Skripsi

Pluripotency level identification of breast cancer cell fractions separated by Magnetic Activated Cell Sorting through the analysis of cMyc gene expression = Identifikasi Tingkat Pluripotensi dari Fraksi Sel Punca Kanker Payudara yang Telah Dipisahkan dengan Magnetic Activated Cell Sorting (MACS) Dilihat dari Ekspresi Gen C-Myc.

Background: According to WHO, breast cancer has the highest incidence among women. Breast cancer occurs when there is an uncontrolled growth of abnormal cells formed in tissues of the breast, triggered by the presence of initiating cells called Cancer Stem Cells (CSC). The invasive properties possessed by breast CSCs are highly associated with the pluripotency level of the cells. The pluripotency level is highly related to the genes that are expressed. In this research, c-Myc gene expression is observed to determine the pluripotency level of the breast CSC fractions separated using Magnetic Activated Cell Sorting (MACS). Method: mRNA extracted from 11 breast cancer stem cell fractions that have been separated using Magnetic Activated Cell Sorting (MACS) are amplified using one step real time RT-PCR with SYBR Green (Bioneer®). Relative expression of c-Myc mRNA expression is analyzed through the calculation of cycle threshold (Ct) values using Livak method and normalized towards 18S rRNA as housekeeping gene to determine the pluripotency level of the breast CSC fractions. Results: Based on the results of the experiment, high level expression of c-Myc is present in CD24-/44- fraction sample, whereas low level c-Myc expression is present in CD24-/44+. Conclusions: C-Myc gene is expressed in all of the breast CSC fractions. Based on c-Myc expression analysis, high pluripotency level was found in CD24-/44- compared to CD24-/44+.
Keywords: Magnetic Activated Cell Sorting, c-Myc, CD24, CD44, pluripotency.


Background: Menurut WHO, kanker payudara memiliki angka kejadian paling tinggi diantara wanita. Kanker payudara disebabkan oleh pertumbuhan yang tidak terkendali dari sel-sel abnormal yang terbentuk di jaringan payudara, dipicu oleh kehadiran sel punca (Cancer Stem Cell). Sifat invasif yang dimiliki sel punca payudara terkait erat dengan pluripotensi sel tersebut. Pluripotensi dari sel sangat terkait dengan gen yang diekspresikan. Dalam penelitian ini, ekspresi gen c-Myc diamati untuk menentukan tingkat pluripotensi sampel fraksi sel punca kanker payudara yang dipisahkan dengan teknik Magnetic Activated Cell Sorting (MACS). Method: mRNA diperoleh dari 11 sampel sel punca kanker payudara yang difraksinasi menggunakan MACS. Ekspresi c-Myc di dalam fraksi-fraksi sel tersebut dianalisis menggunakan one step real time RT-PCR dengan SYBR Green (Bioneer®) dan elektroforesis. Results: Berdasarkan hasil eksperimen, ekspresi tingkat tinggi c-Myc hadir dalam fraksi sel CD24-/44-, sedangkan ekspresi tingkat rendah gen c-Myc ditemukan dalam fraksi sel CD24-/44+. Conclusions: Gen c-Myc diekspresikan di dalam semua fraksi sel punca kanker payudara. Melihat dari ekspresi gen c-Myc, level pluripotensi yang lebih tinggi dapat ditemukan di dalam fraksi sel CD24-/44- dibandingkan CD24-/44+.
Keywords: Magnetic Activated Cell Sorting, c-Myc, CD24, CD44, pluripotency