Molecular Characterization, Expression and Functional Analysis of Yak IFITM3 Gene

Background IFITM3 is interferon-induced transmembrane 3, which plays an extremely key role in anti-proliferation, anti-virus and anti-tumor diseases. To expand our understanding of the role of IFITM3 in yak, this experiment studied its function. Results Firstly, the yak ( Bos grunniens ) IFITM3 ( BgIFITM3 ) gene contained a 5’-untranslated region (UTR) (25 bp), a coding region (441 bp), and a 3’-UTR (115 bp). The expression of BgIFITM3 gene in liver was significantly higher than that in heart, spleen, lung and kidney ( P <0.01). BgIFITM3 protein was localized on the yak hepatocyte membrane, and its expression level was increased first and then stabilized from 1 day to 5 years of age. Moreover, the prokaryotic expression vector of BgIFITM3 protein was constructed and expressed successfully, with a molecular weight of 19.5 kDa. Besides, the activity of yak hepatocyte was significantly inhibited after treating with BgIFITM3 protein (10 and 20 μg/mL) ( P <0.01). The expression levels of ERBB-2, IRS-1, PI3KR-1, AKT-1 and MAPK-3 were significantly lower after treating with 20 μg/mL BgIFITM3 protein ( P <0.05). Finally, the activity of HepG2 cells was significantly inhibited after treating with BgIFITM3 protein (1, 10 and 20 μg/mL) ( P <0.05). While the cloning ability and migration ability of HepG2 cells were significantly inhibited after treating with 10 μg/mL BgIFITM3 protein ( P <0.05). The mitochondria of HepG2 cells were concentrated, cristae widened, and the double film density of mitochondria was increased after treating with 10 μg/mL BgIFITM3 protein. After 10 μg/mL BgIFITM3 protein treating, the expression levels of VDAC-2, VDAC-3 , p53 genes were significantly increased, but the expression level of GPX-4 gene was significantly decreased ( P <0.01). Conclusion Taken together, the BgIFITM3 protein could inhibit the proliferations of yak hepatocyte and HepG2 cells by regulating the PI3K/Akt pathway or ferroptosis-related genes, respectively. These results benefit for further study of the function of BgIFITM3 protein.

Characterization of the Components and Pharmacological Effects of Mountain-Cultivated Ginseng and Garden Ginseng Based on the Integrative Pharmacology Strategy

Panax ginseng C. A. Mey (PGCAM) is a herbaceous perennial belonging to the Araliaceae family, mainly including Mountain-Cultivated Ginseng (MCG) and Garden Ginseng (GG) on the market. We aimed to establish a rapid, accurate and effective method to distinguish 15-year-old MCG and GG using ultra-performance liquid chromatography-quadrupole time-of-flight-tandem mass spectrometry (UPLC-QTOF-MS/MS), and also explored the pharmacological mechanisms of the main components using the Integrative Pharmacology-based Network Computational Research Platform of Traditional Chinese Medicine (TCMIP V2.0; http://www.tcmip.cn/). Altogether, 23 potential quality markers were characterized to distinguish 15-year-old MCG and GG, including ginsenosides Ra2, Rg1, and Ra1, and malonyl-ginsenoside Ra3, etc. The contents of 19 constituents (mainly protopanaxadiol-type) were higher in MCG compared with that in GG, and four constituents (mainly carbohydrate compounds) were higher in GG. The 105 putative targets corresponding to 23 potential quality markers were mainly involved in 30 pathways, which could be divided into 10 models, such as immune regulation, systems (metabolic, nervous, cardiovascular, reproductive), blood-pressure regulation, as well as antitumor, antiaging, antibacterial and anti-inflammatory effects. Furthermore, the potential quality markers of MCG and GG could inhibit the proliferation of breast cancer by regulating the mRNA expression of PSA, S6K, MDM2, and P53 genes by acting on AR, MTOR, PI3K and other targets. The Integrative Pharmacology Strategy may provide an efficient way to identify chemical constituents and explore the pharmacological actions of TCM formulations.

Target genetic abnormalities for the treatment of colon cancer and its progression to metastasis

: Colorectal carcinogenesis involves various processes from the accumulation of genetic alterations to genetic and epigenetic modulations and chromosomal abnormalities. It also involves mutations in oncogenes and tumour suppressor genes. Genomic instability plays a vital role in CRC. Advances in modern biological techniques and molecular level studies have identified various genes that are involved in colorectal cancer (CRC). KRAS, BRAF, PI3K, and p53 genes play a significant role in different phases of CRC. Alteration of these genes leads to development or progression and metastasis colon cancer. This review focuses on the role of KRAS, BRAF, PI3KCA, and TP53 genes in carcinogenesis and their significance in various stages of CRC. It also provides insights on specific modulators acting on these genes. Further, this review discusses the mechanism of the pathways involving these genes in carcinogenesis and current molecules and treatment options under various stages of clinical evaluation.

P53 and PIK3CA Mutations in KRAS/HER2 Negative Ovarian Intestinal-Type Mucinous Carcinoma Associated with Mature Teratoma

Primary ovarian intestinal-type mucinous carcinomas associated with mature teratoma are rare and represent less than 3% of all primary ovarian neoplasms. The molecular profile of these tumors is still controversial. We report here the first case of mucinous ovarian tumor in which mutation of the PIK3CA and P53 genes could be demonstrated by the next generation sequencing technique without KRAS mutation or HER2 amplification. Our data suggest that these mucinous carcinoma variants probably present an extremely complex molecular biology profile that should be known in the future to stratify therapeutic outcomes and potential targeted therapies, particularly in recurrent disease.

MEASUREMENT OF DNA DAMAGE, OXIDATIVE STRESS, AND GENE EXPRESSION OF β-CATENIN AND P53 GENES IN LIVER AND BRAIN OF MALE MICE RECEIVING MONOSODIUM L-GLUTAMATE MONOHYDRATE

Introduction: Monosodium L-glutamate (MSG) monohydrate is a widespread nutritional additive and flavoring agent frequently consumed all over the world. In this study, we investigate the action of daily oral intake of MSG monohydrate in vivo using mammalian systems. Methods: Mice divided as follows: Group I (normal control), Group II, and Group III treated with MSG for 2 and 4 weeks, respectively. Brain and liver dissected out for the detection of fragmented DNA, DNA damage, and assay of oxidative stress markers. Moreover, expression levels of ß-Cat and p53 genes were measured by a real-time quantitative polymerase chain reaction. Results: The results showed a significant difference in MSG-treated group at the 2-time intervals than the control one regarding parameters of oxidative stress, and these were accompanied by a significant decline in glutathione (GSH) and a ratio of oxidized and reduced GSH in both tissues. Significant elevation of laddered DNA and oxidative DNA damage was observed in groups treated with MSG. In addition, a significant decline in gene expression of ß-Catenin in liver and brain tissues with elevations in the gene expression of p53 in the brain. Furthermore, the p53 gene in liver tissue was significantly upregulated in mice administered MSG for 15 days and was downregulated after 30 days of MSG intake compared with the control. Conclusion: According to our results, oral consumption of MSG leads to oxidative stress-mediated DNA damage and apoptosis.

CYTOGENETIC BIOMONITORING AND IMMUNOCYTOCHEMICAL STUDIES ON CAR SPRAY PAINTERS IN ENUGU METROPOLIS, ENUGU STATE, NIGERIA

Objective: This study assessed the cytogenetic damage associated with occupational exposure to paints by evaluating exfoliated buccal epithelial cells from car spray painters in Enugu metropolis using some biological markers. Methods: A total of 352 apparently healthy males, comprising 200 car spray painters and a control group of 152 individuals, participated in the study. Buccal smears were obtained from each participant and were stained using hematoxylin and eosin technique. A total of 1000 cells per individual were scored under light microscopy to determine the frequencies of micronuclei (MN) and binucleate cells (BNC). Structured questionnaires were used to obtain relevant participant information. Expression patterns of Ki-67and p53 genes on the buccal cells were determined by immunocytochemical methods. Results: Car spray painters had significantly increased frequencies of MN and BNC (*p<0.05) when compared to the control subjects. Paint sprayers aged over 35 years had higher buccal cell MN frequency when compared to those <25 years. Furthermore, car spray painters who have worked for ≥15 years had higher frequencies of MN when compared to those who had worked for <5 years (*p<0.05). Smoking and alcohol consumption increased the MN frequency of the car spray painters (*p<0.05). There was no expression of p53 and Ki-67 genes in the buccal cells of both control and exposed subjects. Conclusion: Car spray painters in Enugu metropolis may be occupationally exposed to substances capable of inducing genotoxic changes which manifested as increased frequency of MN in their buccal cells.