Sains Malaysiana 49(1)(2020): 161-168

http://dx.doi.org/10.17576/jsm-2020-4901-20

DNA Methylation of Human Choline Kinase Alpha Gene

(Metilasi DNA Gen Alfa Kolina Kinase Manusia)

SITI AISYAH FATEN MOHAMED SA'DOM, NUR FARAH HANI AZEMI, MOHAMMAD SHAHRIZAL MOHAMMAD UMAR, YOKE HIANG YEE, WEI CUN SEE TOO & LING LING FEW*

School of Health Sciences, Health Campus, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan Darul Naim, Malaysia

Diserahkan: 12 Ogos 2018/Diterima: 17 Oktober 2019

Abstract

Increased level of choline kinase (CK) is a common feature in cancers and inhibition of this enzyme has been applied as anticancer strategy. DNA methylation of gene promoter especially at CpG island is associated with suppression of gene expression. Despite the importance of CK especially the alpha isoform in cancer pathogenesis, epigenetic regulation of cka expression has not been investigated. Hence, this study aimed to determine the effect of DNA methylation on cka promoter activity and gene expression by using hypomethylating (5-aza) and methylating (budesonide) agents. The level of DNA methylation in the second CpG island of cka promoter was determined by PCR-based method. 5-aza and budesonide increased the methylation of the selected CpG island compared to untreated control. Treatment with the drugs produced opposite effect, with 5-aza induced cka promoter activity and gene expression while budesonide suppressed the promoter activity and mRNA level of this gene. Deletion of a region containing the second CpG island on cka promoter resulted in significantly lower promoter activity. In conclusion, this study showed that DNA methylation could be one of the mechanisms that regulate the expression of cka gene.

Keywords: Choline kinase; DNA methylation; epigenetics; gene expression

Abstrak

Peningkatan tahap kolina kinase (CK) adalah ciri umum kanser dan perencatan enzim ini telah diguna sebagai strategi antikanser. Metilasi DNA promoter gen terutamanya pada pulau CpG telah dikaitkan dengan perencatan pengekspresan gen. Walaupun CK, terutamanya isoforma alfa, penting dalam patogenesis kanser, pengawalaturan epigenetik pengekspresan cka masih belum pernah dikaji. Maka, kajian ini bertujuan menentukan kesan metilasi DNA terhadap aktiviti promoter dan pengekspresan gen cka dengan menggunakan agen hipometilasi (5-aza) dan metilasi (budesonida). Tahap metilasi DNA pada pulau CpG kedua pada promoter cka telah ditentukan dengan kaedah berdasarkan PCR. 5-aza dan budesonida meningkatkan tahap metilasi pulau CpG yang dikaji berbanding kawalan tanpa rawatan. Rawatan dengan agen ini menghasilkan kesan berlawanan dengan 5-aza merangsang aktiviti promoter dan pengekspresan gen cka manakala budesonida merencat aktiviti promoter dan tahap mRNA gen ini. Pemadaman salah satu kawasan yang mengandungi pulau CpG kedua pada promoter cka menyebabkan penurunan secara signifikan aktiviti promoter. Kesimpulannya, kajian ini menunjukkan metilasi DNA mungkin merupakan salah satu mekanisme yang mengawalatur pengekspresan gen cka.

Kata kunci: Epigenetik; kolina kinase; metilasi DNA; pengekspresan gen

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*Pengarang untuk surat-menyurat; email: fewling@usm.my