Iraqi Journal of Cancer and Medical Genetics

Alterations of tumor suppressor gene p53 were identified in a various types of human cancers, including bladder cancer. So, the present study was designed to investigate the relationship between the tumor suppressor gene p53 in some Iraqi patients with bladder cancer by using two methods, insitu hybridization compare with immunohistochemical assay. A series of 40 bladder tissue cancer collected from different sites (transitional cell and squamous cell) from patients who had undergone cystactomy. These samples were collected during the period between (2012-2014). The histologic types included 21 well differentiated transitional cell, squamous cell, 9 moderately differentiated and ten poorly differentiated carcinomas. By using in situ hybridization, tumor suppressor gene p53 was detected in eighteen out of fourty (45%), otherwise, at immunohistochemistry, it was detected in twenty eight out of fourty (70%). Moreover, the positive results of tumor suppressor gene p53 was related in highly significant to each of stages and sites , significantly related with histologic grade of the tumor by using insitu hybridization method, but there was no significant correlation with each of age and smoking patients. furthermore, in immunohistochemical assay p53 related in highly significant correlation with each of histologic grade, sites, stages of the tumor and the smoking habit. So, it was significantly related with age and gender. It could be conclouded from this study that the two methods of in situ hybridization and immunohistochemistry for detection of tumor suppressor gene , protein p53 is useful in the clinical evaluation of patients with bladder cancer and it suggested that mutations of p53 are involved in the bladder transitional cell and squamous cell carcinomas and it may play an a major role in the malignant transformation of urinary tract cancer.

This study was done to evaluation the cytotoxicity of each apricot kernels ethanolic extract at concentration (100-500) μg/ml, amygdalin and methotreaxat at concentration (0.1-1000) μg/ml, (plant extract plus methotreaxat) and (amygdalin plus methotreaxat) at half the original concentration for each one on Hela (human cervical cancer cell line) in vitro. A mixtures of (extract plus Methotreaxte) and (amygdalin plus Methotreaxte) was used together to evaluated its cytotoxicity on cancer cell line for indentify about the possibility of synergistic effect between them. The results show the cytotoxicity of (plant extract and methotreaxat) on Hela cancer cell line was same as the cytotoxicity of plant extract alone and more than the cytotoxicity of Methotreaxte alone, while the cytotoxicity of (amygdalin and methotreaxat) on Hela cancer cell line is similar to the cytotoxicity of amygdalin alone and more than the cytotoxicity of Methotreaxte alone.

Cancer targeted nanotherapy represent an exciting field in the search for new cancer specific therapies to avoid conventional chemotherapy side effects. Because cancer cells usually have malfunctioning apoptotic machinery which favors survival pathways and drug resistance. Cancer cell apoptosis is the favorable event to be induced in any new anticancer agent development. Nanotherapy goals are to elevate therapeutic efficiency, selectivity, and overcome drug resistance as major obstacle in cancer treatment. Hybrid nanoliposomes (nHLs) may fulfill all these features in cancer therapeutics. We have previously demonstrated the ability of in house synthesized nHLs to inhibit HeLa cell line proliferation and study preliminary the induction of apoptosis as a consequences of that inhibition. In order to confirm the event of apoptosis in HeLa cell line incubated with the synthesized nHLs we exposed HeLa cells to inhibition concentration 50 (IC50) of previously synthesized hybrid nanoliposomes. Mechanism of apoptosis induction was determined using mitochondrial membrane potential disruption, caspase-3 activity and single cell gel electrophoresis as well as DNA fragmentation assay. All apoptosis detection procedures used gave a clear defined significant indication that nHLs was capable of induce apoptosis in HeLa cells through intrinsic pathway. This result needs further investigation to confirm nHLs as potential nanotherapy.

Glutathion S-transferase enzymes are detoxifying enzymes – phase II enzymes, which play an important role in protecting cells from damage caused by endogenous and exogenous compounds and subsequently, are crucial in protecting the DNA. The aim of this study is to evaluate polymorphism of Glutathion S-transferase P1 (GSTP1) in Iraqi women breast cancer. Blood samples were collected from 65 Iraqi women diagnosed as breast cancer patients who attended to Oncology Teaching Hospital and Al-Yarmouk Teaching Hospital. In addition to 59 healthy women as control group. Genomic DNA was extracted then genotyping was performed by restriction fragment length polymorphism- polymerase chain reaction (RFLPPCR) for GSTP1 gene. The frequencies of GSTP1 polymorphism (Ile105Val) in the breast cancer group were: 41.54% for wild type (Ile/Ile), 49.23% for heterozygote (Ile/Val) and 9.23% for homozygote mutant (Val/Val), while in control group was 57.63 for wild (Ile/Ile), 42.37(Ile/Val) heterozygote and 0.00 %(Val/Val) homozygote. The results show there were statistically significant differences in the distribution of (Val/Val) genotype and Val allele between patients and controls groups (p<0.05), (OR= 16.3091; 95% CI= 0.8798 to 302.3326) and (OR= 1.9380; 95% CI= 1.0274 to 3.6556) respectively . This study showed that GSTP1 (Val/ Val) genotype may be associated with an increased risk for breast cancer among Iraqi women.

Many scientists have focused on the treatment of cancer for along time by studying cytotoxic activities of plant extracts , the E.spectabilis have been used in anticancer studies due to their phytochemical content included (carvone , carvacrol, pentane , 2-methyl , 9-caryophyllene). In present study, we demonstrated the cytotoxic activity of aqueous and Hexan _ ethanolic extracts on cell viability of Rabdumyosarcoma cell (RD) , and transformed kidney epithelium of African green monkey (Vero). In dose and time dependent manner Aqueous and hexan_ ethanol extracts (10.000,1000,100,10,1,0.1,0.01,0.001)μg/ ml at (24, 48, 72hrs.) were examined on RD and Vero cells. The result of current study showed that Aqueous and hexan_ethanol extracts of E. spectabilis effectively inhibited the cell proliferation by decreasing the cell viability of both cell line at different concentration, in which (RD) cell line was sensitive to all concentrations of the plant extract in this period, while the (Vero) cell line was resistant at the two concentrations which was (100 and 10)μg/ml with the means (0.080,0.16).While Hexan_ethanolic extract of different concentrations for (72hrs.) of exposure on both cell lines (Vero and RD) on cell growth assay show that the RD cell line was not respond to all the concentrations that used for killing of RD cell line while Vero cell line was represent its sensitivity for all concentrations from (10.000_0.001)μg/ml.

Background: In Iraq bladder carcinoma is a major problem according to the results of the Iraqi cancer registry between the years (1991-2006). Cancer of the urinary bladder was the 2nd on the table of the commonest ten cancers in male and it was the 8th commonest cancer in females (1). According to the same registry the main histological types were: Transitional cell carcinoma: 43.82%. Sequamous cell carcinoma: 4.49%. Undifferentiated cell carcinoma: 0.31 %. Adenocarcinoma: 0.92%. Male to female ratio was 3.39:1 the peak age incidence was found in the 70+ years (1). Bilharzial ovals were found in about 7%-29% of bladder cancers, which is lower than that in Egypt; 82-90% (2). On the other hand urinary tract infections are among the most common bacterial infections that lead patients to seek medical care. It had been estimated that more than 6 million outpatient visits and 300,000 hospital stays every year are due to urinary tract infections (3). The aim of this study is to find the association or disassociation between bacterial infection of the urothelium and urothelial carcinoma and if it was associated which type is it: causative or associative.
Material and Method: This study was carried out on 105 patients with carcinoma of the bladder (transitional, sequamous), whom were referred to the surgical specialties hospital, medical city for diagnostic transurethral endoscopy during the period between 5th of October 2007 till the 1st of September 2008. History of smoking, previous recurrent urinary tract infection and stone was taken, identification of the causative bacteria for their urinary tract infection was done from three samples collected; two urine samples (on admission and post-operatively) and one tissue sample (tumor biopsy).
Results: The results showed that: 85.65% of the patients had infections associated with their tumors, the rest 14.35% were negative. These infections were either Escherichia coli, Klebsiella, Proteus, Staphylococcus, Enterobacter, Citrobacter or Morganella spp. Escherichia coli was the most common 73.3% of all the cases. The results also showed that there was a significant association between positive urine culture on diagnosis for bacterial growth and urothelial carcinoma, also Escherichia coli was collected from the intracellular compartment by a mechanical approach.
Conclusion: Associations between urothelial carcinomas and bacterial growths were studied and showed that urothelial carcinomas were highly associated with bacterial growth especially Escherichia coli. Although Klebsiella, Proteus, Staphylococcus, Enterobacter, Citrobacter or Morganella spp., occured in a lower incidence, still had a role in urinary tract infection. Finding Escherichia coli intracellular and its high association with urothelial carcinoma could put these bacteria in the causative section of urothelial carcinoma through producing a recurrent acute infection.

Background: A number of reports have demonstrated the presence of HPV in endometrial adenocarcinomas(ECs) . alterations in Cyclin D1(CCND1) gene expression has been reported in ECs and the integration of human papilloma virus (HPV) could alter the level of gene expression of CCND1.
Methodology: This study has used in situ hybridization to localized HR-HPV16/18 in tissue specimens from 70 hysterectomized patients diagnosed with malignant endometrial tumors (30 cases), non-malignant endometrial tumors (25 cases), and 15 cases as control tissues groups. Also the study has enrolled detection the gene expression of CCND1 using Immunohistochemistry.
Results: In the cancer sites malignant endometrial tumors group, HR-HPV16/18 was detected in 10 cases(33.3%) ,and in 5 cases (20%) non-malignant endometrial tumors ,and 4 cases (26.7%)of control tissues group. Most HPV16/18 infections have Punctate DNA pattern .The results of moleculare detection of CCND1 revealed 13(43.3%) in malignant endometrial tumor 8(32%) non-malignant endometrial tumor and 6(40%) in control groups. The significant correlation between CCND1 expression and HR-HPV16/18 infection was reported only in malignant endometrial tumors groups.
Conclusions: HR-HPV16/18 may be associated with initiation of endometrial carcinogenesis events as well as play a role in the progression of such malignant tumors.CCND1 could have sharing in early events of tumorgenesis in endometrial carcinoma and a significant correlations of CCND1 expressions with HPV infection was observed among malignant endometrial tumor.

AN3 tumor model had special importance as breast cancer model to evaluate new and novel therapeutic agents and there is need to study the tumor associated markers presence during AN3 tumor growth. Most common markers are Carcinoembryonic antigen (CEA) and α-fetoprotein (AFP) which are cells surface glycoproteins. Oncofetal antigens involved in cell adhesion marker for cancers diagnosis and prognosis and normally they are not present in the blood of healthy adults. AN3 is transplantable mammary adenocarcinoma tumor used as in vivo model for years in cancer research for human breast cancer. Hence studying the presence of associated tumor antigens that extensively used as tumor markers (AFP and CEA) is necessary to characterize the model and determine the usefulness to use them as follow up markers after testing therapeutic agents in this model. Healthy and pregnant healthy female mice were used as control groups and AN3 transplanted mice measured after tumor grow to reach at least 0.5 cm. The results showed that CEA is markedly high in mice bearing tumors while no presence of it in both control groups. AFP had a different picture as the higher percentage was in the pregnant females, which is very normal because AFP is a fetal protein, but mice bearing tumor significantly higher concentration than healthy mice. In conclusion, AFP and CEA are useful markers to be used for AN3 model in experimental evaluation of novel therapies follow up.

Through the period April - December 2012, urine specimens were collected from186 patiants attending five hospitals. Seventy six isolates of uropathogenic E. coli were obtained from these samples. These isolates were identified as E. coli according to the cultural and morphological characteristics on MacConkey agar, eosin methylene blue agar, biochemical test (vitek 2 system), and API 20 E system. The results showed that only four E. coli isolates (E4, E8, E23, E36) produced cytotoxic necrosis factor 1(CNF1), and E23 was the more efficient isolate in CNF1 production and have high activity on tissue culture. CNF1 produced from this isolate was purified by two steps; the first was by ion exchange chromatography using DEAESephadex, and the second step was gel filtration by Sephacryl S-100. Also study Polymerase Chain Reaction (PCR) for cnf1 gene and used primer cnf1. The size of amplified gene was 543pb. The cytotoxic effect of different concentrations of CNF1 on cancer lines (RD, AMN3, , Hela, PC3 and HePG2) at 72 hr were examined .The effect was manifested multinucleated and enlargement in cell line. The low IC50 was 104μg/ml against HePG2cancer cells after 72 hr. Also, study multiparameter cytotoxic assay. The results revealed low concentration of cytochrome c, total nuclear intensity, cell membrane permeability in comparative positive control. The mitochondria concentration was high compared to the positive control on of cell lines in the concentration (100,25, 12.5,6.25).

Background: Acute lymphoblastic leukaemia accounts for approximately 14% of leukaemia cases in the adults with B linage subtype being more common than T linage subtype. immnophenotyping using the multicolour flow cytometry is an essential tool for diagnosis, classification as well as treatment guidance and prognosis.
Objectives: This study aims to provide an overview of the immunophenotypic profile of adult ALL cases in Iraq, it’s possible link with other characters and also study the frequency of expression of aberrant myeloid antigens in them.
Patients and methods: this is a descriptive study included 64 adult patients diagnosed as ALL in haematology unit at Baghdad teaching hospital for the period between May 2014 till May 2015 , immunophenotyping was carried out at the flow cytometry laboratory in the nursing home hospital using 6 coloured (BD-FACS-Canto II System from Becton Dickenson).
Results: of the study population 63.7% of the cases were B-ALL and 27.5% were T-ALL, there was no significant difference in respect to age and gender between them. Of the CD markers used both CD10 and CD34 were significantly associated with B-ALL compared to T-ALL (p values 0.002 and 0.003 respectively).aberrant myeloid Ag expression was seen in 43.18% of B-ALL cases and in 52.6% of T-ALL cases with no significant difference however aberrant myeloid Ag expression was significantly associated with CD34 expression with a p value of 00.018.
Conclusion: Immunophenotyping retains a crucial place in the work up for patients with acute lymphoblastic leukaemia. CD34 was significantly associated with B linage ALL and with aberrant myeloid Ag expression, larger study with clinical evaluation and cytogenetic study is recommended to evaluate the impact of those findings over prognosis.

Lactoferrin is an important protein in many biological applications as a potential cancer treatment agent .In this study, lactoferrin was purified from goat colostrum by Ion exchange chromatography by using CM-Sephadex G-150 column and gelfiltration by using Sephadex G-200 column. To evaluted its ability as anticancer agent the study utilized an in vitro evaluation for the cytotoxic effect of the purified goat milk lactoferrin (gLf) on two cell lines, HeLa (Human cervical cancer) cell line and Rat Embryo Fibroblast (REF) cell line at different concentrations and different exposure time of treatment. The purified gLf concentrations ranging (19.53 to 5000) μg/ml for 24,48 and 72 hours. The effect of gLf was evaluated by employing MTT assay. The results revealed significant cytotoxic effect at levels (P<0.05) for all concentrations and for all exposure time of gLF on HeLa cell line as compared to untreated control cells, The inhibition rate IR% increased with raising of gLF concentration and incubation period The highest inhibitory growth was at the concentration ( 5000 μg/ml) after 72hrs of exposure time (64.38%), while only the highest concentration gave significant inhibitory effect (P< 0.05) with normal cell REF.

Objectives: The objectives of present study were to investigate the potential use of mRNA of P53, KRAS, and human telomerase genes in circulating tumor cells as biomarkers for early detection of sporadic cases of colorectal cancer.
Background: Inspite of colorectal cancer is a curable disease if detected early; it is still the third leading cause of death from cancer. There is an ultimate need for implementing an accurate and noninvasive screening method. The recent advances in molecular genetics have showed that the genes of P53, KRAS and human telomerase play major roles in development of colorectal cancer. Therefore the study of their expression in blood samples might be used as a mean for early detection of sporadic colorectal carcinoma cases.
Methods: A case control pilot study in which 20 patients with non-metastatic colorectal carcinoma in Duke’s stages (A – C, TNM from 0 – III) and 20 healthy controls were enrolled. Expression of studied genes and the housekeeping gene Glyceraldehyde-3 phosphate dehydrogenase were determined using quantitative reverse transcriptase polymerase chain reaction. The study was performed in duplicate taking 1μg of total RNA from each sample.
Results: The results showed significant overexpression of mRNAs of studies genes in samples taken from cases of proved colorectal carcinoma compared to that of controls group with a statistically significant P- value of <0.001.
Conclusion: From the above results, the expression of P53, KRAS, and human telomerase genes may be used as biomarkers in the early detection of sporadic non-metastatic colorectal cancer.

This study was conducted to determine the impact of the most important active ingredients which are present in leaves of mint plant. The volatile oil was extracted from the leaves of a mint plant and investigated its toxic effects on the growth of tumor cell lines of cervical cancer( Hela) line human larynx cancer (Hep-2), line of cancer cells the pelvic area RD and compared with a Rat embryonic fibroblast line (REF) ,by using Technology transplant tissue in the glass. The results revealed that significant differences of extract on the tumor cell line , the cytotoxicity for the mentioned line was dependent on concentrations and exposure time 48hr ,while had no effect on the REF normal cells even after 72hr at exposure.

This study is synthesis involved the of amino complex of the silver ion (I), and characterization, in molecular formula of the complex :[Ag(CMZ) NO3.H2O] that stands for CMZ-Carbamazepine .The prepared complex is identified by means of the infrared spectrum (FT-IR) and the Ultraviolet-visible spectrum UV-Vis, the atomic absorption, the accurate elementary analysis (C.H.N), the molar electric conduct ability as well as the measurement of the melting degree. The study showed that the engineering configuration of all prepared complex is tetra-hedral. The effect of the complex is studied (In Vitro) on the Murine mammary adenocarcinoma Cell Line AMN-3. The decimal concentrations are begun at (1,10,100,1000,10000)μg/ml. So it is noticed there is no inhibition on the concentration (1,10,100)μg/ml. The ratio of inhibition is less than 50%. The study is concluded that there is a lethal toxical effect of the complex at the approved concentrations: (1000,10000)μg/ml. The statistical horizontal comparisons among the concentrations of the complex exposure times showed that the 24 hours, exposure to the concentrations(1000,10000)μg/ml. caused significantly affect (56,57.9% respectively) when compared with their corresponding concentrations of other exposure times.

In a study aims to detect the cytogenetic effects of radiation on sample of southern Iraqi peoples have been exposed to during the war in 1991, by using cytogenetic tests on the lymphocytes of blood of the peoples exposed to the bombing. The study has been experimented on 50 randomly samples of exposed peoples .The result show three cases of numerical chromosomal changes ( 6%) and five cases of structural chromosomal changes ( 10%) including inversion, dicentric chromosome, translocation and Philadelphia chromosome. No any abnormal case was defined in control group.

Lactobacillus plantarum bacteria was isolated from different food samples included banana, tomato, mandarin orange, and imported yogurt. Twenty three isoletes of Lactobacillus plantarum were obtained, and identified depending on selective medium MRS agar as well as phenotypic tests, microscopic tests and biochemical examinations. Antioxidant activity of crude bacterial isolates were tested . Results showed that the highest activity was belong to Lactobacillus plantarum isolated from imported yogurt (yR1) 47.12%, while the lower activity was belong to Lactobacillus plantarum isolated from mandarin orange (Mr3) 14.61%. compared with the industrial antioxidant (BHT) and α- tocopherol with activites of antioxidant was (82.70 and 73.96) % respectively. The cytotoxicity effect of the Heat killed (HK) of Lactobacillus plantarum on two of the cancer cells lines, human cervix uteri epitheloid carcinoma (Hela) and rat embryogenic fibroblast (REF) as normal cell line , were studied. In this study, six concentrations of (HK) cells which were (93.5, 187.5, 375, 750,1500, 3000) mcg/ml. The results showed that all types of dead cells of lactobacillus plantarum which clear effect on tumor cells differed depending on the degree of influence For the (HK) cells, the highest percentage of inhibition for imported yogurt was 86.38 after 24 hours. While there was no effect of HK on (Rat Embroy Fibroblast) ofter exposure time 72h.

This study highlights on drawing the attention about the psychological, sociological and ethical problem of persons who
show morphological sex pattern that differs from the true biological gender which is known as intersex. Such cases are born with specific sex and then after short or long period they are discovered with different sex. Most of these cases are not discovered early in order to medically manage but they remain in their apparent sex according to the view and impression of the society for them. Because lack of health education and because the parents do not pay high attention to the external genitals of these cases, the parents do not visit the doctor to seek treatment for them. Therefore these cases grow according to the apparent sex that the society decision. Within the year medically, psychological and social problems start to appear that require medical intervention, and most these cases are discovered during or after puberty. Discover the true gender for such cases will raise big medically, ethical, psychological and social problems that urgently need to be studied and analysed carefully to find acceptable scientific approaches to offer support for such cases to help them to adapt to the new conditions and to accept their new gender with courage. Therefore, this study aims to draw attention to the problems faced by intersex cases and how they suffer from future challenges that require solutions in the short and long term. This study revealed that 243 intersex cases during the last two years for both sexes. The adult intersex cases showed sever great suffering from they were before they discover their real sex, as well as showed the biggest fear and suffering after the discovery of their real sex. Their parents and relatives showed obvious and large concerns after the discovery of real sex to those cases and remained loss and embarrassed to how to cope with the shock and how to face of the community. The study recommends great attention to those cases and provides medical, psychological and social support for them to help them to overcome the transitional period that they are going through after the discovery of the new gender, especially in Iraq, because of restricted local customs, habits, religious and traditions.