Deteksi Natrium/Iodide Symporter (NIS) pada Galur Sel Kanker Payudara SKBR3 dengan Imunositofluoresens

Abstract

Galur sel SKBR3 adalah model kanker payudara positif human epidermal growth factor receptor2 (HER2). Pemberian kemoterapi memperlihatkan respons lengkap hanya pada 50% pasien kanker payudara dengan tipe positif HER2. Kemampuan jaringan tumor menangkap dan mengakumulasi iodium radioaktif dihubungkan dengan ekspresi natrium/iodide symporter (NIS). Tujuan penelitian ini adalah menilai ekspresi dan distribusi NIS pada galur sel SKBR3 serta menilai efek induksi epidermal growth factor (EGF) pada ekspresi NIS menggunakan imunositofluoresens-ISF. Penelitian ini dilakukan di Laboratorium Kultur Sel, Fakultas Kedokteran Universitas Padjadjaran (FKUP) mulai bulan September 2013 sampai dengan April 2014. Sel SKBR3 ditumbuhkan pada plat kultur dan ditunggu hingga konfluen 70%. Sel dibagi atas dua kelompok, yaitu kelompok yang diberi induksi dan kontrol. Induksi EGF diberikan dengan dosis 50 ng/mL. Pemeriksaan ISF menggunakan antibodi primer rabbit polyclonal antibody anti NIS dan antibodi sekunder goat anti rabbit IgG polyclonal antibody. Data hasil pengamatan dinilai secara semikuantitatif. Natrium/iodide symporter tampak terekspresi dan terdistribusi di sitoplasma. Sel yang diinduksi dengan EGF memperlihatkan peningkatan ekspresi NIS di sitoplasma dan distribusinya di membran sel secara bermakna. Sel SKBR3 mengekspresikan NIS yang terdapat di sitoplasma. Induksi EGF meningkatkan ekspresi NIS dan distribusinya di membran sel. Temuan ini dapat mengarah potensi kemampuan sel kanker payudara menangkap dan mengakumulasikan iodium radioaktif. [MKB. 2016;48(1):15–8]

Kata kunci: Ekspresi NIS , galur sel SKBR3, kanker payudara, imunositofluoresens

Detection of Natrium/Iodide Symporter (NIS) in SKBR-3 Breast Cancer Cell Line Using Immunocytofluoresence

Abstract

SKBR-3 cell line is a breast cancer model for human epidermal growth factor receptor2 (HER2) positive. Only 50% of patients of this type have fully responded to chemotherapy. Natrium iodide symporter expression correlates with the uptake and ability of cells to accumulate radioiodine. The aim of this study was to examine natrium/iodide symporter (NIS) expression and its distribution with and without epidermal growth factor (EGF) treatment using immunocytofluoresence (ICF). This study was conducted at the Cell Culture Laboratory, Faculty of Medicine, Universitas Padjadjaran from September 2013 to April 2014. SKBR3 cells were cultured until 70% confluent. Cells were then divided into two groups: treatment group and control group. The treatment group was treated with EGF 50 ng/mL. Cells were incubated with primary antibody rabbit polyclonal antibody anti-NIS, and then were followed with secondary-antibody goat polyclonal antibody to rabbit. Data from the observation were then assessed semi-quantitatively. Natrium/iodide symporter was seen to be expressed and distributed in the cytoplasm. Cells induced by EGF showed significant increase in NIS expression in cytoplasm and its distribution in cell membrane. It is concluded that the SKBR3 cells express NIS in cytoplasm and that EGF induction increases NIS expression and distribution in cell membrane. This finding leads to a potential ability of breast cancer cells to uptake and accumulate radioiodine. [MKB. 2016;48(1):15–8]

Key words: Breast cancer, cell line SKBR-3, immunocytofluoresence, NIS expression