Deteksi mutasi gen TSHR pada penderita penyakit graves di Puskesmas Sukoharjo Kabupaten Sukoharjo

SLAMET, R. Agus Wibowo, Prof. drh. Widya Asmara, SU.,Ph.D

2010 | Tesis | S2 Ilmu Kedokteran Tropis

Hipertiroid terjadi karena kelenjar tiroid memproduksi hormon tiroid secara berlebihan sehingga menekan TSH. Hampir 60%-80% hipertiroid disebabkan oleh penyakit Graves. Prevalensi penyakit Graves pada perempuan lebih dari 2% dan kejadian penyakit ini pada wanita sepuluh kali dibanding pria. Penyakit Graves pada wanita terjadi pada usia 20-50 tahun. Beberapa gen telah diindikasikan sebagai penyebab penyakit Graves. TSHR terletak pada kromosom 14q31 dan mempunyai hubungan yang erat pada kejadian penyakit Graves. Tiga polimorfisme pada gen TSHR sudah berhasil diidentifikasi pada Asp36His, Pro52Thr, dan Asp727Glu. Codon D727E dilaporkan mempunyai hubungan dengan penyakit Graves. Tujuan dari penelitian ini adalah untuk mendeteksi mutasi pada Codon D727E gen TSHR pada penderita penyakit Graves. Enam belas responden terduga penyakit Graves diambil darahnya untuk diperiksa TSH, FreeT4, antibodi Tiroglobulin, antibodi Tiroperoxidase dan isolasi DNA. Gen TSHR codon D727E diamplifikasi dengan Polymerase Chain Reaction (PCR). Untuk mendeteksi mutasi hasil PCR dipotong dengan enzim Eco72I endonuklease restriksi (PCR-RFLP), PCR-Single Strand Conformation Polymorphism (PCR-SSCP), dan Sequencing. Hasil PCR-RFLP, PCR-SSCP dan Sequencing menunjukkan bahwa tidak ada mutasi pada gen TSHR codon D727E. Codon D727E tidak berhubungan dengan penyakit Graves pada populasi di Sukoharjo. Untuk mengetahui kemungkinan jenis mutasi lain pada gen TSHR yang berhubungan dengan penyakit ini perlu dilakukan penelitian lebih lanjut.

Hyperthytiroid is the rules of execessive thyroid function, with this happened of increased of thyroid hormone that suppressed TSH. Major etiologies of hyperthyroidism caused by Graves’disease. Graves’disease account for 60%-80% of hyperthyroidism. Graves’disease accour in up to 2% of women but is one-tenth as frequent in men. Graves’disease reraly accour between 20-50 year old. Graves’disease is polygenic thyroid syndrome. Some of the genes implicated in this pathogenesis may encode thyroid stimulating hormones receptors (TSHR). The TSHR gene is located on chromosome 14q31, an area in which a Graves’ disease susceptibility locus (GD) has been mapped. The GD locus is specifically linked to Graves’ disease. TSHR polymorphisms, resulting in amino acid substitutions, have been identified. Two of these are located in the extracellular domain of the receptor (Asp36His and Pro52Thr), and one is located in the intracellular domain (Asp727Glu). A germline mutation of TSHR codon 727 has been reported to be associated with Graves’disease. The aim of this research was analized the mutation of Codon D727E of TSHR gene at Graves' disease patient. Sixtenth respondens with suspect Graves' disease were analyzed. The blood were collected for TSH, FreeT4, Tyroglobulin Antibody and Tyroperoxidase Antibody analyzed and DNA isolation. The polymorphic region of the target genes(gene TSHR codon 727) were amplified by polymerase chain reaction (PCR) on the basic of genomic DNA isolated blood of 16 respondent from Sukoharjo public health center. To detect the D727E mutation, the PCR product were additionally digested with Eco72I restriction endonuklease (PCR-RFLP), Polymerase Chain Reaction-Single Strand Conformation Polymorphism (PCR-SSCP) and for final confirmation by PCR-Sequencing. The result of analyzing codon D727E by PCR-RLFP, PCR-SSCP and Sequencing method showed there is no mutation in gen TSHR codon D727E. The codon D727E variant of TSHR gene is not associated with Graves’disease in Sukoharjo population. The possibility that mutation may occur at another genes need to be further analyzed.

Kata Kunci : Hipertiroid,Penyakit graves,TSHR codon D727E,PCR,RFLP,PCR,SSCP, Hyperthyroidism, Graves’ disease, TSHR codon D727E, PCR-RFLP, PCR-SSCP, Sequencing