PENAMBATAN MOLEKUL SENYAWA VOLATIL EKSTRAK DIKLOROMETANA SAMBILOTO TERHADAP JALUR PENSINYAL EPIDERMAL GROWTH FACTOR RECEPTOR: Penambatan Molekul Senyawa Volatil Ekstrak Diklorometana Sambiloto Terhadap Onkoprotein Human Epidermal Growth Factor Receptor

EGFR oncoprotein has been commonly studied in combating cell cancer development. However, until this moment the EGFR inhibitors were reported to cause other health issues. Plant extracts are trusted as the potential inhibitor of EGFR expression level without affect our health. We previously observed seventeen volatile compounds in the dichloromethane extract of Andrographis paniculata. Our objective was to analyze the interaction of volatile compounds from the dichloromethane extract of Andrographis paniculata on human EGFR pathway. In silico technique with ligand-receptor molecular docking was used in this study. The structure of volatile compounds was set as the ligands. EGFR, PIK3CA, KRAS-GTP, BRAF V600E, and AKT protein structures were assigned as the receptors. PyRx and Biovia Discovery Studio were used in this docking study. Drug-likeness and lead-likeness properties were appraised by SwissADME and Pre-ADME/Tox web tools. Beta-amyrin and stigmasterol showed the highest binding affinity to EGFR oncoproteins. PIK3CA, AKT, and KRAS-GTP, BRAF V600E was bound to beta-amyrin and stigmasterol, respectively. Those compounds structurally showed drug-likeness and non-mutagenic. Briefly, beta-amyrin and stigmasterol will be potentially used as the inhibitors of selected oncoproteins. In vitro technique and animal model are suggested to be performed to validate the mutagenic mechanisms of beta-amyrin and stigmastero

Novel Compound Heterozygous Mutations in the TRAPPC9 Gene in Two Siblings With Autism and Intellectual Disability

Autism spectrum disorder (ASD) is a highly heterogeneous neurodevelopmental disorder with many contributing risk genes and loci. To date, several intellectual disability (ID) susceptibility genes have frequently been identified in ASD. Here, whole exome sequencing was carried out on a proband with ASD and identified compound heterozygous mutations of the TRAPPC9, which plays a role in the neuronal NF-κB signaling pathway. These mutations consisted of a novel frameshift mutation (c.2415_2416insC, p.His806Profs∗9) and a rare splice site mutation (c.3349+1G>A) that were segregated from an unaffected father and unaffected mother, respectively. These two heterozygous mutations were also identified in the patient’s older brother with ID. Quantitative RT-PCR revealed a significant reduction of TRAPPC9 transcript in two siblings. This study first describes compound heterozygous mutations of the TRAPPC9 gene in two siblings with ASD and ID, which is notable as only homozygous mutations or compound heterozygous for copy number variations and rare variant in this gene have been reported to date and associated with autosomal recessive intellectual disability. The two siblings carrying compound heterozygous TRAPPC9 mutations presented with ID, developmental delay, microcephaly and brain abnormalities similarly to the clinical features found in almost cases with homozygous TRAPPC9 mutation in previous studies. Together this study provides evidence that clinical manifestations of TRAPPC9 mutations as seen in our patients with ID and autism may be broader than previous case reports have indicated

Vatica diospyroides Symington type LS Root Extract Induces Antiproliferation of KB, MCF-7 and NCI-H187 Cell Lines

Purpose: To investigate the therapeutic efficacy of V. diospyroides Symington type LS root extract as a chemopreventive agent against various cancer cell lines.Methods: Acetone root extract was evaluated for in vitro cytotoxicity against KB (oral cavity cancer), MCF-7 (breast cancer), and NCI-H187 (small cell lung cancer), using Resazurin microplate assay (REMA). Toxicity against a representative normal cells, Vero (African green monkey kidney), was assessed using green fluorescence protein (GFP)-based assay.Results: V. diospyroides root extract showed significant cytotoxic effects on KB and MCF-7 cell lines in a dose-dependent manner with IC50 of 35.05 ± 1.45 and 36.63 ± 3.40 μg/mL, respectively. NCI-H187 was not significantly inhibited (≤ 19.39 % inhibition) at the concentrations tested. IC50 against Vero cells was outside the concentration range of 0.2 - 50 μg/mL.Conclusion: These results indicate that the root extract of V. diospyroides has in vitro cytotoxic effect on human oral cavity cancer and breast cancer cells. No toxic effect on normal cells was observed. Thus, the extract may provide bioactive substances for human cancer therapy.Keywords: Breast cancer, Oral cavity cancer, Lung cancer, Cytotoxicity, Vero cells, Vatica diospyroide

Characterization of a Novel Binding Protein for Fortilin/TCTP — Component of a Defense Mechanism against Viral Infection in Penaeus monodon

The Fortilin (also known as TCTP) in Penaeus monodon (PmFortilin) and Fortilin Binding Protein 1 (FBP1) have recently been shown to interact and to offer protection against the widespread White Spot Syndrome Virus infection. However, the mechanism is yet unknown. We investigated this interaction in detail by a number of in silico and in vitro analyses, including prediction of a binding site between PmFortilin/FBP1 and docking simulations. The basis of the modeling analyses was well-conserved PmFortilin orthologs, containing a Ca2+-binding domain at residues 76–110 representing a section of the helical domain, the translationally controlled tumor protein signature 1 and 2 (TCTP_1, TCTP_2) at residues 45–55 and 123–145, respectively. We found the pairs Cys59 and Cys76 formed a disulfide bond in the C-terminus of FBP1, which is a common structural feature in many exported proteins and the “x–G–K–K” pattern of the amidation site at the end of the C-terminus. This coincided with our previous work, where we found the “x–P–P–x” patterns of an antiviral peptide also to be located in the C-terminus of FBP1. The combined bioinformatics and in vitro results indicate that FBP1 is a transmembrane protein and FBP1 interact with N-terminal region of PmFortilin

กลไกการทำงานของ Fortilin/TCTP ที่เกี่ยวข้องกับกระบวนการตายของเซลล์

Thesis (Ph.D., Biochemistry)--Prince of Songkla University, 200

Cytotoxic Activity of Piper cubeba Extract in Breast Cancer Cell Lines

This study aimed to evaluate the cytotoxicity of a crude extract of Piper cubeba against normal and breast cancer cell lines. To prepare the extract, P. cubeba seeds were ground, soaked in methanol and dichloromethane and isolated by column chromatography. Fractions were tested for cytotoxicity effects on normal fibroblast (L929), normal breast (MCF-12A) and breast cancer cell lines (MCF-7, MDA-MB-468 and MDA-MB-231). The most effective fraction was selected for DNA fragmentation assay to detect apoptotic activity. The results showed that the methanolic crude extract had a higher cytotoxic activity against MDA-MB-468 and MCF-7 than a dichloromethane crude extract. Then, the methanolic crude extract was separated into six fractions, designated A to F. Fraction C was highly active against breast cancer cell lines with an IC50 value less than 4 μg/mL. Therefore, Fraction C was further separated into seven fractions, CA to CG. The 1H-NMR profile showed that Fraction CE was long chain hydrocarbons. Moreover, Fraction CE demonstrated the highest activity against MCF-7 cells with an IC50 value of 2.69 ± 0.09 μg/mL and lower cytotoxicity against normal fibroblast L929 cells with an IC50 value of 4.17 ± 0.77 μg/mL. Finally, DNA fragmentation with a ladder pattern characteristic of apoptosis was observed in MCF-7, MDA-MB-468, MDA-MB-231 and L929 cells, but not in MCF-12A cells

Sermsum Graidist

Thesis (M.S.R.M.)--Prince of Songkla University, 200

Study of fortilin interacting proteins in a human skeletal muscle using the yeast two-hybrid system

Fortilin is implicated in development, cellular processes and malignant transformation. However, a unifying picture ofproteins in key processes pertaining to fortilin function is not yet emerging. To investigate the potential interactions, humanfortilin was expressed in the yeast cells, and used to screen for fortilin binding proteins in a human skeletal muscle. Yeastcells were transformed in the sequential procedure using yeast two-hybrid expression vector (pAS2-1) containing full-lengthhuman fortilin as ‘bait’ for the first transformation, and pGAD10 vector containing a muscle cDNA library for the secondtransformation. In addition, a direct interaction of fortilin with known proteins, Ca2+-ATPase, creatine kinase, glycogenphosphorylase, and troponin C was evaluated. -Galactosidase activity was assayed as an index of interaction betweenfortilin and the potential target proteins whereas yeast mating strategy was used to eliminate false positive and to reconfirmthe actual binding. From this analysis, eukaryotic translation elongation factor-1 delta (guanine nucleotide exchange protein)was identified as a putative fortilin interacting protein

การวิเคราะห์โปรติโอมิกส์ของ WT1 ที่ถูกยับยั้งด้วย siRNA ในเซลล์มะเร็งเต้านมเพาะเลี้ยงชนิด MCF-7 และ MDA-MB-468 : รายงานวิจัยฉบับสมบูรณ์

Prince of Songkla Unversit

Study of fortilin interacting protein in a skeletal muscle using the yeast two-hybrid system

Prince of Songkla Universit