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"Kanker payudara menjadi penyebab kematian utama akibat kanker pada wanita. Metastasis dan kekambuhan menjadi faktor penyebab utama kematian akibat kanker. Metastasis menyebabkan sel tumor menginvasi dan menyebar melalui pembuluh darah menuju organ tubuh lain dan resistensi disebabkan karena sel punca yang memiliki kemampuan untuk self-renewal. Gen EpCAM dan CD44 dilaporkan memiliki kaitan dengan kepuncaan sel kanker. Sampai saat ini, pengembangan pengobatan kanker payudara masih terus dilakukan. Penggunaan kultur primer dalam studi in vitro terus dikembangkan karena hasil kultur primer homogen dengan lingkungan kanker primer. Optimasi kultur primer masih perlu dikembangkan. Selain itu, untuk melihat kepuncaan sel kanker diperlukan studi ekspresi gen terkait sel punca, yaitu EpCAM dan CD44. Penelitian ini bertujuan untuk mengoptimasi kultur primer kanker payudara dan mendeteksi sel punca menggunakan gen EpCAM dan CD44. Sampel kanker payudara didapatkan dari 10 pasien RS Cipto Mangunkusumo. Sampel yang digunakan adalah sampel high proliferative dan low proliferative. Metode kultur primer yang digunakan adalah metode enzimatis dan eksplan. Pengamatan kultur sel dilakukan selama 30 hari. Pada pengamatan molekuler, jaringan asal kanker dan sel hasil kultur primer digunakan untuk melihat ekspresi gen menggunakan metode qPCR. Hasil yang diperoleh menunjukkan bahwa metode yang berhasil untuk menumbuhkan sel kanker payudara adalah metode eksplan dan karakteristik sampel high proliferative. Sel sferoid (3D) didapatkan pada kultur kanker payudara. Hasil ekspresi gen menunjukkan ekspresi EpCAM dan CD44 tidak berbeda nyata (P>0,05) antara hasil kultur dan jaringan asal. Ekspresi gen yang tinggi diketahui berkorelasi dengan kehadiran sel punca

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Breast cancer is the leading cause of death from cancer in women. Metastases and relapses are the main contributing factors to death from cancer. Metastases cause tumor cells to invade and spread through blood vessels to other organs of the body and resistance is caused due to stem cells having the ability to self-renew. The EpCAM and CD44 genes are reported to be associated with cancer cell stemness. To date, the development of breast cancer treatment is still being developed. The use of primary culture in in vitro studies continues to be developed because the results of the primary culture are homogeneous with the primary cancer environment. However, optimization of primary culture is still required to be developed. In addition, to see the cancer stemness, studies of stem cell-related gene expression are needed, namely EpCAM and CD44. This study aims to optimize the primary culture of breast cancer and detect stem cells using the EpCAM and CD44 genes. Breast cancer samples were obtained from 10 patients at Cipto Mangunkusumo Hospital. The samples used were high proliferative and low proliferative samples. The primary culture methods used were enzymatic and explanatory methods. Observation of cell cultures was carried out for 30 days. In molecular observations, cancer origin tissue and primary cultured cells were used to see gene expression using the qPCR method. The results obtained showed that the successful method for growing breast cancer cells is the explant method. Spheroid (3D) cells were obtained in breast cancer cultures. Gene expression results showed that EpCAM and CD44 expression did not differ significantly (P>0.05) between culture results and tissue origin. High gene expression is known to correlate with the presence of stem cells."

"Kanker kolorektal merupakan salah satu kanker yang memiliki angka penderita yang tinggi di dunia. EPCAM adalah salah satu gen yang diekspresikan pada karsinoma dan ditemukan dalam circulating tumour cell (CTC) yang umum digunakan sebagai penanda kanker. Peripheral blood mononuclear cell (PBMC) adalah sel darah perifer yang juga diduga mengekspresikan gen EPCAM, dan perlu diteliti lebih lanjut sebab kelimpahan PBMC di dalam tubuh lebih banyak dibandingkan CTC. Penelitian dilakukan dengan tujuan untuk mendeteksi ekspresi gen EPCAM pada CTC dan PBMC kanker kolorektal menggunakan metode semi-quantitative RT-PCR dan direct immunofluorescence sehingga potensi EPCAM sebagai biomarker untuk diagnosis kanker kolorektal diketahui. Isolat CTC dan PBMC yang berasal dari delapan sampel darah penderita kanker kolorektal diteliti melalui isolasi dan kuantifikasi RNA, kemudian dilanjutkan dengan amplifikasi cDNA, dan pewarnaan antibodi. Hasil penelitian menunjukkan tidak adanya ekspresi gen EPCAM pada CTC dan PBMC menggunakan metode semi-quantitative RT-PCR. Hasil pengamatan dengan metode direct immunofluorescence menunjukkan pada salah satu sampel PBMC terdapat protein yang belum dapat dipastikan sebagai EpCAM, namun pada CTC ekspresi gen EPCAM tidak terdeteksi. Berdasarkan hasil penelitian, dapat disimpulkan bahwa ekspresi gen EPCAM tidak terdeteksi pada CTC dan PBMC dengan metode semi-quantitative RT-PCR. Penelitian lebih lanjut diperlukan untuk mengkonfirmasi protein yang terdeteksi pada PBMC adalah EpCAM.

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Colorectal cancer is one of the most common cancers in the world. EPCAM is one of the genes expressed in carcinoma and found in circulating tumor cells (CTC) which is commonly used as a cancer marker. Peripheral blood mononuclear cell (PBMC) is a peripheral blood cell that is also thought to express the EPCAM gene and needs to be investigated further because PBMC is more abundant in the body than CTC. The research was conducted to detect the expression of the EPCAM gene in CTC and PBMC of colorectal cancer using semi-quantitative RT-PCR and direct immunofluorescence methods so that EPCAM potential as a biomarker for colorectal cancer diagnosis is known. CTC and PBMC isolated from eight blood samples of colorectal cancer patients were evaluated through RNA isolation and quantification, followed by cDNA amplification, and antibody staining. The results showed that EPCAM gene expression was not detected both in CTC and PBMC using the semi-quantitative RT-PCR method. Furthermore, the direct immunofluorescence method showed that EPCAM gene expression was not detected in CTC. However, a protein that has not been confirmed as EpCAM was detected in PBMC. Based on the results of the study, it can be concluded that the EPCAM gene expression was not detected both in CTC as well as in PBMC by semi-quantitative RT-PCR method. Further research is required to confirm that the protein detected in PBMC is EpCAM.CTC, direct immunofluorescence, ekspresi gen, EPCAM, kanker kolorektal, PBMC, semi-quantitative RT-PCR."