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Abstrak :
Latar Belakang: Endometriosis merupakan penyakit multifaktorial yang mempengaruhi 10% wanita usia subur. Diketahui bahwa gen EGFR dan MMP-2 mengalami peningkatan ekspresi pada endometriosis sehingga memiliki peran dalam perkembangan endometriosis, dan gen yang dapat meregulasi sitoskeleton. Tujuan dari penelitian ini adalah untuk mengevaluasi hubungan antara tingkat metilasi gen EGFR dan MMP-2 dengan ekspresi mRNA-nya pada jaringan endometriosis peritoneum. Metode Penelitian: Penelitian ini menggunakan desain cross sectional. Sampel yang digunakan sebanyak 20 wanita endometriosis dan 20 wanita bukan endometriosis yang usianya sekitar 20-45 tahun. Pada wanita endometriosis diambil jaringan endometriosis peritoneum dengan tindakan laparoskopi, sedangkan 20 wanita bukan endometriosis diambil jaringan endometrium normal dengan tindakan mikrokuretase. Tingkat metilasi DNA gen EGFR dan MMP-2 dianalisis dengan metode Methylation Specific PCR (MSP) dan Ekspresi mRNA gen EGFR dan MMP-2 dianalisis dengan metode qRT-PCR. Hasil: Tingkat metilasi DNA pada gen EGFR dan MMP-2 mengalami hipermetilasi. Pada gen EGFR, tingkat metilasi DNA antara jaringan endometriosis peritoneum dibandingkan dengan jaringan endometrium normal terdapat perbedaan yang bermakna (p=0,001), sedangkan pada gen MMP-2 tingkat metilasi DNA-nya tidak terdapat perbedaan yang bermakna (p=0,596) antara jaringan endometriosis peritoneum dibandingkan dengan jaringan endometrium normal. Nilai ekspresi relatif mRNA EGFR dan MMP-2 mengalami peningkatan ekspresi pada jaringan endometriosis peritoneum. Penelitian ini tidak menunjukkan korelasi yang bermakna antara tingkat metilasi dengan tingginya ekspresi mRNA baik gen EGFR maupun MMP-2. (gen EGFR (p=0,947 dan r=-0,016) dan gen MMP-2 (p=0.769 dan r=0.070) Kesimpulan: Tingginya ekspresi mRNA EGFR dan gen MMP-2, kemungkinan bukan hanya disebabkan karena faktor metilasi DNA, melainkan faktor lainnya.

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Background: Endometriosis is a multifactorial disease that affects 10% of women of childbearing age. It is known that the EGFR and MMP-2 genes have increased expression in endometriosis and thus have a role in the development of endometriosis, and genes that can regulate the cytoskeleton. The purpose of this study was to evaluate the relationship between the level of methylation of the EGFR and MMP-2 genes with their mRNA expression in peritoneal endometriosis tissue. Method: The study used a cross sectional design. The sample used was 20 women with endometriosis and 20 women without endometriosis who were around 20-45 years old. In endometriosis women are taken to peritoneal endometriosis tissue by laparoscopic, while 20 women without endometriosis are taken to normal endometrial tissue by microcuretase. The levels of EGFR and MMP-2 gene methylation were analyzed by the Methylation Specific PCR (MSP) method and the mRNA expression of the EGFR and MMP-2 genes were analyzed by the qRT-PCR method. Results: The level of DNA methylation in the EGFR and MMP-2 genes was hypermethylated. In the EGFR gene between peritoneal endometriosis tissue compared to normal endometrial tissue there were significant differences (p=0,001), whereas in the MMP-2 gene there was no significant difference (p=0.596) between peritoneal endometriosis tissue compared to normal endometrial tissue. The relative expression value of EGFR and MMP-2 mRNA has increased expression in peritoneal endometriosis tissue. This study did not show a significant correlation between the level of methylation and the high mRNA expression in both the EGFR and MMP-2 genes. (EGFR gene (p=0.947 and r=-0.016) and MMP-2 gene (p=0.769 and r=0.070) Conclusion: The high expression of EGFR mRNA and MMP-2 gene, the possibility is not only due to hypermethylation factors, but other factors.

Abstrak :
Latar belakang: Telah dilaporkan bahwa terdapat perubahan pada ekspresi dari ribuan gen di jaringan endometrium endometriosis, termasuk diantaranya adalah gen FN1 dan RAC1. Perubahan ekspresi gen tersebut dapat disebabkan oleh mekanisme epigenetik seperti perubahan tingkat metilasi DNA pada gen. Tujuan: Mengetahui tingkat metilasi DNA pada gen FN1 dan RAC1 serta ekspresi mRNAnya pada jaringan endometrium subjek endometriosis dan nir-endometriosis. Metode: Penelitian ini merupakan cross sectional dengan jumlah sampel sebanyak 40 dari jaringan endometrium subjek endometriosis dan subjek nir-endometriosis. Sampel diambil dengan teknik mikrokuretase di RSUPN Ciptomangunkusumo dan RS Fatmawati Jakarta. Pada jaringan kemudian dilakukan isolasi DNA dan RNA. Pada isolat DNA dilakukan konversi bisulfit, MSP, elektroforesis dan analisis intensitas pita menggunakan software ImageJ untuk mendapatkan data persentase tingkat metilasi DNA. Pada isolat RNA dilakukan qRT-PCR untuk mendapatkan ekspresi relatif mRNA gen FN1 dan RAC1. Hasil: Analisis persentase tingkat metilasi DNA promotor menunjukkan terdapat perbedaan bermakna (p=0,022) pada gen FN1 pada pasien endometriosis (37,95%) dibandingkan nir-endometriosis (59,22 %), sedangkan pada gen RAC1 tidak terdapat perbedaan bermakna (p=0,63) dengan tingkat metilasi subjek endometriosis (28.45%) dan subjek nir-endometriosis (26.11%). Penelitian ini juga melaporkan terjadinya peningkatan ekspresi relatif mRNA gen FN1 dan RAC1 dibandingkan dengan subjek nir-endometriosis, namun secara statistik tidak terdapat perbedaan bermakna (p>0,05). Tidak terdapat korelasi bermakna antara tingkat metilasi gen FN1 dan RAC1 dengan ekspresi mRNAnya. Kesimpulan: Terjadi penurunan tingkat metilasi yang bermakna pada gen FN1 di jaringan endometrium endometriosis, namun tidak berkorelasi dengan peningkatan mRNA nya. Tidak terdapat perbedaan bermakna tingkat metilasi dan ekspresi mRNA pada gen RAC1 di jaringan endometrium subjek endometriosis dibandingkan dengan nir endometriosis.

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It has been reported that there was a changes in the expression of thousands of genes in endometrial endometriosis tissues, including the FN1 and RAC1 genes. Changes in gene expression can be caused by epigenetic mechanisms such as DNA methylation in genes. Objective: To determine the level of DNA methylation in FN1 and RAC1 genes and their mRNA expression in endometrial tissue of endometriosis and non-ndometriosis. Method: This study was designed as cross sectional with a total sample of 40 of endometrial tissues in the subject of endometriosis and non-endometriosis. Samples were taken by microcuretase at Ciptomangunkusumo and Fatmawati Hospital, Jakarta. DNA and RNA was isolated. DNA isolates were converted by bisulfite procedure, MSP conversion, electrophoresis, analyzed intensity of the band which appeared on gel electrophoresis using ImageJ software to obtain the percentage data of DNA methylation level. In RNA isolates, it was analyzed using qRT-PCR methode to obtain the relative mRNA expression level. Results: Analysis of percentage of DNA methylation level showed significant differences (p=0.022) in the FN1 gene (37.95%) compared to non-endometriosis (59.22%), whereas in the RAC1 gene there was no significant difference (p=0,63) with methylation level of endometriosis subjects (28.45%) and non-endometriosis subjects (26.11%). For relative mRNA expression of FN1 and RAC1 genes showed no significant differences (p> 0.05). For correlation in endometrial endometriosis showed no significant between the rate of methylation of the FN1 and RAC1 genes with their mRNA expression. Conclusion: There was a significant decrease in DNA methylation level of FN1 gene in endometrial endometriosis tissues, but it did not correlate with the increasing in its mRNA expression. There was no significant difference in DNA methylation level and mRNA expression of RAC1 gene in endometrial tissues of endometriosis subjects compared to non-endometriosis.

Abstrak :
ABSTRAK
Latar belakang: Salah satu faktor yang dicurigai berperan dalam mekanisme resisensi klopidogrel adalah faktor epigenetik seperti metilasi DNA. Individu dengan resistensi klopidogrel ini memiliki kecenderungan untuk mengalami luaran kardiovaskular yang lebih buruk. Nilai TIMI flow pasca IKPP telah diketahui berkaitan dengan luaran klinis pada pasien IMA-EST. Sampai saat ini belum ada penelitian yang menghubungan antara metilasi gen P2Y12 dengan penghambatan fungsi platelet dan nilai TIMI flow pasca IKPP pada pasien IMA EST. Tujuan: Untuk mengetahui hubungan antara metilasi gen reseptor P2Y12 terhadap fungsi penghambatan platelet dan nilai TIMI flow pasca IKPP pada pasien IMA EST. Metode: Sebanyak 118 pasien IMA-EST yang menjalani IKPP dan mendapatkan terapi klopidogrel dimasukkan kedalam populasi penelitian. Dilakukan pemeriksaan VerifyNow P2Y12 dan pemeriksaan metilasi P2Y12. Selanjutnya dilakukan analisis hubungan antara metilasi P2Y12 dengan nilai Verifynow P2Y12 dan TIMI flow pasca IKPP. Hasil: Dari seluruh subyek, 22% diantaranya termasuk klopidogrel nonresponder dan 30% memiliki nilai TIMI flow kurang dari 3. Terdapat 48% subyek yang tidak mengalami metilasi dan 19% subyek mengalami metilasi sempurna pada gen P2Y12. Tidak terdapat hubungan bermakna antara metilasi P2Y12 dengan nilai Verifynow P2Y12 dan TIMI flow pasca IKPP. Nilai Verifynow P2Y12 yang tinggi berhubungan dengan TIMI flow kurang dari 3 pasca IKPP (p=0,043). Kesimpulan: Tidak terdapat hubungan bermakna antara pola metilasi gen P2Y12 dengan penghambatan fungsi platelet dan nilai TIMI flow pasca IKPP. Pasien yangnon-responder terhadap klopidogrel berisiko untuk mendapatkan reperfusi miokard yang suboptimal.

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ABSTRACT
Background: Mechanism of clopidogrel resistance is not well understood yet. In the other hand, epigenetic modifications such as DNA methylation, are suspected to play role in clopidogrel resistence. Subject with high on treatment clopidogrel reactivity show worsen cardiovascular outcome. Meanwhile, TIMI flow after reperfusion are known to be related with poor outcome. Study that evaluate the relationship between methylation of P2Y12 gene with Platelet Reactivity and TIMI-flow after Primary Percutaneous Coronary Intervention (PPCI) in Patients With Acute ST-segment Elevation Myocardial Infarction in South East Asia Population has never been done. Objectives: to define whether methylation of P2Y12 gene and platelet reactivity may affect the myocardial perfusion after PPCI. Methods: There were 118 of STEMI patients who underwent PPCI and had received clopidogrel were recruited for the study. We measured platelet reactivity using Verifynow P2Y12 and Methylation of P2Y12 gene. The relationship among variables are assessed using statistic method. Results: Among 118 subject, 22% are clopidogrel nonresponder and 30% had TIMI flow less than 3. Median of Methylation degree was 15% with 48% subject were unmethylated, 19% subject had 100% methylation. There are no relationship between methylation of P2Y12 gene with platelet reactivity and TIMI flow after PPCI among subjects. The value of Verifynow P2Y12 more than 208 were related TIMI flow less than 3 after PPCI (p=0,043). Conclusion: There are no relationship between methylation of P2Y12 gene with platelet reactivity and TIMI flow after PPCI among subjects. Clopidogel nonresponder subjects were more likely to have suboptimal reperfusion after PPCI

Abstrak :
Endometriosis sering dikaitkan dengan nyeri menstruasi dan nyeri pelvis. Penyakit ini terjadi sekitar 10-15% pada perempuan usia reproduksi . NGF, TRPA1 dan reseptor P2RX3 aktivitas gen terlibat dalam respon nyeri. Penelitian ini bertujuan untuk menganalisis tingkat ekspresi mRNA gen NGF, reseptor TRPA1 dan reseptor P2RX3 yang diduga disebabkan oleh perubahan tingkat metilasi DNA promoter gen tersebut, serta hubungannya pada intensitas nyeri subjek endometriosis.Sampel jaringan endometrium dan susukan endometriosis peritoneum diperoleh dari 20 subjek endometriosis, sementara jaringan endometrium kontrol diperoleh dari 20 subjek nir endometriosis. Metode yang digunakan untuk analisis metilasi DNA yaitu metode MSP dan perangkat lunak Image-J digunakan untuk menganalisis intensitas pita metilasi dari gen NGF, reseptor TRPA1 dan reseptor P2RX3R, selanjutnya digunakan metode qRT-PCR untuk analisis tingkat mRNA gen-gen tersebut. Penilaian intensitas nyeri dilakukan dengan menggunakan kuesioner standar skala penilaian numerik (NRS) melalui wawancara dengan pasien. Pada penelitian ini didapatkan hasil terdapat hubungan yang bermakna antara intensitas nyeri dan kejadian endometriosis (p>0,001). Hasil tersebut dibuktikan dengan terdapat perbedaan yang bermakna tingkat ekspresi relatif mRNA gen NGF, reseptor P2RX3 antara jaringan endometrium endometriosis dibandingkan dengan subjek nir endometriosis masing-masing nilai p= <0,05. Terdapat juga perbedaan yang bermakna tingkat metilasi DNA promotor gen NGF dan reseptor P2RX3 antara jaringan endometrium endometriosis dengan endometrium nir endometriosis (p<0,05). Selanjutnya, terdapat hubungan yang bermakna antara ekspesi mRNA NGF dan reseptor Reseptor P2RX3 dengan intensitas nyeri pada endometriosis (p<0,05), namun tidak terdapat hubungan antara tingkat metilasi DNA dengan ekspresi relatif mRNA gen NGF, reseptor reseptor TRPA1 pada endometriosis begitupun antara metilasi DNA dengan intensitas nyeri pada ketiga gen tersebut (p>0,05). Terjadi perubahan ekspresi relatif mRNA gen NGF, reseptor TRPA1 dan reseptor P2RX3 pada subjek endometriosis yang berhubungan dengan peningkatan intensitas nyeri pada endometriosis, namun mekanisme epigenetik metilasi DNA pada penelitian tersebut tidak berhubungan pada intensitas nyeri endometriosis. ...... Endometriosis is often associated with both cyclic menstrual pain and pelvic pain. It affects 10% of reproductive age women. NGF, TRPA1 and P2RX3 receptors gene activity are found to be involved in pain response. This study aims to analyze the methylation level of NGF gene, TRPA1 and P2RX3 receptors that might alter the mRNA expression in peritoneal endometriosis and endometrial tissue, as well as its correlation to the pain level in endometriosis patients.20 endometrial tissues and 20 peritoneal endometriosis tissues were obtained from patients, while 20 endometrium tissues as control were obtained from healthy women. First, each participant was given informed consent before the research begin. We used methylation specific PCR (MSP) and Image-J software to analyze the methylation level of NGF gene, TRPA1 and P2RX3 receptors; electrophoresis to analyze the band intensity; qRT-PCR to evaluate the mRNA level in each gene. Finally, we evaluated the pain level using the standardized questionnaire of numeric rating scale (NRS) by doing interviews with patients. In this study, it was found that there is a significant relationship between pain intensity and the incidence of endometriosis (p> 0.001). This results is proven by a significant difference in the mRNA expression level of NGF gene and P2RX3 receptor between endometrial endometriosis and endometrial non-endometriosis tissues, with each p value = <0.05. There is also a significant difference in the DNA methylation level of NGF gene and P2RX3 receptor between endometrial endometriosis and endometrial non-endometriosis tissues (p <0.05). There is a significant relationship between the mRNA expression level of NGF gene, P2RX3 receptor and pain intensity in endometrial endometriosis tissues (p <0.05). However, the results showed that there is no correlation between the DNA methylation level and the mRNA expression of NGF gene, TRPA1 and P2RX3 receptors. There is also no correlation between the DNA methylation level of NGF gene, TRPA1 and P2RX3 receptors and pain intensity in endometriosis tissue. There is an alteration of mRNA expression of NGF gene, TRPA1 and P2RX3 receptors which correlates to pain intensity in endometriosis patients. However, there is no correlation between DNA methylation level and pain intensity in endometriosis patients.

Abstrak :
Latar belakang: Resistensi klopidogrel merupakan salah satu faktor risiko penting terjadinya kejadian iskemik berulang pada pasien yang telah mendapat terapi anti platelet yang optimal. Penelitian ini bertujuan untuk mengetahui peran metilasi DNA gen CYP2C19 sebagai salah satu faktor epigenetik terhadap kejadian resistensi klopidogrel pada pasien infark miokard akut dengan elevasi segmen ST (IMA-EST) yang menjalani intervensi koroner perkutan primer (IKPP). Metode: Pasien IMA-EST yang menjalani IKPP diberikan antiplatelet klopidogrel dan menjalani pemeriksaan fungsi platelet degan VerifyNowTM. Kriteria untuk mendefinisikan resistensi klopidogrel adalah nilai PRU >208. Pemeriksaan metilasi DNA gen CYP2C19 dilakukan dengan metode bisulfite genomic sequencing technology. Data klinis, laboratorium, dan angiografik termasuk TIMI flow dikumpulkan untuk dilakukan analisis. Hasil: Dari 122 subjek, resistensi klopidogrel ditemukan pada 22% subjek. Kelompok dengan presentase metilasi DNA <50% mempunyai risiko lebih tinggi terjadinya resistensi klopidogrel (OR 4.5 IK95% 2.1-9.3, nilai p 0.018). Grup ini juga diketahui mempunyai TIMI flow post-IKPP yang suboptimal (OR 3.4 IK95% 1.3 - 8.7, nilai p 0.045). Kesimpulan: Hipometilasi DNA gen CYP2C19 meningkatkan risiko terjadinya resistensi klopidogrel dan luaran klinis yang lebih buruk. ......Background: Clopidogrel resistance is an important risk factor of recurrence of ischemic event after optimal antiplatelet therapy. We aimed to investigate the role of DNA methylation of CYP2C19 gene as one of epigenetic factor to the risk of clopidogrel resistance in ST-segment elevation myocardial infarction (STEMI) patients undergoing primary percutaneous coronary intervention (PPCI). Methods: STEMI patients undergoing PPCI were pretreated with clopidogrel, and their platelet function was measured using VerifyNowTM assay. The criteria for high on- treatment platelet reactivity (HPR) was defined according to the expert consensus criteria (PRU >208). DNA methylation of the CYP2C19 gene was performed using bisulfite genomic sequencing technology. Furthermore, clinical, laboratory and angiographic data including TIMI flow were collected. Result: Among 122 patients, clopidogrel resistance was found in 22% patients. After dividing into two groups, methylation <50% was associated with increased risk of clopidogrel resistance (OR 4.5 95%CI 2.1-9.3, P value 0.018). This group also found to have suboptimal post-PCI TIMI flow (OR 3.4 95%CI 1.3 - 8.7, P value 0.045). Conclusions: The lower DNA methylation level of the CYP2C19 gene increases the risk of clopidogrel resistance and subsequent poorer clinical outcome.

Abstrak :
ABSTRAK
Latar belakang: SOPK adalah gangguan endokrin yang hingga saat ini etiologinya masih belum jelas. Faktor epigenetik metilasi DNA, akhir-akhir ini mendapatkan perhatian dalam patogenesis SOPK. Gen HSD17B1 disebut sebagai "estrogenik" 17β-HSD karena mengkatalisasi langkah terakhir dalam biosintesis estrogen dengan secara istimewa mengurangi estrone, estrogen yang lemah untuk menghasilkan estrogen 17β-estradiol yang kuat. Kami berspekulasi cacat pada metilasi DNA mendorong deregulasi gen sehingga terjadi penurunan ekspresi mRNA HSD17B1, akhirnya menghasilkan estradiol yang tidak cukup pada pasien SOPK. Metode: Kami mengumpulkan total 60 pasien wanita. MSP untuk analisis metilasi DNA, qPCR untuk analisis ekspresi mRNA. Tujuan: Untuk menganlisis metilasi DNA pada kelompok pasien SOPKdan kelompok wanita sehat, ekspresi mRNA pada kelompok pasien SOPK dan kelompok wanita sehat, tingkat estradiol pada pasien SOPK dan kelompok wanita sehat, korelasi antara metilasi DNA dan ekspresi mRNA pada pasien SOPK, korelasi ekspresi mRNA pada pasien SOPKdan kadar serum estradiol. Hasil: Metilasi gen HSD17B1 pada wanita SOPK adalah 42,64% dan kelompok yang sehat menunjukkan 53,80%, p = 0,160 tidak signifikansi antara kedua kelompok. Nilai ekspresi relatif gen HSD17B1 adalah 0,70 kali lebih rendah dibandingkan dengan kelompok wanita sehat, p = 0,003 signifikansi antara kedua kelompok. Estradiol rata-rata pada kelompok SOPK25,78 pg / ml dan kelompok wanita sehat adalah 36,74 pg / ml. Korelasi tingkat metilasi DNA versus ekspresi mRNA pada pasien SOPK, tidak signifikan p = 0,076. Korelasi antara ekspresi mRNA gen HSD17B1 dan kadar serum estradiol, signifikansi p = 0,020. ;Semakin terjadi penurunan ekspresi mRNA, semakin rendah kadar serum estradiol.

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ABSTRACT
Background: PCOS is the most common endocrine disorder but its etiology remains unclear. Lately, epigenetic factors have gained considerable attention in the pathogenesis of PCOS, DNA methylation.  HSD17B1 is referred to as the "estrogenic" 17β-HSD because it catalyzes the final step in estrogen biosynthesis by preferentially reducing the weak estrogen estrone to yield the potent estrogen 17β-estradiol. We speculated defects in DNA methylation promote the deregulation of genes make decrease mRNA expression HSD17B1, finally produces not enough estradiol in PCOS patients. Methods: We collected a total of 60 female patients. MSP for DNA methylation analysis, qPCR for mRNA expression analysis. Aims: To investigate, DNA methylation in PCOS patients group and healthy women group, mRNA expression in PCOS patients group and healthy women group, estradiol level in PCOS patients and healthy women group, the correlation between DNA methylation and mRNA expression in PCOS patients, correlation mRNA expression in PCOS patients and estradiol serum level. Results: Methylated of HSD17B1 gene in PCOS women was 42.64 % and a healthy group showed 53.80 %, p=0.160 not significances between the two groups.  The relative expression value of the HSD17B1 gene was 0.70 fold lower compare with a healthy women group, p=0.003 significance between the two groups. The average estradiol in the PCOS group 25.78 pg/ml and the healthy women group is 36.74 pg/ml. Correlation of DNA methylation level versus mRNA expression in PCOS patients, not significance p=0.076. Correlation between mRNA expression of the HSD17B1 gene and estradiol serum level, significance p=0.020. (More decrease mRNA expression, more lower estradiol serum level).

Abstrak :
Latar belakang: Enzim methylenetetrahydrofolate reductase (MTHFR) terlibat dalam metabolism asam folat dan tipe allele mempengaruhi aktivitas enzim. Memberikan suplementasi asam folat kepada ibu hamil dapat mempengaruhi perubahan dalam derajat metilasi gen tertentu yang mempengaruhi kesehatan janin. Walaupun sudah banyak penelitian yang mempelajari peran asam folat sebagai donor dalam mekanisme epigenetik, namun penelitian pengaruh suplementasi besi-asam folat pada luaran kehamilan melalui pendekatan interaksi zat gizi-gen dalam desain penelitian longitudinal masih jarang. Penelitian ini bertujuan untuk mengetahui hubungan antara kadar serum asam folat pada ibu dan anak, dan derajat metilasi pada gen pencetak insulin-like growth factor (IGF2) yang dikenal terlibat dalam tumbuh kembang anak dan dapat digunakan sebagai penanda kemunculan penyakit Metode: Di tahun 2018, penelitian longitudinal dilakukan dengan mengunjungi 127 subyek termasuk anak yang dilahirkan dan mengikutsertakannya dalam penelitian. Enam puluh tujuh serum asam folat ibu selama hamil dan pasca melahirkan diperiksa, sementara serum asam folat anak dikumpulkan sebanyak 44 spesimen untuk pemeriksaan penanda darah. Pemeriksaan serum asam folat dengan menggunakan the liquid chromatography-mass spectrometry/mass spectrometry. Untuk pemeriksaan biomolekuler, tipe allele enzim MTHFR 677C>T and 1298A>C menggunakan Taqman polymerase chain reaction. Sementara metode pyrosequencing digunakan untuk menghitung DNA metilasi pada IGF2 pada anak. Hubungan antar variabel dianalisis menggunakan analisis regresi linier multivariat. Hasil:Tidak ada hubungan yang signifikan antara asupan asam folat dan serum asam folat ibu selama hamil, tiga tahun pasca melahirkan dan anak yang dilahirkan (p>0.05). Penelitian ini tidak dapat menunjukkan hubungan antara tipe allel dari MTHFR 677 C>T dan 1298 A>C dan serum asam folat (p>0.05). Serum asam folat selama hamil juga mempengaruhi status serum asam folat tiga tahun pasca melahirkan (p<0.05) dan status serum asam folat anak (p<0.05). Namun penelitian ini tidak dapat menunjukkan pengaruh status serum asam folat anak dengan DNA metilasi IGF2 pada anak (p>0.05). Simpulan: Serum asam folat selama hamil berkontribusi pada serum asam folat tiga tahun pasca melahirkan dan anak. Genotipe dari MTHFR gene at 677C>T and 1298 A>C kemungkinan tidak terlibat dalam metabolism asam folat pada ibu. Serum asam folat selama kehamilan tidak memiliki dampak pada status metilasi dari IGF2 pada wilayah differentially methylated region (DMR) untuk subyek anak. Namun, beberapa hal harus menjadi perhatian karena, secara statistik, jumlah subyek penelitian tidak memadai. Saran: Perlu dilakukan penelitian lanjutan yang melibatkan subyek lebih banyak dan metode yang lebih canggih dalam menentukan MTHFR dan metilasi DNA. ...... Background: Methylenetetrahydrofolate reductase, (MTHFR) enzyme is involved in folic acid metabolism, and their allele types affected its activity. Providing folic acid supplementation to pregnant mothers may influence the change in methylation level in specific genes that affect the susceptibility of disease of their offspring. Although folic acid's role as a donor in the epigenetic mechanism has been investigated, a longitudinal study exploring the influence of iron-folic acid supplementation on maternal dan birth outcome by the nutrient-gene interaction approach is lacking. Therefore, we investigated the relationship of serum folic acid level among the mothers and the children, and the imprinted insulin-like growth factor 2 (IGF2) methylation level that is known actively involved in growth and development in children and possibly utilized as a surrogate marker for the disease Methods: In 2018, the follow-up study conducted by re-visited 67 subjects and put the mother and their children included in the study. For each group, sixty-seven serums were collected for folic acid measurement for mothers during gestation and three-year post-partum. Furthermore, forty-four serums for children were gathered for biomarker measurement. Serum folics were measured by using liquid chromatography-mass spectrometry/mass spectrometry. Determining the genotype of the MTHFR enzyme in position 677C>T and 1298 A>C was used Taqman Polymerase Chain Reaction (PCR) method. The pyrosequencing method was utilized to quantify the methylation level of the IGF-2 of the children. The relationship analysis between variables using multivariate linear regression. Results: There was no relationship between the folic acid intake during gestation and serum folic acid of the mothers during pregnancy, three-year post-partum, and the children (p>0.05). There was no relationship between the allele type of MTHFR 677C>T and 1298A>C and serum folic acid status of the mother (p>0.05). The serum folic acid during pregnancy had a significant relationship to the serum folic acid three-year post-partum (p<0.05) as well as the serum folic acid of the children (p<0.05). There was no significant relationship between the serum folic acid of the children, serum homocysteine, and the methylation status of IGF2 of the children (p>0.05). Conclusion: The serum folic acid during pregnancy contributed to the serum folic acid three-year post-partum of mother and the children. The genotype of the MTHFR gene at 677C>T and 1298 A>C was possibly not involved in folic acid metabolism in the mother. Serum folic acid during pregnancy could not have an effect on the methylation status of the IGF2 in the differentially methylated region (DMR) area of the children. However, this conclusion needs to be taken in caution due to lack of study power Recommendation: Further cohorts studies with a large sample size and more advanced methods in determining the MTHFR enzyme and DNA methylation. Keyword: serum folic acid, genotyping MTHFR 677 C>T, MTHFR 1298 A>C, DNA methylation, IGF2.

Abstrak :
Metilasi DNA merupakan salah satu penyebab umum inaktivasi Mismatch Repair Gene (MMR). Gen MMR memperbaiki kesalahan penyisipan/penghapusan basa nukleotida pada proses sintesis DNA. Metilasi pada promoter gen MMR memiliki asosiasi dengan pembentukan kanker kolon, sehingga metilasi tersebut perlu diidentifikasi. Identifikasi gen MMR dapat dilakukan menggunakan teknik methylation-specific multiplex ligation-dependent probe amplification amplification (MS-MLPA). Prinsip dari teknik MS-MLPA yaitu amplifikasi probe yang menempel pada sekuens termetilasi. Tujuan dari penelitian ini yaitu untuk mengoptimasi teknik MS-MLPA dan mengidentifikasi metilasi gen MMR pada kanker kolon dengan teknik MS-MLPA. Penelitian ini menggunakan 27 sampel jaringan frozen kanker kolon yang telah tersedia di Biobank Rumah Sakit Kanker Dharmais (RSKD). Sampel tersebut dianalisis menggunakan probemix Mismatch Repair Gene [ME011-C1][C1-0518] yang telah didesain khusus untuk mendeteksi pada beberapa gen MMR yakni MLH1, PMS2, MSH6, dan MSH2. Hasil penelitian menunjukkan optimasi teknik MS-MLPA telah berhasil dilakukan, sehingga identifikasi metilasi pada gen MMR telah berhasil diperoleh pada 4 sampel pasien. Gen MMR tersebut yakni MLH1 dan MSH6, dengan persentase masing-masing 75% dan 25%. ......DNA methylation is one of the most common causes of mismatch repair gene (MMR) inactivation. The MMR gene corrects errors in the insertion/deletion of nucleotide bases in the DNA synthesis process. MMR gene promoter methylation has an association with the formation of colon cancer, so the methylation needs to be identified. Identification of the MMR gene can be done using the methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) technique. The principle of the MS-MLPA technique is the amplification of the probe attached to the methylated sequence. The purpose of this study was to optimize the MS-MLPA technique and identify MMR gene methylation in colon cancer using the MS-MLPA technique. This study used 27 samples of frozen colon cancer tissue that were available at the Dharmais Cancer Hospital Biobank (RSKD). The samples were analyzed using the Mismatch Repair Gene probemix [ME011-C1][C1-0518] which has been specially designed to detect several MMR genes, namely MLH1, PMS2, MSH6, and MSH2. The results show that the optimization of the MS-MLPA technique has been successfully carried out, so that the identification of methylation in the MMR gene has been successfully obtained in 4 patient samples. The MMR genes are MLH1 and MSH6, with a percentage of 75% and 25%, respectively. analyzed using probemix Mismatch Repair Gene [ME011-C1][C1-0518] which has been specifically designed to detect several MMR genes namely MLH1, PMS2, MSH6, and MSH2. The results showed that the optimization of the MS-MLPA technique was successful, so that identification of the methylation in the MMR gene was successfully obtained in 4 patient samples. The MMR genes are MLH1 and MSH6, with percentages of 75% and 25% respectively.