Based on this retrospective study, histological markers of 113 individuals suffering from invasive breast cancer -such as estrogen receptor (ER), progesterone receptor, HER-2 receptor, E-cadherin, CK5/6, vimentin and Ki67 were examined by IHC technique. HER-2 amplification of all patients was also evaluated by CISH. Clinicopathological information of the patients was also extracted from medical documents and their associations with tumor markers were statistically evaluated. Results: There is a significant relationship between tumor size, CK5/6 and tumor grade with HR status. Similar relationship was observed between HER-2 status and HR status, as well as vascular invasion (P<0.05). The comparison of HER-2 amplification showed no complete concordance of the result obtained from these two techniques, with score +3. Conclusion: Since the status of HER-2 is very important in decision making of the treatment process, CISH technique is recommended in the malignant conditions as the primary test, instead of IHC. In this study, we also determined that HER-2 expression is greatly correlated with ER- and PR- status. This might propose a better prognosis for HER-2+ patients. © 2019 Royan Institute (ACECR). All rights reserved

Chromosomal radiosensitivity and acute radiation side effects after radiotherapy in tumour patients - a follow-up study

Radiotherapists are highly interested in optimizing doses especially for patients who tend to suffer from side effects of radiotherapy (RT). It seems to be helpful to identify radiosensitive individuals before RT. Thus we examined aberrations in FISH painted chromosomes in in vitro irradiated blood samples of a group of patients suffering from breast cancer. In parallel, a follow-up of side effects in these patients was registered and compared to detected chromosome aberrations. METHODS: Blood samples (taken before radiotherapy) were irradiated in vitro with 3 Gy X-rays and analysed by FISH-painting to obtain aberration frequencies of first cycle metaphases for each patient. Aberration frequencies were analysed statistically to identify individuals with an elevated or reduced radiation response. Clinical data of patients have been recorded in parallel to gain knowledge on acute side effects of radiotherapy. RESULTS: Eight patients with a significantly elevated or reduced aberration yield were identified by use of a t-test criterion. A comparison with clinical side effects revealed that among patients with elevated aberration yields one exhibited a higher degree of acute toxicity and two patients a premature onset of skin reaction already after a cumulative dose of only 10 Gy. A significant relationship existed between translocations in vitro and the time dependent occurrence of side effects of the skin during the therapy period. CONCLUSIONS: The results suggest that translocations can be used as a test to identify individuals with a potentially elevated radiosensitivity

Chromosomal radiosensitivity in head and neck cancer patients: evidence for genetic predisposition?

The association between chromosomal radiosensitivity and genetic predisposition to head and neck cancer was investigated in this study. In all, 101 head and neck cancer patients and 75 healthy control individuals were included in the study. The G2 assay was used to measure chromosomal radiosensitivity. The results demonstrated that head and neck cancer patients had a statistically higher number of radiation-induced chromatid breaks than controls, with mean values of 1.23 and 1.10 breaks per cell, respectively (P<0.001). Using the 90th percentile of the G2 scores of the healthy individuals as a cutoff value for chromosomal radiosensitivity, 26% of the cancer patients were radiosensitive compared with 9% of the healthy controls (P=0.008). The mean number of radiation-induced chromatid breaks and the proportion of radiosensitive individuals were highest for oral cavity cancer patients (1.26 breaks per cell, 38%) and pharynx cancer patients (1.27 breaks per cell, 35%). The difference between patients and controls was most pronounced in the lower age group (⩽50 years, 1.32 breaks per cell, 38%) and in the non- and light smoking patient group (⩽10 pack-years, 1.28 breaks per cell, 46%). In conclusion, enhanced chromosomal radiosensitivity is a marker of genetic predisposition to head and neck cancer, and the genetic contribution is highest for oral cavity and pharynx cancer patients and for early onset and non- and light smoking patients

Study of the effect of MMC on the sister chromatid exchange in the human lymphocytes

Some environmental mutagenic agents cause genomic instability and increase susceptibility of DNA damage. One of them is mitomycin C which is connected to DNA as an alkylating factor and affects susceptible cells to reduction reactions. This drug is used in chemotherapy and treatment of tumors. Study of genomic instability in the presence of different concentrations of MMC can show susceptibility of DNA damage in the patients who are under chemotherapy with this drug. For this purpose, SCE is a qualified method that shows the number of sister chromatid exchanges in the metaphasic chromosomes. The number of 10^5 lymphocytic cells which were separated with ficol, were cultured in media (5ml, F12 15%-20% FCS) that contains mitogen of PHA (Phytohemagglutinin) and MMC in the concentrations of 3 ng/ml, 6 ng/ml and 9 ng/ml and a control sample without MMC. The specific concentration of BrdU was added after 24 hours to cell cultures. Then metaphasic cells were halted in the metaphasic stage with colchicine after 48 hours and were stained with SCE method and were studied for the number of sister chromatid exchanges in each metaphasic plaques. Evaluation of 100 metaphasic plates showed that SCE was 3.35% in the control cells while it was 5.43%, 7.1% and 8.13% in the treated cells with MMC in the concentrations of 3 ng/ml, 6 ng/ml and 9 ng/ml. In view of the results, it is clear than MMC can causes genomic instability even in the low concentrations and it can increase SCE so that the level of SCE is become the most with the concentration of 9 ng/ml and the least with the concentration of 3 ng/ml. In view of relation between SCE and DNA damage, we can conclude that the genome of normal cells will be damaged in the presence of MMC and in the patients who are under chemotherapy with this drug. It means that the genome of cells will become sensitive to mutation in the presence of low concentrations of MMC. Therefore we can postulate that we should use the concentrations of less than 3 ng/ml in order to decrease mutagenic effects of MMC in normal cells. Keywords: MMC, SCE, Cancer, Sister chromatid exchang

Evaluation of Radiation Sensitivity in Patients with Hyper IgM Syndrome

Background: HIGM syndrome is a rare form of primary immunodeficiencies characterized by normal/increased amounts of serum IgM and decreased serum levels of other switched immunoglobulin classes. Since the affected patients are continuously infected with various types of pathogens and are susceptible for cancers, diagnostic and therapeutic tests including imaging techniques are recommended for the diagnosis and treatment of these patients, which predispose them to higher accumulated doses of radiation. Given the evidence of class switching recombination machinery defect and its association with an increased rate of DNA repair, we aimed to evaluate radiation sensitivity among a group of patients diagnosed with HIGM syndrome. Methods: 19 HIGM patients (14 CD40 L and 3 AID deficiencies and 2 unsolved cases without known genetic defects) and 17 control subjects (10 healthy subjects as negative control group, 7 ataxia-telangiectasia patients as positive control group) were enrolled. G2 assay was carried out for the determination of radiosensitivity. Results: Based on radiation-induced chromosomal changes among the studied HIGM patients and their comparison with the controls, almost all (95) the patients had degrees of radiosensitivity: 6 patients with low to moderate, 1 patient with moderate, 11 patients with severe and 1 patient without radiation sensitivity. Conclusion: Today, X-ray radiation plays a very important role in diagnostic and therapeutic procedures; while increased exposure has devastating effects especially in radiosensitive patients. Considering higher sensitivity in HIGM patients, utilizing radiation-free techniques could partly avoid unnecessary and high-level exposure to radiation, thus preventing or reducing its harmful effects on the affected patients. © 2020, © 2020 Taylor & Francis Group, LLC

Proteomic analysis in human fibroblasts by continuous exposure to extremely low-frequency electromagnetic fields

Most people are Exposed to Extremely Low-Frequency Electromagnetic Fields (ELF-EMF). A number of studies have indicated association between exposure to extremely low frequency electromagnetic fields and a variety of cancers. Recently some therapeutic techniques such as repetitive Transcranial Magnetic Stimulation (rTMS) have been used to study localization of brain function, connectively of brain regions and pathophysiology of neuropsychiatric disorders (rTMS utilize low frequency-electromagnetic field). Here, the effect of continuous ELF electromagnetic fields (3 Hz, sinusoidal, 3 h and 4 mT) on the protein expression of human fibroblast cells is investigated via proteomics. The comparison of the 2-DE separated proteins from the exposed and sham (control) cells showed that some protein expressions are affected by radiation. The two proteins that their expression are reduced about 50 are determined as alpha 1 antitrypsin (A1AT) and Transthyretin (TTR). As it is reported that the amounts of these proteins reduced in the pathological conditions it can be concluded that application of ELF-EMF in therapeutic aspects may be to accompanying with their side effects. © 2007 Asian Network for Scientific Information