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All Journal Dental Journal (Majalah Kedokteran Gigi) Bhakta Dental Journal
Puteri, Astari
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Biochemistry, Genetics & Molecular Biology Dentistry Immunology & microbiology Materials Science & Nanotechnology Medicine & Pharmacology

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Mdm2 Gene as Transcriptional Protein Antagonist of P53 in OSCC with Mir-218 Expression Hanif, Hilmy Irsyadi; Zafar, Anis Fatimah; Puteri, Astari; Wati, Sisca Meida; Rahayu, Retno Pudji
Bhakta Dental Journal Vol 2 No 02 (2024): Desember
Publisher : Fakultas Kedokteran Gigi IIK Bhakti Wiyata, Kediri.

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.56710/bhaktadent.v2i02.47

Abstract

Background: Murine double minute-2 (Mdm-2) gene expressed overly to the various type of human cancers. Murine double minute-2 associates with p53 affecting negatively with its gene products for activation of its transcriptional function. These genes mainly expressed in oral squamous cell carcinomas with p53 mutation condition. MicroRNAs nowadays becoming a promising diagnostic and prognostic biomarkers for malignancy through the expression and regulating the transcriptional function. It is reported that mir-218 expression may indicate the occurrence of OSCC. Purpose: The literature describes the putative pathway and function of mdm2 gene as transcriptional antagonist function of p53 in OSCC of mir-218 expression. Review(s): The relationship of mdm2 with mir-218 expression in OSCC may indicate the alternative mechanism of p53 dysfunction condition. Downregulated mir-218 will tend less binding to promoter of p53 causing the mutant state. The overexpression of mdm-2 may constitute inactivation of p53. Conclusion: The study findings may suggest mdm2 protein causing in OSCC due alternative mechanism of p53 as downregulated expression of mir-218.
Optimization of proteinase K incubation protocol duration during DNA extraction from oral squamous cell carcinoma FFPE samples Meizarini, Asti; Puteri, Astari; Yasan, Yanna Debby Restifanny; Hussaini, Haizal Mohd
Dental Journal (Majalah Kedokteran Gigi) Vol. 56 No. 4 (2023): December
Publisher : Faculty of Dental Medicine, Universitas Airlangga https://fkg.unair.ac.id/en

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/j.djmkg.v56.i4.p233-237

Abstract

Background: Formalin-fixed paraffin-embedded (FFPE) specimen archives are a valuable source of sample material for molecular biological analysis. However, the DNA isolated from FFPE samples is usually low in concentration and fragmented. Thus, it is necessary to optimize the FFPE DNA extraction protocol to obtain the best results. Proteinase K incubation is undoubtedly crucial in DNA extraction procedures, but this step is often not well explained in the manufacturer's manual. Purpose: This study aimed to find the optimal duration for proteinase K incubation protocols to achieve the highest DNA yields. Methods: Fifteen paraffin blocks of Oral Squamous Cell Carcinoma (OSCC) specimens were obtained, and the cancerous areas were microdissected into smaller cuts for DNA extraction. The samples were randomly divided into three groups (n=5) and subjected to three different proteinase K incubation protocols: one-hour incubation at 56ºC as per the manufacturer's instructions (Group I), 24-hour incubation at 56ºC (Group II), and 48 hours at room temperature with an additional four hours at 56ºC (Group III). The extracted DNA was then quantified using a Nanodrop spectrophotometer. The recorded data were analyzed using ANOVA-LSD. Results: The highest DNA concentration was found in Group III (107.74 ± 41.92), which was significantly higher compared to Group II (59.46 ± 30.32) and Group I (6.46 ± 1.97) (p<0.05). Conclusion: In conclusion, modifying the duration of proteinase K incubation protocols can lead to different DNA yield results. In this study, the most optimized protocol for proteinase K incubation, resulting in the highest DNA yields, was 48 hours at room temperature with an additional four hours at 56ºC.
Co-Authors Asti Meizarini Hanif, Hilmy Irsyadi Hussaini, Haizal Mohd Retno Pudji Rahayu Sisca Meida Wati Yasan, Yanna Debby Restifanny Zafar, Anis Fatimah