Pengaruh perlakuan cyclosporine A terhadap ekspresi gen CDR1, ERG2 dan RTA3 pada Candida albicans strain sensitif dan resisten fluconazole

KARTASASMITA, Ginanjar, dr. Tri Wibawa, Ph.D

2010 | Tesis | S2 Bioteknologi

Penelitian ini bertujuan untuk membandingkan ekspresi gen CDR1, ERG2 dan RTA3 pada Candida albicans strain sensitif dan resisten fluconazole isolat klinis Indonesia, untuk mengetahui pengaruh Cyclosporine A (CY A) terhadap ekspresi tiga gen tersebut baik pada strain sensitif dan resisten, serta melihat hubungan kekerabatan (filogenetik) antara dua strain tersebut berdasarkan sekuen gen 18S rRNA. Metode yang digunakan dalam menganalisis ekspresi gen adalah kombinasi RT-PCR dengan densitometri sedangkan untuk melihat hubungan kekerabatan digunakan metode analisis sekuen gen 18S rRNA menggunakan bantuan program Bioedit dan MEGA 4.0.2. Hasil analisis ekspresi gen menunjukan bahwa secara alami semua sampel C. albicans strain resisten tidak dapat mengekspresikan gen CDR1, ERG2 dan RTA3, sedangkan pada strain sensitif pada umumnya dapat mengekspresikan ketiga gen tersebut. Perlakuan CY A menyebabkan strain resisten R781 dan strain sensitif S300, S662, S966, S1023 dan S1034 mampu meningkatkan semua ekspresi gen yang dikaji. Hasil analisis filogenetik berdasarkan sekuen gen 18S rRNA menunjukan semua strain yang dikaji memiliki persamaan sekuen gen. Dari penelitian ini dapat disimpulkan bahwa secara alami ekspresi gen CDR1, ERG2 dan RTA3 antara Candida albicans strain resisten dan sensitif fluconazole adalah berbeda, pengaruh CY A adalah meningkatkan ekspresi gen CDR1, ERG2 dan RTA3 baik pada C. albicans strain resisten dan sensitif, strain C. albicans resisten dan sensitif isolat Indonesia yang dikaji adalah satu spesies.

The aimed of this study was to compare the difference of CDR1, ERG2 and RTA3 gene expression between Candida albicans sensitive and resistant fluconazole strain Indonesian isolate, to identify the effect of Cyclosporine A (CY A) to those three genes for either the sensitive or resistant strain and to analyze the phylogenetic relationships between those two strains based on 18S rRNA gene sequences. The method used in gene expression analysis was a combination between RT-PCR and Densitometry, while to analyze phylogenetic relationships between those strains were using 18S rRNA sequence analysis method with the assistance of Bioedit and MEGA 4.0.2 software. The result showed that gene expression of CDR1, ERG2 and RTA3 were different between sensitive and resistant strain. Resistant strain could not express those genes (negative), but sensitive strains strongly express most of genes (positive). The effect of CY A was causing the resistant strain R781 and sensitive strains S300, S662, S966, S1023 and S1034 strongly expressed CDR1, ERG2 and RTA3 genes. The phylogenetic analysis result showed that all strains used in this research had the same 18S rRNA genes sequence. From this study we can concluded that CDR1, ERG2 and RTA3 were expressed differently between sensitive and resistant strains, the CY A effect was to increase CDR1, ERG2 and RTA3 genes expression either for sensitive or resistant strain, and all C. albicans strains used in this research were in one species.

Kata Kunci : CY A,CDR1,ERG2,RTA3,C albicans