Sains Malaysiana 51(7)(2022): 2033-2046

http://doi.org/10.17576/jsm-2022-5107-08

Low Methylation of Matrix Metalloproteinase 1 (MMP1) is Associated with Preterm Labour in Malaysian Mothers

(Metilasi Rendah Matriks Metalloproteinase 1 (MMP1) dikaitkan dengan Kelahiran Bayi Pramatang dalam kalangan Ibu di Malaysia)

NURUL HAYATI MOHAMAD ZAINAL^1,*, NOR AZLIN MOHAMED ISMAIL² & NORFILZA M. MOKHTAR^3,*

¹Department of Human Anatomy, Faculty of Medicine and Health Sciences, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia

²Department of Obstetrics & Gynecology, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Center, Jalan Yaacob Latif, Bandar Tun Razak, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

³Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Preclinical Building, Jalan Yaacob Latif, Bandar Tun Razak, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

Received: 4 February 2021/Accepted: 7 January 2022

Abstract

Preterm births comprise 10.6% of livebirths worldwide and account for 35% of deaths among newborn babies. Understanding DNA methylation may offer basic knowledge in the understanding of pathogenesis of preterm labour. The study was undertaken to determine DNA methylation of matrix metalloproteinase 1 (MMP1) promoter in term and preterm labour using methylation-specific polymerase chain reaction (MSP). Thirty maternal venous blood samples (n=15 each) of term and preterm labour was subjected to bisulfite treatment prior to MSP. This result was then validated using DNA sequencing. Evaluation of the sequencing results by CpG islands analysis was performed using the ClustalW and SPSS software. Primers for MMP1 were located between -1226 and -1378 upstream from the transcription start site (TSS) that consisted five CpG islands. Preterm labour group had significantly lower methylated CpG islands with 39 out of total 75 (52%) compared to the term labour that has 49 out of 75 methylated CpG islands (65.33%) (t=0.694, p<0.05). Methylation occurred in 4 out of 5 methylated CpG islands in the MMP1 promoter where it only involved 2 preterm samples (13.33%) and 7 term samples (46.47%). This data suggested there were significant lower percentage of methylated MMP1 in preterm labour. Higher percentage of methylated MMP1 as observed in the term labour, will probably reduce the expression of MMP1, thus maintaining fibrillar collagen strength on the amniotic membrane and subsequently maintain the pregnancy till term. In conclusion, preterm labour has higher percentage of methylated CpG compared with term labour in MMP1 gene. 

Keywords: CpG islands; DNA methylation; matrix metalloproteinase 1; methylation-specific PCR; preterm labour

Abstrak

Kelahiran bayi pramatang merangkumi 10.6% kelahiran hidup di seluruh dunia dan menyumbang 35% kematian dalam kalangan bayi yang baru lahir. Pemahaman mengenai metilasi DNA boleh menyumbang kepada asas pengetahuan dalam memahami patogenesis kelahiran bayi pramatang. Kajian ini dilakukan untuk menentukan metilasi DNA matriks metalloproteinase 1 (MMP1) promoter dalam kelahiran bayi matang dan bayi pramatang menggunakan analisis bisulfit, metilasi khusus tindak balas rantaian polimerase (MSP). Tiga puluh sampel darah vena ibu (n=15 setiap kumpulan) daripada kelahiran bayi matang dan bayi pramatang menjalani rawatan bisulfit dan seterusnya MSP. Keputusan MSP disahkan menggunakan penjujukan DNA. Penilaian keputusan penjujukan bagi analisis gugusan CpG dibuat menggunakan perisian ClustalW dan SPSS. Primer bagi MMP1 terletak di antara -1226 dan -1378 daripada lokasi permulaan transkripsi (TSS) gen MMP1 yang mengandungi lima gugusan CpG. Kumpulan kelahiran bayi pramatang mempunyai gugusan CpG metilasi yang signikannya lebih rendah iaitu 39 daripada keseluruhan 75 (52%) berbanding kelahiran bayi matang yang mempunyai 49 daripada jumlah keseluruhan 75 CpG (65.33%) (t=0.694, p<0.05). Metilasi dikesan pada 4 daripada 5 gugusan CpG dalam promoter MMP1 dan ia hanya melibatkan 2 sampel pramatang (13.33%) dan 7 sampel kelahiran matang (46.47%). Data ini menunjukkan terdapatnya peratusan signifikan metilasi MMP1 yang lebih rendah dalam kelahiran bayi pramatang. Peratusan metilasi MMP1 yang lebih tinggi dalam kelahiran bayi matang berpotensi menyebabkan ekspresi MMP1 berkurangan dan mengekalkan kekuatan kolagen fibrilar membran amnion, seterusnya mengekalkan kehamilan sehingga tempoh matang. Kesimpulannya, kelahiran bayi pramatang mempunyai peratusan gugusan CpG metilasi yang lebih rendah berbanding kelahiran bayi matang bagi gen MMP1.

Kata kunci: Gugusan CpG; kelahiran bayi pramatang; matriks metalloproteinase 1; metilasi DNA; metilasi khusus tindak balas rantaian polimerase (MSP)

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^*Corresponding author; email: mz_nurul@upm.edu.my