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"Kanker payudara merupakan salah satu penyebab kematian utama secara global, dengan angka kematian yang terus meningkat, khususnya pada wanita. Kasus kematian kanker payudara pada umumnya terjadi karena metastasis yang dipengaruhi oleh faktor Epithelial-mesenchymal transition (EMT). Zinc finger E-box binding homeobox 1 (ZEB1) diketahui berperan dalam proses deregulasi EMT. Penggunaan jaringan asli dan kultur primer dari pasien kanker payudara memainkan peran penting dalam memeriksa perilaku kanker payudara, khususnya proses migrasi sel dan karakterisasi molekuler. Penelitian ini bertujuan untuk mengetahui potensi kultur eksplan dalam memprediksi kemampuan migrasi sel kanker payudara in vitro, serta analisis ekspresi gen ZEB1 dari penderita kanker payudara. Penelitian ini menggunakan jaringan dari penderita kanker payudara yang dikultur dengan metode eksplan dan diamati dibawah mikroskop, kemudian gen ZEB1 diisolasi dan dianalisis menggunakan qPCR. Hasil penelitian menunjukkan bahwa sel BC02 yang dikategorikan ganas berdasarkan nilai imunohistokimia dan patologi anatomi, membutuhkan waktu kurang dari tujuh hari untuk bermigrasi dari tumor primer, sedangkan BC01 yang dikategorikan jinak membutuhkan waktu 21 hari. Laju migrasi sel dari jaringan diperkirakan bergantung pada status keganasan jaringan. Ekspresi gen ZEB1 pada jaringan dan hasil kultur primer tidak berbeda nyata (p>0.05). Ekspresi ZEB1 pada S04 dan S09 yang dikategorikan sel ganas berdasarkan nilai imunohistokimia dan patologi anatomi berkorelasi positif dengan kemampuan migrasi berdasarkan tingkat keganasan sel kanker payudara. Penelitian ini menunjukkan bahwa kultur eksplan dapat digunakan untuk mempelajari karakteristik migrasi sel kanker. Selain itu, berdasarkan penelitian ini diketahui adanya hubungan ekspresi ZEB1 dengan tingkat keganasan sel kanker.

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Breast cancer is one of the leading causes of death globally, with cases of death increasing, especially in women. Cases of death in breast cancer occur due to metastases mediated by Epithelial-mesenchymal Transition (EMT) factors. Zinc finger E-box binding homeobox 1 (ZEB1) has been reported to play a role in the EMT deregulation process. The use of patient-derived primary cultures from breast cancer patients plays an important role in examining the behavior of breast cancer, in particular the process of cell migration and molecular characterization. This study aims to determine the potential of explant culture in predicting the migration ability of breast cancer cells in vitro, and molecular characterization by studying the expression of the ZEB1 gene in breast cancer patients. The results showed that BC02 cells took less than seven days to migrate from the primary tumor, while BC01 cells took 21 days. The rate of cell migration from the tissue was found to depend on the malignant status of the tissue. ZEB1 gene expression in tissue and primary culture were not significantly different (p>0.05). ZEB1 expression in S04 and S09 which were was positively correlated with migration ability based on the malignancy level of breast cancer cells. Furthemore, ZEB1 expression was found to be correlated with the grade of malignancy of breast cancer cells"

"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 payudara (KPD) merupakan kanker dengan jumlah insidensi dan mortalitas tertinggi pada wanita di dunia dan Indonesia pada tahun 2020. Usaha pencarian biomarka tambahan dilakukan untuk membantu deteksi dini dan evaluasi prognosis. Sebelumnya, jumlah salinan DNA Mitokondria (mtDNA-CN) dan panjang relatif telomer (RTL) dari darah perifer ditemukan berasosiasi dengan peningkatan risiko KPD. Keduanya dapat dipengaruhi oleh perubahan sistemik, seperti stres oksidatif. Penelitian ini bertujuan untuk menganalisis asosiasi mtDNA-CN dan RTL dengan KPD di Indonesia. Penelitian kasus-kontrol ini melibatkan 209 subjek kontrol dan 197 subjek kasus yang berasal dari rumah sakit di 5 daerah di Indonesia (Jakarta, Semarang, Pekanbaru, Makassar, Kupang). Teknik qPCR digunakan untuk mengamplifikasi gen referensi (B2M), mtDNA (MT-TL1), dan telomer. Rasio mtDNA-CN dan RTL dihitung berdasarkan hasil perbandingan terhadap B2M. Asosiasi mtDNA-CN dan RTL dengan risiko dan prognosis KPD dianalisis menggunakan uji regresi. Hasil penelitian menunjukkan bahwa mtDNA-CN berasosiasi positif dengan RTL (p<0,025). MtDNA-CN yang lebih banyak dan RTL yang lebih panjang, serta kombinasi ‘tinggi-tinggi’ dari keduanya ditemukan berasosiasi signifikan dengan peningkatan risiko KPD pada kelompok usia <48 tahun (p<0,025). Selain itu, mtDNA-CN dan RTL berasosiasi signifikan dengan beberapa karakteristik klinis patologis KPD. MtDNA-CN dan RTL berpotensi digunakan sebagai biomarka risiko dan prognosis KPD.

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In 2020, breast cancer has been the leading cause of cancer incidence and mortality among women globally, including in Indonesia. Additional biomarkers discovery were required for early detection and prognostic evaluation. Previously, the peripheral blood mitochondrial DNA copy number (mtDNA-CN) and relative telomere length (RTL) had been associated with elevated breast cancer risk. Both markers might be influenced by the change in systemic condition, such as oxidative stress. We aimed to investigate the associations between mtDNA-CN and RTL with breast cancer in Indonesia. A total of 209 controls and 197 cases from several hospitals in 5 locations in Indonesia (Jakarta, Semarang, Pekanbaru, Makassar, Kupang) were enrolled. The reference gene (B2M), mtDNA (MT-TL1), and telomeres were amplified using qPCR method. The mtDNA-CN and RTL ratio were calculated by comparing them to B2M and the associations were analyzed using regression test. The results showed a significant positive association between mtDNA-CN and RTL (p<0,025). Higher mtDNA-CN, higher RTL, and a ‘high-high’ combination were significantly associated with elevated breast cancer risk in group with age <48 (p<0,025). Both markers were also associated with several clinicopathological features. Therefore, mtDNA-CN and RTL might potentially be used as biomarkers for breast cancer risk and prognosis."

"Pendahuluan: Tingkat kematian pasien kanker payudara menempati posisis tertinggi dunia dan umum terjadi pada wanita. TNBC merupakan jenis kanker payudara yang memiliki prognosis buruk. Selain tidak memiliki reseptor hormonal (ER, PR, dan HER2), TNBC memiliki kemampuan yang tinggi dalam mempertahankan hidup dan menolak apoptosis. Hal ini berkaitan dengan tingginya ekspresi gen Survivin, sebagai protein IAP (inhibitor apoptosis protein) yang berperan dalam menghambat apoptosis dan meningkatkan proliferasi. Studi terbaru menunjukkan bahwa CRISPR/Cas9 merupakan pendekatan yang sesuai untuk mengedit gen yang berpeluang besar dalam terapi kanker.Bahan dan Metode: Sel TNBC BT549 yang telah ditransfeksi dengan Cas9 dan sgRNA. Analisis molekuler dimulai dengan analisis aktifitas pembelahan gen menggunakan (PCR), efisiensi pengeditan genom (Sanger Sequencing), ekspresi mRNA Survivin (qRT-PCR), dan analisis ekspresi protein Survivin (westernblot). Analisis fenotipe dilakukan dengan uji apoptosis (flowcytometry) dan proliferasi (tripan-blue exclusion assay). Analisis bioinformatika dilakukan dengan menganalisis struktur protein (PyMoL) dan analisis interaksi protein (Cytoscape).Hasil: CRISPR/Cas9 berhasil menghilangkan fungsi gen Survivin pada sel TNBC BT549. Penurunan ekspresi mRNA dan protein Survivin signifikan, peningkatan apoptosis dan penghambatan proliferasi pada sel TNBC BT549.Kesimpulan: Penelitian ini merupakan riset pertama kali yang berhasil membuktikan efek dari KO Survivin pada apoptosis dan proliferasi sel TNBC BT549.

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Introduction: Breast cancer has the highest mortality rate in the world and is most common in women. TNBC is a type of breast cancer with a poor prognosis. TNBC, in addition to lacking hormonal receptors (ER, PR, and HER2), has a high ability to maintain life and resist apoptosis. This is due to the high expression of the Survivin gene, an apoptotic inhibitor protein that plays a role in inhibiting apoptosis and increasing proliferation. Recent research has shown that CRISPR/Cas9 is a suitable approach for gene editing with great potential in cancer therapy.

Materials and Methods: Cas9 and sgRNA were transfected into BT549 TNBC cells. Molecular analysis with PCR, genome editing efficiency (Sanger Sequencing), Survivin mRNA expression (qRT-PCR), and protein expression analysis (westernblot). Phenotypic analysis were carried out by apoptosis (flowcytometry) and proliferation (trypan-blue exclusion assay). Protein structure were studied using (PyMoL) and protein interaction (Cytoscape).

Results: CRISPR/Cas9 successfully eliminated the function of the Survivin gene in BT549 TNBC cells. Significant reduction in Survivin mRNA and protein expression, increased apoptosis, and inhibition of proliferation in BT549 TNBC cells.

Conclusion: This study is the first to demonstrate the effect of Survivin knockout on apoptosis and proliferation of TNBC BT549 cells."

"Kanker payudara merupakan salah satu jenis kanker dengan kematian terbanyak kedua di dunia. Penyebab utama kematian penderita kanker payudara adalah metastasis sel kanker ke jaringan lain. Salah satu penanganan kanker payudara adalah pemberian pengobatan kemoterapi. Namun kemoterapi seringkali menimbulkan efek samping pada pasien. Salah satu jenis kemoterapi umum, Doxorubicin mampu meningkatkan resiko toksisitas jantung. Karakteristik kultur eksplan yang dapat mempertahankan kondisi sel secara in vivo dapat digunakan untuk memprediksi respons kemoterapi. Namun demikian, penggunaan kultur eksplan untuk memprediksi respon kemoterapi belum banyak diterapkan. Gen MYCN memiliki peran dalam mendorong keganasan sel kanker dan banyak terekspresi pada kanker payudara dengan prognosis buruk, sehingga dapat digunakan sebagai biomarker untuk memastikan respon antara jaringan asal dan kultur eksplan relatif sama. Oleh karena itu penelitian ini bertujuan untuk menganalisis ekspresi gen MYCN pada jaringan asal dan kultur eksplan serta menganalisis tingkat ekspresi gen MYCN pada kultur eksplan terhadap treatment Doxorubicin dengan metode one-step semi-kuantitatif Reverse Transcriptation - Polymerase Chain Reaction (RT-PCR). Perlakuan treatment dilakukan untuk mendukung kemampuan kultur eksplan dalam mempertahankan in vivo. Hasil yang diperoleh menunjukkan ekspresi relatif gen MYCN pada jaringan asal dan kultur eksplan relatif sama serta adanya ekspresi gen MYCN setelah pemberian treatment pada kultur eksplan. Hal tersebut menandakan kultur eksplan dapat mempertahankan ekspresi dari jaringan asalnya. Selain itu, adanya ekspresi gen MYCN setelah pemberian agen kemoterapi mengkonfirmasi bahwa kultur eksplan memiliki respons yang relatif sama setelah pemberian treatment sehingga dapat digunakan untuk memprediksi respon kemoterapi.

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Breast cancer is one type of cancer with the second most deaths in the world. The main cause of death for breast cancer patients is the metastasis of cancer cells to other tissues. One of the treatments for breast cancer is chemotherapy treatment. However, chemotherapy often causes side effects in patients. One of the common types of chemotherapy, Doxorubicin can increase the risk of cardiac toxicity. Characteristics of explant cultures that can maintain cell conditions in vivo can be used predicting chemotherapy response. However, the use of explant cultures to predict chemotherapy response has not been widely applied. The MYCN gene has a role in promoting cancer cell malignancy and is widely expressed in breast cancer with a poor prognosis, so it can be used as a biomarker to ensure that the response between the tissue of origin and explant cultures is relatively similar. Therefore, this study aims to analyze MYCN gene expression in the original tissue and explant culture and to analyze the expression level of the MYCN gene in explant culture against Doxorubicin treatment using a one-step semi-quantitative Reverse Transcriptation - Polymerase Chain Reaction (RT-PCR) method. The treatment was carried out to support the ability of the explant culture to maintain in vivo. The results obtained showed that the relative expression of the MYCN gene in the original tissue and the explant culture was relatively the same as well as the expression of the MYCN gene after the treatment was given to the explant culture. This indicates that the explant culture can maintain the expression of the original tissue. In addition, the presence of MYCN gene expression after administration of chemotherapeutic agents confirmed that explant cultures had relatively the same response after treatment, so that they could be used to predict chemotherapy responses."

"Kanker payudara merupakan penyakit kanker yang berkembang dari jaringan epitel duktus dan kelenjar susu dalam payudara. Kanker payudara merupakan kanker dengan prevalensi tertinggi di dunia dan telah mengalami kenaikan jumlah kasus yang signifikan (16,6%) per tahun 2020 di Indonesia. Pengobatan dengan kemoterapi telah umum dilakukan namun dilaporkan memiliki banyak efek samping. Oleh karena itu, perlu dilakukan prediksi kemosensitivitas pada pasien sebelum obat diberikan. Prediksi kemosensitivitas dapat dilakukan dengan menggunakan kultur primer metode eksplan sebagai patient-derived model in vitro yang menggunakan sel primer langsung dari pasien dan dapat memprediksi respons klinis terhadap obat. Adanya ekspresi relatif gen c-Myc selaku oncogene addiction gene dapat menjadi biomarker potensial pada kultur eksplan setelah diberi perlakuan Doxorubicin. Pada penelitian ini, ekspresi c-Myc pada jaringan asal dan kultur eksplan serta pada kultur eksplan sebelum dan sesudah perlakuan diukur menggunakan metode semi kuantitatif RT-PCR dengan tahapan isolasi RNA, kuantifikasi, amplifikasi, dan visualisasi dengan elektroforesis. Kultur eksplan yang berhasil ditumbuhkan adalah sampel luminal A dan B. Sampel A menunjukkan migrasi sel hingga konfluensi 50%. Sementara uji viabilitas pada hasil kultur sampel B menunjukkan persentase viabilitas kultur non-treatment 60% dan treatment dosis 0,68 μM 41%. Hasil visualisasi elektroforesis tidak menunjukkan adanya ekspresi c-Myc di seluruh sampel.

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Breast cancer is a malignant tumor formacy on ductus epithelial tissue and mammary gland of the breast. The prevalence number of breast cancer is currently leading among other cancers in the world and significantly increasing especially in Indonesia (16,6% new cases in 2020). Treatment using chemotherapy agents is the most common option for patients but has many side effects. Chemosensitivity prediction using an in vitro model is recommended for more personalized treatment, and can be performed using primary culture (explant method) as an in vitro patient-derived model that uses primary cells directly from the patient and can predict pathological response to chemotherapy agents. c-Myc gene as an oncogene addiction gene can be a potential biomarker of explant cultures after being treated by Doxorubicin. In this study, c-Myc expression in fresh tissue and explant culture, also in culture prior to and after treatment, was measured using a semi quantitative RT-PCR method involving the process of RNA isolation, quantification, amplification, and visualization using electrophoresis. Luminal A and Luminal B samples of explant culture were successfully grown and harvested. A sample showed cell migration up to 50% confluency. Meanwhile, the viability test on the results of the B culture showed the viability percentage of non-treatment culture was 60% and 0,68 μM treatment dose culture 41%. The results of electrophoretic visualization consistently showed downregulation of c-Myc in the form of a very low expression in all samples."

"Kanker payudara adalah jenis kanker yang memiliki kasus baru terbanyak dan penyebab kematian tertinggi di kalangan wanita. Hal tersebut menjadikan pemahaman mengenai konsep pengobatan presisi (precision medicine) perlu dikembangkan, salah satunya dengan penggunaan kultur primer dari jaringan kanker pasien. Epidermal growth factor receptor (EGFR) telah dilaporkan umum digunakan sebagai marker prognostik kanker payudara melalui deteksi protein menggunakan metode immunohistokimia (IHK). Namun, metode tersebut memiliki kekurangan dalam menjadi acuan penentuan terapi adjuvant. Penggunaan kultur primer sebagai pengganti cell lines dalam penelitian kanker perlu terus dikembangkan karena lebih mewakili karakteristik fenotipe dan genotipe dari jaringan kanker in vivo. Oleh sebab itu, penelitian ini bertujuan untuk mengetahui tingkat ekspresi relatif gen EGFR pada sampel kultur primer dan jaringan asalnya serta mengetahui perbandingan ekspresi relatif gen EGFR pada sampel jaringan jinak dan jaringan ganas kanker payudara. Metode yang digunakan, yaitu melalui deteksi mRNA gen EGFR dengan semi kuantitatif RT-PCR pada dua pasien yang mewakili jaringan jinak dan ganas. Hasil penelitian menunjukkan tidak terdapat perbedaan signifikan antara ekspresi EGFR pada kultur primer dan jaringan asal, serta ekspresi EGFR pada jaringan ganas lebih tinggi dibandingkan dengan jaringan jinak. Berdasarkan hasil yang diperoleh, dapat disimpulkan bahwa kultur primer dapat dijadikan model alternatif dalam penelitian kanker, serta EGFR dapat dijadikan marker potensial dalam menentukan tingkat agresivitas kanker payudara.

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Breast cancer is a type of cancer that has a highest rate of incidence as well as mortality among women. This makes the understanding of the concept of precision medicine need to be continuously developed, in wich one of the methods is through using primary cultures from patient's tissue. It has been reported that epidermal growth factor receptor (EGFR) is commonly used as a prognostic marker of breast cancer through protein detection using the Immunohistochemical (IHC) method. However, this method has shortcomings in being a reference for determining adjuvant therapy. The use of primary culture as a subtitute cell lines in cancer research needs to be developed due to its more representative of the phenotype and genotype characteristics of cancer tissue in vivo. Therefore, this study aims to determine the relative expression level of the EGFR gene in primary culture sample and its tissue origin as well as comparing the relative expression of the EGFR gene in samples of benign tissue and malignant tissue of breast cancer. The method used is the detection of EGFR gene mRNA with semi-quantitative RT-PCR in two patients representing benign and malignant tissues. The results showed that there was no significant difference between EGFR expression in primary culture and tissue of origin, and EGFR expression in malignant tissue was higher than in benign tissue. Hence, it can be concluded that primary culture can be used as an alternative model in cancer research, and EGFR can be used as a potential marker in determining aggressiveness level of breast cancer."

"Gen SOX2 telah dilaporkan memegang peranan penting dalam menginduksi sel punca progenitor dari sel fibroblast manusia dewasa. Peningkatan ekspresi gen SOX2 berkorelasi dengan peningkatan tingkat keparahan kanker payudara. Namun, bagaimana SOX2 memiliki sifat onkogenik belum diketahui secara pasti. Penelitian ini bertujuan untuk mendapatkan informasi mengenai ekspresi gen SOX2 pada sel kanker payudara CD44+/CD24-dan CD44-/CD24-yang di ko-kultur dengan Mouse Embryonic Fibroblast (MEF) berdasarkan waktu pengkulturan sel sebagai upaya untuk mempelajari ekspresi gen dan sifat dari sel punca kanker payudara pada sel kanker payudara dari pasien kanker payudara wanita Indonesia. Tingginya ekspresi gen SOX2 diasumsikan dapat menjadi indikasi untuk menentukan kondisi optimum pada kultur sel kanker payudara. Level RNA gen SOX2 diukur dengan menggunakan reverse transcription-polymerase chain reaction (RT-PCR) dan dinormalisasi dengan menggunakan housekeeping gene PUM1 sebagai kontrol dalam. Hasil menunjukkan bahwa ekspresi gen SOX2 tertinggi di hari ketiga pada kultur sel punca kanker (CD44+/CD24-), demikian pula dengan kultur sel non punca (CD44-/CD24-), dan di hari pertama pada kultur sel kanker payudara (CD44/CD24).

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SOX2 gene has been reported to play an important role in inducing stem cell progenitor cells from adult human fibroblasts. Increase in SOX2 gene expression known to correlate with the increase of breast cancer severity. However, the oncogenic properties of SOX2 has not been confirmed yet. This research aimed to obtain information about the level of SOX2 gene expression of breast cancer cells CD44+/CD24-dan CD44-/CD24-in co-culture with Mouse Embryonic Fibroblast (MEF) based on time of cell culture, in an attempt to study the gene expression and the nature of cancer stem cells in breast cancer cell in Indonesian female breast cancer patient.

High SOX2 gene expression was assumed as the indication of the optimum culture condition of breast cancer cell. SOX2 gene RNA level was measured performing reverse transcription-polymerase chain reaction (RT-PCR) and normalized using the housekeeping gene PUM1 as an internal control. Results showed that the highest SOX2 gene expression was found in day 3 for cancer stem cell cultures (CD44+/CD24-) as well as for non cancer stem cell cultures (CD44-/CD24-), and in day 1 for breast cancer cell cultures (CD44/CD24). "

"[Latar Belakang: Kanker payudara masih merupakan kanker yang paling umumpada wanita. Identifikasi sel punca kanker payudara sangat penting dalammemberantas penyakit ini dari akarnya. Beberapa riset telah mengisolasi sel puncakanker payudara berdasarkan protein membran sel CD24/CD44 dan menemukansel punca kanker payudara pada sel CD24-/CD44+ yang menunjukkan sifatpluripotensi. Namun, beberapa riset lainnya menemukan CD24-/CD44+ tidakditemukan pada seluruh tipe kanker payudara, dan tidak selalu berhubungandengan perkembangan tumor. Maka dari itu, tingkat pluripotensi dari sel tersebutmasih diperdebatkan. Dalam riset ini, sifat pluripotensi sel punca kanker payudaradinilai berdasarkan ekspresi gen SOX2 yang merupakan gen untuk sifatkepuncaan dimana gen ini dapat mendorong pembelahan sel dan invasi.Metode: Sampel diambil dari situs primer kanker payudara dan difraksinasimelalui pemisahan sel magnetik. RT-qPCR dan elektroforesis digunakan untukmempelajari tingkat ekspresi gen SOX2 antara fraksi-fraksi sel punca kankerpayudara.Hasil: Kami berhasil memisahkan sel pluripoten dari spesimen klinis kankerpayudara. Fraksi CD24-/CD44- menunjukkan ekspresi gen SOX2 yang lebihtinggi secara signifikan dibanding CD24-/CD44+. Setelah melewati proses ultralowattachment, CD24-/CD44+ menunjukkan peningkatan ekspresi gen SOX2walaupun lebih rendah dari CD24-/CD44-.Kesimpulan: Pluripotensi yang tinggi, berdasarkan tingkat ekspresi gen SOX2,ditemukan pada fraksi CD24-/CD44-. Tingkat pluripotensi fraksi CD24-/CD44+lebih rendah dibandingkan fraksi CD24-/CD44-.;Background: Breast cancer remains as the most prevalent cancer in women.Identification of breast cancer stem cell (CSC) is crucial in eradicating the diseasefrom its root. Multiple research has isolated breast CSC based on CD24/CD44surface marker and discovered that CD24+/CD44- fraction indicates stemness andpluripotent characteristics. However, it was also found that CD24+/CD44- breastCSC is not present in all breast cancer types, and not always associated withtumor progression. Therefore, its pluripotency level remains debatable. In thisresearch, pluripotency of breast CSCs was assessed. Pluripotency was determinedbased on SOX2 gene expression, a gene responsible for stem-like properties,which can drive cellular proliferation and invasion.Method: The samples were taken from primary site of breast cancer andfractionated through magnetic cell sorting. RT-qPCR with subsequentelectrophoresis was used to study the expression level of SOX2 gene amongbreast CSC fractions.Results: We managed to separate the pluripotent cells from the bulk clinicalspecimen. CSC subset CD24-/CD44- showed a significantly higher SOX2expression in comparison to CD24-/CD44+. Following ultra-low attachment,CD24-/CD44+ showed an increase in SOX2 expression level although still lowerthan CD24-/CD44-.Conclusions: A high pluripotency based on SOX2 gene expression level wasfound in fraction CD24-/CD44-. The pluripotency level of fraction CD24-/CD44+was lower in comparison to fraction CD24-/CD44-., Background: Breast cancer remains as the most prevalent cancer in women.Identification of breast cancer stem cell (CSC) is crucial in eradicating the diseasefrom its root. Multiple research has isolated breast CSC based on CD24/CD44surface marker and discovered that CD24+/CD44- fraction indicates stemness andpluripotent characteristics. However, it was also found that CD24+/CD44- breastCSC is not present in all breast cancer types, and not always associated withtumor progression. Therefore, its pluripotency level remains debatable. In thisresearch, pluripotency of breast CSCs was assessed. Pluripotency was determinedbased on SOX2 gene expression, a gene responsible for stem-like properties,which can drive cellular proliferation and invasion.Method: The samples were taken from primary site of breast cancer andfractionated through magnetic cell sorting. RT-qPCR with subsequentelectrophoresis was used to study the expression level of SOX2 gene amongbreast CSC fractions.Results: We managed to separate the pluripotent cells from the bulk clinicalspecimen. CSC subset CD24-/CD44- showed a significantly higher SOX2expression in comparison to CD24-/CD44+. Following ultra-low attachment,CD24-/CD44+ showed an increase in SOX2 expression level although still lowerthan CD24-/CD44-.Conclusions: A high pluripotency based on SOX2 gene expression level wasfound in fraction CD24-/CD44-. The pluripotency level of fraction CD24-/CD44+was lower in comparison to fraction CD24-/CD44-.]"

"Latar Belakang: pulpa memiliki sifat low-compliance yang memengaruhi proses regenerasinya. Tujuan: menganalisis potensi Platelet- Rich Plasma (PRP) Eksosom terhadap regenerasi pulpa gigi secara in-vitro viabilitas sel, aktivitas migrasi, dan ekspresi Vascular Endothelial Growth Factor-A (VEGF-A) hDPSCs. Metodologi: hDPSC sembilan gigi molar tiga dikultur dengan metode enzyme digestion (ED) yang dipanen pada P3 dan P4. Kemudian dikultur di dalam enam media, yaitu: Dulbecco's Modified Eagle Medium (DMEM) dan 10% PRP sebagai kelompok kontrol, dan 0,5%, 1%, dan 5% eksosom dari PRP. Semua kelompok memiliki tiga rangkap biologis (Triplo). Uji viabilitas sel dievaluasi dengan MTT assay, aktivitas migrasi sel dengan Scratch Assay dan Transwell Migration Assay, dan ekspresi VEGF-A dengan Enzyme-Linked Lmmunosorbent Assay (ELISA). Analisis data dilakukan dengan uji One Way ANOVA (p <0,05) serta uji Kruskal-Wallis dan post hoc Mann-Whitney (p <0,05). Hasil: viabilitas hDPSCs tertinggi pada 24, 48 dan 72 jam observasi pada kelompok Eksosom dari PRP 5% (p <0,05). Eksosom dari PRP 5% menunjukkan aktivitas migrasi yang lebih tinggi dibandingkan dengan kelompok lain, meskipun terdapat perbedaan tidak bermakna dengan kontrol PRP 10% (p >0,05). Ekspresi VEGF-A hDPSCs tertinggi terdapat pada kelompok PRP Eksosom 5% pada 72 jam observasi. Kesimpulan: eksosom dari PRP 5% berpotensi menginduksi regenerasi pupa gigi manusia.

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Background: pulp has low-compliance properties that affect its regeneration process. Objective: to analyze the potential of Platelet-Rich Plasma (PRP) exosomes on the regeneration of dental pulp by in-vitro evaluation of cell viability, migration activity, and expression of Vascular Endothelial Growth Factor-A (VEGF-A) hDPSCs. Methodology: hDPSCs of nine third molars cultured by enzyme digestion (ED) method were harvested at P3 and P4. Then cultured in six media, Dulbecco's Modified Eagle Medium (DMEM) and 10% PRP as a control group, and 0.5%, 1%, and 5% exosomes of PRP. All groups had a biological triple (Triplo). Cell viability assay was evaluated by MTT assay, cell migration activity by Scratch Assay and Transwell Migration Assay, and VEGF-A expression by Enzyme-Linked Lmmunosorbent Assay (ELISA). Data analysis was performed using One Way ANOVA (p < 0.05) and Kruskal-Wallis and post hoc Mann-Whitney tests (p < 0.05). Results: The viability of hDPSCs was highest at 24, 48 and 72 hours of observation in the Exosomes group of 5% PRP (p < 0.05). Exosomes from 5% PRP showed higher migratory activity compared to other groups, although there was no significant difference with 10% PRP control (p > 0.05). The highest expression of VEGF-A hDPSCs was found in the 5% PRP Exosomes group at 72 hours of observation. Conclusion: exosomes of 5% PRP have the potential to induce the regeneration of human dental pulp."