Induksi Kerusakan pada Sel Epitel (HeLa) oleh Bakteri Komensal Oral Streptococcus sanguinis

• Main Authors: Susilowati, Heni, Hutomo, Maria Margareta Suryani, Lenggogen, Puti
• Other Authors: Diansari, Viona
• Format: Book PeerReviewed
• Language: English; English
• Published: Fakultas Kedokteran UNSYIAH Darussalam Aceh 2013
• Subjects: Medical Bacteriology
• Online Access: https://repository.ugm.ac.id/139047/1/AsyiahDMIII%20abstract.pdf; https://repository.ugm.ac.id/139047/7/Asyiah%20DM%202014%20Full%20paper%20Prosiding%20Heni.pdf; https://repository.ugm.ac.id/139047/
• Institution: Universitas Gadjah Mada

Abstrak Bakteri komensal oral Streptococcus sanguinis sejauh ini diketahui sebagai spesies yang bertanggung-jawab pada kolonisasi awal bakteri pada permukaan gigi. Bakteri ini secara jelas berperan dalam patogenesis infective endocarditis dan dijumpai pada lesi jaringan periodontal; namun perannya dalam patogenesis infeksi oral masih belum diketahui dengan jelas. Penelitian terhadulu menunjukkan bahwa S. sanguinis mampu menginduksi apoptosis pada sel epitel HeLa, tetapi mekanisme kematian sel tersebut belum jelas. Tujuan penelitian ini adalah untuk mengkaji pengaruh bakteri S. sanguinis pada prubahan seluler dan molekuler nukleus sel HeLa. Sel HeLa (5x105 sel/well) dikultur dalam RPMI 1640 semalam sebelum stimulasi. Suspensi bakteri S sanguinis (107 sel/ml) ditambahkan pada kultur HeLa dan diinkubasi selama 24 dan 48 jam dalam medium tanpa antibiotik. Analisis morfologi sel HeLa dilakukan dengan menggunakan mikroskop fase kontras dan mikroskop fluoresen setelah pewarnaan nukleus Hoechst 33342. Keutuhan DNA pada kelompok kontrol dan stimulasi bakteri diamaati setelah elektroforesis gel agarose. Profil DNA setiap sampel diamati menggunakan UV transiluminator. Doxorubicin (0,5625 mg/ml) digunakan sebagai kontrol positif induksi apoptosis. Hasil penelitian ini menunjukkan bahwa S. sanguinis menyebabkan perubahan morfologi sel, ditandai dengan berkurangnya ukuran sel seperti tampak pada mikroskop fase kontras serta peningkatan intensitas serapan fluoresen nukelus. Degradasi DNA terjadi pada sel yang terpapar S. sanguinis. Disimpulkan bahwa S. sanguinis mampu menginduksi kerusakan pada sel epitel HeLa. Kata kunci: Streptococcus sanguinis, perubahan morfologi sel, kerusakan DNA Abstract An oral commensal bacterium Streptococcus sanguinis is known as species which is responsible for early bacterial colonization within dental plaque. This bacterium has been implicated clearly with the pathogensis of infective endocarditis and some periodontal lession; however, it’s role in the patogenesis of oral infection still unknown. In previous study it was found that S. sanguinis induced apoptotic cell death on epithelium cell line, HeLa cells, but the mechanism still needs to be clarified. Therefore, the aim of this study was to investigate the effect of S. sanguinis whole cell bacteria to cellular and molecular change of HeLa cells nuclei. The cells (5x105 cells/well) were cultured in complete RPMI 1640 medium for overnight before stimulation. Streptococcus sanguinis bacterial suspension (107 cells/ml) was added to the culture and incubated for 24 and 48 hours in antibiotic-free culture medium. HeLa cells morphological analysis was performed under phase contrast and fluorescence microscope observation after Hoechst 33342 staining. In order to observe the integrity of DNA, HeLa whole cell DNA of untreated and bacteria-treated group were isolated addressed for agarose gel electrophoresis. The DNA profil of each sample was observed under UV transilluminator. An apopotic inducer Doxorubicin (0,5625 mg/ml) was used as a positive control in this study. The results demonstrated that S. sanguinis caused cell morphological change on HeLa cells indicated by decrease in cell size as shown on phasecontrast microscopy and increased fluorescence intensity of nucleus. A smear DNA degradation was observed after agarose gel electrophoresis of S. sanguinis-treated cells. It was concluded that S. sanguinis could induce cell damage on HeLa cells. Keywords: Streptococcus sanguinis, cell morphological change, DNA damage