Targeting PERK and GRP78 in colorectal cancer: Genetic insights and novel therapeutic approaches.

Colorectal cancer (CRC) ranks among the leading causes of cancer-related deaths worldwide. Enhancing CRC diagnosis and prognosis requires the development of improved biomarkers and therapeutic targets. Emerging evidence suggests that the unfolded protein response (UPR) plays a pivotal role in CRC progression, presenting new opportunities for diagnosis, treatment, and prevention. This study hypothesizes that genetic variants in endoplasmic reticulum (ER) stress response genes influence CRC susceptibility. We examined the frequencies of SNPs in PERK (rs13045) and GRP78/BiP (rs430397) within a South Iranian cohort. We mapped the cellular and molecular features of PERK and GRP78 genes in colorectal cancer, observing their differential expressions in tumor and metastatic tissues. We constructed co-expression and protein-protein interaction networks and performed gene set enrichment analysis, highlighting autophagy as a significant pathway through KEGG. Furthermore, the study included 64 CRC patients and 60 control subjects. DNA extraction and genotyping were conducted using high-resolution melting (HRM) analysis. Significant differences in PERK and GRP78 expressions were observed between CRC tissues and controls. Variations in PERK and GRP78 genotypes were significantly correlated with CRC risk. Utilizing a Multi-Target Directed Ligands approach, a dual PERK/GRP78 inhibitor was designed and subjected to molecular modeling studies. Docking experiments indicated high-affinity binding between the proposed inhibitor and both genes, PERK and GRP78, suggesting a novel therapy for CRC. These findings highlight the importance of understanding genetic backgrounds in different populations to assess CRC risk. Polymorphisms in UPR signaling pathway elements may serve as potential markers for predicting CRC susceptibility, paving the way for personalized therapeutic strategies.

Multi-Drug Resistance against Second-Line Medication and MicroRNA Plasma Level in Metastatic Breast Cancer Patients.

Background: Circulating microRNAs (miRNAs) can help to predict the chemotherapy response in breast cancer with promising results. The aim of the present study was to investigate the relationships between the miR-199a, miR-663a, and miR-663b expression and chemotherapy response in metastatic breast cancer patients. Methods: This study is a case-control study performed at Yasuj University of Medical Sciences (2018-2021). The expression levels of miR-663a, miR-663b, and miR-199a in the serum of 25 patients with metastatic breast cancer versus 15 healthy individuals were determined by the real-time polymerase chain reaction method. The response to treatment was followed up in a 24-month period. All patients were treated with second-line medications. Two or more combinations of these drugs were used: gemcitabine, Navelbine®, Diphereline®, Xeloda®, letrozole, Aromasin®, and Zolena®. Statistical analyses were performed in SPSS 21.0 and GraphPad Prism 6 software. The expression levels were presented as mean±SD and analyzed by Student's t test. Results: The results and clinicopathological features of patients were analyzed by t test. The statistical analysis showed that miR-663a expression was related to human epidermal growth factor receptor 2 (HER2) status and was significantly lower in the HER2+ than HER2- group (P=0.027). Moreover, the expression of miR-199a and miR-663b was significantly correlated with the response to treatment, in which the expression of miR-199a was higher in the poor-response group (P=0.049), while the higher expression of miR-663b was seen in the good-response group (P=0.009). Conclusion: These findings state that the high plasma level of miR-199a and the low plasma level of miR-663b may be related to chemoresistance in patients with metastatic breast cancer.

Circulating mRNA and plasma levels of osteoprotegerin and receptor activator of NF-κB ligand in nonalcoholic fatty liver disease.

Pathogenesis of the beginning and progression of nonalcoholic fatty liver disease (NAFLD) has not been clarified exactly. The osteoprotegerin (OPG)/receptor activator of NF-κB ligand (RANKL) axis seems to play an imperative function in the onset and progression of this disease. The goal of the present study was to investigate the peripheral blood mononuclear cell (PBMC) expression and plasma levels of RANKL and OPG cytokines in NAFLD patients and compare them with healthy group. Plasma levels of OPG and RANKL were determined with ELISA kits in 57 men with NAFLD and 25 healthy men as controls. Biochemical and anthropometric parameters tests were also evaluated in the study groups. RANKL and OPG mRNA contents were evaluated by quantitative RT-PCR. OPG contents were markedly decreased in NAFLD patients as compared with healthy patients [1.43 (1.05-5.45)] versus [2.94 (1.76-4.73)] ng/mL; P = 0.007). The levels of RANKL were significantly reduced in NAFLD patients [74.00 (56.26-203.52) ng/mL] than in healthy patients [119.37 (83.71-150.13) ng/mL]; (P = 0.03). Also, OPG and RANKL gene expression were significantly decreased in NAFLD patients in comparison with the control group (P < 0.05). Moreover, receiver operating characteristic curve indicated that OPG may have a good capability to discriminate between NAFLD patients and normal individuals. A positive correlation was observed between OPG and RANKL in plasma sample (r = 0.495) (P = 0.000). Decreased plasma levels and gene expression of RANKL and OPG cytokines in NAFLD patients indicate that there is a relationship between these cytokines and the pathology of NAFLD disease. Confirmation of this association as well as the mechanism and role of these cytokines in NAFLD require further studies.

Intercalation of curcumin into liposomal chemotherapeutic agent augments apoptosis in breast cancer cells.

Resistance to common chemotherapeutic agents is a frequent phenomenon in late-stage breast cancers. An ideal system capable of the co-delivery of hydrophobic and hydrophilic chemotherapeutic agents can regulate the dosage and co-localization of pharmaceutical compounds and thereby improve the anticancer efficacy. Here, for the first time, we have intercalated curcumin (Cur) into a double-layered membrane of cisplatin (Cis) liposomes to obtain a dosage controlled co-delivery formulation, capable of inducing apoptosis in breast cancer cells. The concentrations of Cur and Cis in nanoliposome (Cur-Cis@NLP) were optimized by response surface methodology (RSM); RSM optimization showed 99.81 and 23.86% entrapment efficiency for Cur and Cis, respectively. TEM analysis demonstrated the fabrication of nanoparticles with average diameter of 100 nm. The anticancer and apoptotic effects of Cur-Cis@NLPs were also evaluated using MTT assay, fluorescent staining and flow cytometry assays. Cytotoxicity assessments of various Cur-Cis@NLPs concentrations demonstrated a concentration-dependent manner. In comparison to free and liposomal Cis, Cur-Cis@NLP reduced breast cancer cells' viability (82.5%) in a significant manner at a final concentration of 32 µg.mL-1 and 20 µg.mL-1 of Cur and Cis, respectively. Combination index values calculation of Cur-Cis@NLP showed an overall CI value <1, indicating synergetic effect of the designed co-delivery system. Additionally, flow cytometry assay demonstrated Cur-Cis@NLPs triggered apoptosis about 10-folds higher than liposomal Cis. This co-drug delivery system has a potential for the encapsulation and release of both hydrophobic and hydrophilic drugs, while taking the advantages of the reduced cytotoxic effect along with achieving high potency.

Association of single nucleotide autophagy-related protein 5 gene polymorphism rs2245214 with susceptibility to non-small cell lung cancer.

INTRODUCTION: Autophagy is a mechanism that is involved in the regulation of cellular life, apoptosis, and stemness while its intervening genes play important functions in various cancers including lung cancer. ATG5 is one of the key genes for the regulation of the autophagy pathway. In this study, our team has investigated the potential relationship between ATG5 gene polymorphism rs2245214 with non-small cell lung cancer (NSCLC) in a subpopulation of patients from southern Iran. In this study, 34 patients with NSCLC (20 males and 14 females [mean age: 12.86 ± 60.47 years]) and 50 healthy subjects (30 males and 20 females [mean age: 13.09 ± 56.62 years]) were studied in terms of the genotype of the ATG5 gene. We used restriction fragment length polymorphism and analyzed the results using SPSS software (v.23). The results revealed that subjects harboring the guanine/cytosine (GC) genotype of the rs2245214 ATG5 gene polymorphism had suffered less from NSCLC, whereas the prevalence of the C-allele of this polymorphism was significantly higher in patients with NSCLC ( P < 0.05). On the basis of the results of logistic regression, the presence of this C-allele may predict the risk of lung cancer ( P value = 0.011; OR, 3.52; 95% CI, 1.33-9.26). This study concludes that the C-allele of the rs2245214 ATG5 gene polymorphism is associated with increased susceptibility to NSCLC, whereas the GC genotype of this polymorphism is associated with decreased risk and might therefore have a protective role in the development of NSCLC.

Relationship of SNP rs2645429 in Farnesyl-Diphosphate Farnesyltransferase 1 Gene Promoter with Susceptibility to Lung Cancer.

BACKGROUND AND PURPOSE: The mevalonate pathway is one of the major metabolic pathways that use acetyl-CoA to produce sterols and isoprenoids. These compounds can be effective in the growth and development of tumors. One of the enzymes involved in the mevalonate pathway is FDFT1. Different variants of this gene are involved in the risk of suffering various diseases. The present study examined the relationship between FDFT1 rs2645429 polymorphism and the risk of nonsmall cell lung cancer (NSCLC) in a population from southern Iran. METHOD: The genotypes of rs2645429 polymorphism of FDFT1 gene were examined in 95 samples: 34 patients with NSCLC and 61 healthy individuals by RFLP method. RESULTS: The results of this study indicated that C allele of this polymorphism was effectively associated with the risk of NSCLC in the Iranian population (p value = 0.023; OR = 2.71; 95% CI = 1.12-6.59) and CC genotype has significant relation with susceptibility to NSCLC (p value = 0.029; OR = 3.02; 95% CI = 1.09-8.39). This polymorphism is located in the promoter region FDFT1 gene, and CC genotype may increase the activity of this promoter. This study also found a significant relationship between C allele and metastatic status. C allele was more common in NSCLC patients. (p = 0.04). CONCLUSION: C allele of FDFT1 rs2645429 polymorphism gene can be a risk factor for NSCLC, whereas T allele probably has a low protective role.

Using curcumin to prevent structural impairments of testicles in rats induced by sodium metabisulfite.

Sodium metabisulfite (Na-MBS) is a disinfectant and preservative agent. Some organ including testicle would be in danger in the case of Na-MBS consumption. Curcumin (CUR) is the constituent of turmeric with protective properties. The effect of CUR on testicles in rats exposed to Na-MBS evaluated using stereological methods. Sprague-Dawley rats were divided into eight groups. The rats in groups I to VIII received the following respectively: distilled water, CUR (100 mg/kg/day), low (0.7 mg/kg/day: acceptable daily intake), intermediate (7 mg/ kg/day), and high (70 mg/kg/day) doses of Na-MBS, and low, intermediate, and high doses of Na-MBS plus CUR. After 7 weeks, the testicles were analyzed. The volume of seminiferous tubule, tubular epithelium and tubule length reduced (25-40 %) on average in the rats that received intermediate and high doses of Na-MBS, while the connective tissue volume increased (15-20 %) in both groups (P<0.01). Besides, 19-36 % and 41-57 % of the cells (spermatogonia types A and B, spermatids, Sertoli and Leydig) were lost in the rats that received intermediate and high doses of Na-MBS respectively in comparison to the control groups. Nonetheless, all the above-mentioned alterations ameliorated drastically in the rats that received Na-MBS plus CUR compared to those exposed to Na-MBS without CUR therapy (P<0.01). The acceptable daily intake of Na-MBS for 7 weeks did not affect on testicular parameters. CUR (100 mg/kg/day) could prevent structural impairments of testicles in the rats induced by Na-MBS (7 and 70 mg/kg/day).

Effect of Antioxidants (ß-mercaptoethanol and Cysteamine) on Assisted Reproductive Technology In vitro.

INTRODUCTION: Oocyte Culture of Germinal Vesicle (GV) and its growth improves Assisted Reproductive Technology (ART) invitro and infertility. Inappropriate culture medium environment, low quality of oocytes, increase in Oxidative Stress (OS) events, Reactive Oxygen Species (ROS) and free radicals production are the main factors that result in unsuccessful Invitro Maturation (IVM) and decrease in reproduction. AIM: The present study was conducted with the aim to evaluate the effect of ß-mercaptoethanol (BME) and Cysteamine (CYS) on IVM improvement, embryo fertilization and development of blastocyst of mouse immature oocyte. MATERIALS AND METHODS: Oocytes were obtained from 4-6 weeks old Naval Medical Research Institute (NMRI) female mice, 48 hours after stimulation with Intraperitoneal (IP) injection of 10 IU Pregnant Mare Serum Gonadotropin (PMSG). GV oocyte with and without cumulus cells were isolated from ovaries and cultured in Tissue Culture Medium (TCM) 199 with availability of 100 µM of antioxidants (BME and CYS). After 24 hours, mature oocyte in metaphase II (MII) were fertilized with sperm in In vitro Fertilization (IVF) medium (T6) and evaluated for fetal development into blastocyst. RESULTS: BME and CYS could significantly (p<0.05) increase the rate of IVM and oocyte evolution, and embryo formation in medium culture. Furthermore, it is demonstrated that existence of Cumulus Oocyte Complexes (COC) significantly showed better IVM, fertilization and evolution trend as compared to oocytes without cumulus cover or Denuded Oocytes (DO), especially in TCM199 plus BME and CYS. So that the change in GV stage oocytes to MII (maturation rate), fertilization rates or 2PN formation, and two cell embryos formation or blastocyst development rate in the treatment group with addition of BME & CYS and COC was statistically significant as compared to the DO group (p-value < 0.0001). CONCLUSION: Both cellular and environmental factors could be important and involved in ART improvement.

The Hydroalcoholic Extract of Matricaria chamomilla Suppresses Migration and Invasion of Human Breast Cancer MDA-MB-468 and MCF-7 Cell Lines.

BACKGROUND: Matricaria chamomilla is an aromatic plant with antioxidant, anticancer, and anti-inflammatory properties. However, the inhibitory role of M. chamomilla on migration and invasion of human breast cancer cells remains unclear. OBJECTIVE: This study investigated the methods to evaluate these anticancer mechanisms of M. chamomilla on human breast cancer MCF-7 and MDA-MB-468 cell lines. MATERIALS AND METHODS: The cells were treated with hydroalcoholic extract of M. chamomilla at different concentrations (50-1300 µg/mL) for 24, 48, and 72 h in a culture medium containing 10% fetal bovine serum. This study quantified the 50% growth inhibition concentrations (IC50) by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay; apoptosis and necrosis through Hoechst 33342/propidium iodide staining; cell proliferation and clone formation by clonogenic assay as well as cellular migration, invasion, and attachment. After 24, 48, and 72 h of treatment, the IC50levels were 992 ± 2.3 µg/mL, 893 ± 5.4 µg/mL, and 785 ± 4.8 µg/mL against MDA-MB-468, respectively, and 1288 ± 5.6 µg/mL, 926 ± 2.5 µg/mL, and 921 ± 3.5 µg/mL, against MCF-7, respectively. Furthermore, increasing the extract concentrations induced cellular apoptosis and necrosis and decreased cellular invasion or migration through 8 µm pores, colonization and attachment in a dose-dependent manner. RESULTS: It indicated time- and dose-dependent anti-invasive and antimigrative or proliferative and antitoxic effects of hydroalcoholic extract of aerial parts of chamomile on breast cancer cells. CONCLUSION: This study demonstrated an effective plant in preventing or treating breast cancer. SUMMARY: Antioxidant compounds in Matricaria chamomilla have anticancer effects.Hydroalcoholic extract of M. chamomilla controls cellular proliferation and apoptosis induction.Hoechst 33342/propidium iodide staining suggested that the extract induces apoptosis more than necrosis.Hydroalcoholic extract of M. chamomilla prevents colonization and cellular migration of human breast cancer MDA-MB-468 and MCF-7 cell lines in a time- and dose-dependent manner.M. chamomilla has low cytotoxic effects on natural cells. Abbreviations Used: IARC: International Agency for Research on Cancer; WHO: World Health Organization; FBS: Fetal bovine serum; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; DMSO: Dimethyl sulfoxide; PI: Propidium iodide; LN: Live cells with normal nucleus; LA: Live cells with apoptized nucleus; DN: Dead cells with normal nucleus; DA: Dead cells with apoptized nucleus; BSA: Bovine serum albumin; ANOVA: Analysis of variance; IC50: 50% growth inhibition concentration; GSE: Grape seed extract.

Polymorphisms of Leptin (-2548 G/A) and Leptin Receptor (Q223R) Genes in Iranian Women with Breast Cancer.

Recent studies have shown that polymorphisms in leptin and leptin receptor genes are associated with increased risk for breast cancer. This study aimed at investigating -2548 G/A polymorphism in leptin gene and Q223R polymorphism in leptin receptor gene in patients with breast cancer. The study included 45 women with breast cancer and 41 healthy women. PCR-RFLP was used to determine the genotype of the subjects in terms of -2548 G/A polymorphism in leptin gene and Q223R polymorphism in leptin receptor gene. Serum levels of leptin were also measured by ELISA. For -2548 G/A polymorphism, the genotypes were homozygous AA (OR = 1.13; p = 0.8) and heterozygous GA (OR = 0.41; p = 0.2) and for Q223R polymorphism, the genotypes were homozygous RR (OR = 6.7; p = 0.09) and heterozygous QR (OR = 8.3; p = 0.06). The mean serum level of leptin was 33.22 ± 21.35 ng/mL in patients and 29.49 ± 23.27 ng/mL in the normal participants (p = 0.3). Although, despite the magnitude of the associations, the results suggested no statistically significant contribution of -2548 G/A polymorphism (in leptin gene), Q223R polymorphism (in leptin receptor gene), and serum leptin levels in predicting the risk of breast cancer, further studies with larger sample size are suggested.

The Nuclear Maturation and Embryo Development of Mice Germinal Vesicle Oocytes with and without Cumulus Cell after Vitrification.

BACKGROUND: Cryobiology is an essential tool in assisted reproductive technology. Research in this area focuses on the possibility of restoring fertility in women with reproductive problems or after cancer treatments. AIM: The purpose of this study was to evaluate viability of oocytes, In vitro maturation and embryo development in vitrified germinal vesicle oocytes with and without cumulus cell after single and stepwise vitrification procedure. MATERIALS AND METHODS: Germinal vesicle oocytes with or without cumulus cells were obtained from 4 weeks old female mice 48h after intraperitoneal injection of 7.5 IU pregnant mare serum gonadotropin (PMSG). For vitrification collected oocytes vitrification were exposed to cryoprotectant, which was composed of 30% (v/v) ethylene glycol, 18% (w/v) Ficoll-70, and 0.3 M sucrose, either by single step or in a step-wise way. After exposure to cryoprotectant and immerged in liquid nitrogen, the oocytes were thawed and washed in medium TCM199 two times. Then the oocytes transferred to IVM medium for maturation and embryo development to blastocyst. RESULTS: The oocytes survival rates after vitrifying-warming, maturation rate, the capacity of fertilization and embryonic development to blastocyst were examined in vitro. The oocytes survival, maturation to MII, fertilization developmental rate in the step-wise exposure and with cumulus cell was significantly higher (p<0.05) as compared with corresponding rate in the single step procedure without cumulus cell. CONCLUSION: The results of present study indicated that oocytes vitrified with cumulus cells and stepwise procedure had positive effect on maturation and developmental rate to blastocyst than oocytes without cumulus cell and single step procedure.

Effects of Pomegranate Seed Oil on the Fertilization Potency of Rat's Sperm.

BACKGROUND: Pomegranate has been taken great scientific attention in recent years due to its health benefits. Pomegranate seed oil is a rich source of 9-cis, and 11-trans conjugate linolenic acid. The aim of this study was to evaluate the effect of dietary pomegranate seed oil on the fertilization potency of rat's sperm. MATERIALS AND METHODS: Twenty-four male Wistar rats were divided into four groups. The first group, which served as the control group, received 1 mL of corn oil for seven weeks. Groups II, III, IV served as the experimental groups received 200, 500 and 1000 mg/kg of pomegranate seed oil, for the same period of time respectively. After seven weeks, all of the rats were sacrificed, and their epididymis sperm was collected and added to IVF medium (T6) containing metaphase II oocytes. Almost 21 oocytes had been removed from every female rat oviduct. In this medium, oocyte fertilization, cleavage rates, and embryo development into blastocysts, were evaluated by inverted microscopy. RESULTS: Levels of LD50 in the oral route in male rats were more than 5000 mg/kg body weight. Our data showed that the rates of fertilization, cleavage and embryo development into blastocysts were higher in the groups that had received 500 and 1000 mg/kg body weight of pomegranate seed oil. CONCLUSION: This study demonstrated that pomegranate seed oil had a positive effect on the fertilization potency of male rats. These beneficial effects may be useful in assisted reproductive technology.

Quantitative Analysis of ErbB1 and ErbB2 Genes Amplification by a High Performance Liquid Chromatography.

BACKGROUND: Genes for human epidermal growth factor receptors B1 (ErbB1) and B2 (ErbB2) were amplified in breast and ovarian cancers. Both of them were associated with aggressive disease and worse prognosis. The ErbB1 or ErbB2 status of a tumor may provide an indication of the response to ErbB1 and ErbB2 -targeted therapies. For accurate and rapid assessment of amplification of ErbB1 and ErbB2 oncogenes, a High Performance Liquid Chromatography (HPLC) method was developed in this study. METHODS: DNA was extracted from 30 primary breast tumors and 20 blood samples of healthy donors. ErbB1 and ErbB2 genes along with a reference gene were co-amplificated by Polymerase Chain Reaction (PCR). The PCR products were separated and quantified using an anion-exchange column within 30 min and in a single step. Optimum resolution was obtained when a sodium chloride gradient and a column temperature of 35°C were used. The results of HPLC analysis of ErbB1 and ErbB2 PCR products were compared with real time PCR method as a gold standard test for 7 tumor samples. RESULTS: The proposed HPLC method was confirmed by real time PCR method. Twenty two and ten of the specimens in our breast cancer cohort showed more than a two-fold amplification of ErbB2 and ErbB1 oncogenes, respectively. CONCLUSION: Our results were confirmed by real time PCR and showed that HPLC method is a specific, cheap and clinically applicable analytical approach for assessment of ErbB1 and ErbB2 statuses in breast tumors.

UBE2Q1, as a Down Regulated Gene in Pediatric Acute Lymphoblastic Leukemia.

Ubiquitin - proteasome system (UPS), the major protein degradation pathway in the cells, typically degrades short - lived and damaged proteins and regulates growth and stress responses. This pathway is altered in various cancers, including Acute Lymphoblastic Leukemia (ALL). ALL begins with a change in bone marrow cells and is the most common type of leukemia in children under 15 years. UBE2Q1 as a new characterized gene of E2 enzyme family is located on chromosome 1 and reported to be altered in some malignancies. In this study, we aimed to explore the expression pattern of UBE2Q1 gene in children with ALL. For this purpose, a series of RT - PCR and quantitative RT - PCR were performed on a collection of 20 bone marrow samples of ALL patients and the same number of whole blood samples of age - matched normal subjects. Gel electrophoresis of RT - PCR products revealed the expression of UBE2Q1 mRNA in most of the normal (90%) and about half of the leukemic (45%) samples. QRT - PCR data indicated that only 1 patient out of 20 (5%) showed up regulation of the gene (> 2 folds). In 4 patients (20%), the expression of UBE2Q1 mRNA was equivocal (from 1/2 to 2) and in 15 cases (75%), the gene was down regulated (> 1/2) when compared to the normal samples. In conclusion, down regulation of UBE2Q1 in the majority of the leukemic samples suggests its potential implication in the pathogenesis of ALL. UBE2Q1 can be considered as a molecular marker and a candidate targeting to treat ALL in the future.

Expression Status of UBE2Q2 in Colorectal Primary Tumors and Cell Lines.

BACKGROUND: Activation of the ubiquitin-proteasome pathway in various malignancies, including colorectal cancer, is established. This pathway mediates the degradation of damaged proteins and regulates growth and stress response. The novel human gene, UBE2Q2, with a putative ubiquitin-conjugating enzyme activity, is reported to be overexpressed in some malignancies. We sought to investigate the expression levels of the UBE2Q2 gene in colorectal cell lines as well as in cancerous and normal tissues from patients with colorectal cancer. METHODS: Levels of UBE2Q2 mRNA in cell lines were assessed by Real-Time PCR. Western blotting was employed to investigate the levels of the UBE2Q2 protein in 8 colorectal cell lines and 43 colorectal tumor samples. RESULTS: Expression of UBE2Q2 was observed at the level of both mRNA and protein in colorectal cell lines, HT29/219, LS180, SW742, Caco2, HTC116, SW48, SW480, and SW1116. Increased levels of UBE2Q2 immunoreactivity was observed in the 65.11% (28 out of 43) of the colorectal carcinoma tissues when compared with their corresponding normal tissues. Difference between the mean intensities of UBE2Q2 bands from cancerous and normal tissues was statistically significant at P<0.001 (paired t test). CONCLUSION: We showed the expression pattern of the novel human gene, UBE2Q2, in 8 colorectal cell lines. Overexpression of UBE2Q2 in the majority of the colorectal carcinoma samples denotes that it may have implications for the pathogenesis of colorectal cancer.

Influence of Insulin-Like Growth Factor-I on Maturation and Fertilization Rate of Immature Oocyte and Embryo Development in NMRI Mouse with TCM199 and α-MEM Medium.

INTRODUCTION: In vitro maturation (IVM) of oocytes and subsequent, in vitro fertilization (IVF) for the generation of embryos in the laboratory has important values. Growth factors are a component of a complex system of autocrine and paracrine factors that have a regulatory role in ovarian function and affect oocyte maturation. Therefore, the aim of this study is to evaluate the effect of IGF-I on IVM and IVF of mice oocytes during culture with α-MEM and TCM199 medium. MATERIALS AND METHODS: Cumulus oocyte complexes (COCs) and denuded oocyte were obtained from 4-6 week old NMRI mice and underwent in vitro maturation and in vitro fertilization in presence or absence of IGF-I with α-MEM and TCM199. RESULT: Maturation rate (79.6%), fertilization rate (87.2%), two cells development rate (79.5%) and blastocyst rate(43.2%) was higher in COCs cultured in α-MEM with IGF-I, while lower maturation rate (50.6%) fertilization rate (61%), two cells development rate (48.8%) and blastocyst rate(14.6%) were seen in cultured denuded oocytes (DOs) in TCM199 without growth factor. As well as, maturation fertilization, two cells development and blastocyst rates in COCs were higher than DOs. CONCLUSION: Our findings have shown that IGF-I is involved in the oocyte biology and improve the oocyte maturation, fertilization and embryo development to blastocyst competence in vitro. In addition, it has also shown that cumulus cells are vital for oocyte development when IGF-1 added to the mediums.

UBE2Q1 expression in human colorectal tumors and cell lines.

Colorectal cancer is the third most common cancer in the world. Ubiquitin-proteasome system has shown to be activated in colorectal and other malignancies. UBE2Q1 is a novel human gene that encodes a putative E2 ubiquitin conjugating enzyme. Here, we investigated the expression pattern of UBE2Q1 gene in cell lines and tissues from human colorectal tumors. Quantitative (q) RT-PCR were employed to evaluate the expression levels of the mRNA for UBE2Q1 in colorectal cancer cell lines (HT29/219, LS180, SW742, Caco2, HTC116, SW48, SW480 and SW1116). Expression of UBE2Q1 at the protein levels were assessed by Western blotting in cell lines as well as in 43 human colorectal tumor tissues. All cell lines tested expressed UBE2Q1 gene at the level of both mRNA and protein, with the SW1116 line representing the lowest level of expression. The cell lines HT29/219 and SW742 showed the highest levels of UBE2Q1 protein and mRNA respectively. When compared to corresponding normal tissues, malignant parts of colorectal tumors showed increased levels of UBE2Q1 immunoreactivity in 32 (74.42 %) of cases. These data suggest that UBE2Q1 is differentially expressed in colorectal cell lines and shows overexpression in colorectal tumors.

Expression of UBE2Q2, a putative member of the ubiquitin-conjugating enzyme family in pediatric acute lymphoblastic leukemia.

BACKGROUND: Acute lymphoblastic leukemia (ALL) is a cancer of the white blood cells most commonly found in childhood with a peak incidence at 2-5 years of age. The ubiquitin degradation pathway facilitates degradation of damaged proteins and regulates the growth and stress response. This pathway is activated in various cancers, including ALL. It has been previously reported that the newly characterized human gene UBE2Q2, a putative member of the ubiquitin-conjugating enzyme family, is over-expressed in the tumor mass and invasive epithelium in head and neck squamous cell carcinoma and breast cancer. METHODS: Here, we have used quantitative reverse transcriptase polymerase chain reaction (RT-PCR) to assess expression of the UBE2Q2 gene in bone marrow samples of 20 children with ALL. Whole blood samples of 20 normal children were used as control specimens.  RESULTS: RT-PCR revealed the expression of UBE2Q2 mRNA in 80% of the bone marrow samples from ALL patients as well as in 85% of leukemic normal peripheral blood cells. According to the results of quantitative RT-PCR, the levels of UBE2Q2 mRNA expression in the bone marrow cells of 11 out of the 20 children with ALL (55%) were significantly higher (> 2-47 fold) than those in blood cells of normal children. CONCLUSION: Our data suggest that the newly characterized human gene, UBE2Q2, may have implications for the pathogenesis of ALL and could be used for molecular diagnosis purposes in the future.

Expression of the novel human gene, UBE2Q1, in breast tumors.

The novel human gene, designated ubiquitin-conjugating enzyme E2Q family member 1 (UBE2Q1) maps to chromosome 1q21.3. The gene has an open reading frame corresponding to 422 amino acids and contains a RWD domain and an E2 ubiquitin conjugating enzyme domain. Here, we investigated the expression levels of both mRNA and protein of UBE2Q1 gene in cancerous versus normal parts of breast specimens from 26 patients. Real-time PCR data showed that the relative expression level of UBE2Q1 mRNA was significantly greater in cancers than in non-cancerous tissues of breast specimens (Mean ± SEM, 0.064 ± 0.015 for cancers and 0.026 ± 0.01 for noncancerous tissues, P < 0.05 Mann-Whitney test). A rabbit polyclonal antibody was generated against an amino acid sequence predicted from the DNA sequence of UBE2Q1 gene. This antibody was used to perform Western blotting on 21 cases in our cohort of breast specimens. Thus, 13 (61.904%) of the cases showed an increase in the UBE2Q1 immunoreactivity in their cancerous tissues as compared with the corresponding normal tissues. This result along with the real-time PCR data shows that the novel human gene, UBE2Q1, is expressed in human breast and may have implications for pathogenesis of breast cancer.

Effect of oral resveratrol on the BDNF gene expression in the hippocampus of the rat brain.

Resveratrol is a plant polyphenolic compound. Evidence indicates that resveratrol has beneficial effects against aging and neurodegenerative diseases. The goal of our study was in vivo examination of the effects of resveratrol on the abundance of mRNA encoding Brain Derived Neurotrophic Factor (BDNF) in the hippocampus of rat brain. Rats were administrated orally by different doses (2.5-20 mg/kg bwt) of resveratrol for 3, 10 and 30 days. Saline was used as control and 10% ethanol in saline was used as vehicle for resveratrol. Measurement of BDNF mRNA by Real-time RT-PCR showed that levels of the mRNA for BDNF were significantly and dose dependently elevated in the hippocampal tissues of rats. The findings suggest that the neuroprotective effects of resveratrol may be at least partly due to its inducing effects on the expression levels of the BDNF mRNA.