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"ABSTRACTSel tumor adalah sel yang terbentuk akibat kegagalan beberapa protein dalam mengatur siklus sel. Protein TP53 berperan penting dalam mengatur siklus sel, khususnya dalam menekan perkembangan sel tumor. Perubahan pada gen TP53 ditemukan dalam lebih dari setengah kasus tumor pada manusia. Protein lain yang berhubungan dengan protein TP53 juga ditemukan terlibat dalam proses pembentukan kanker. Analisis interaksi protein TP53 dengan melakukan clustering jaringan interaksi protein (PPI) TP53 adalah hal penting dalam membantu mengatasi sel tumor. Jaringan PPI dinyatakan sebagai graf dengan protein dan interaksinya masing-masing sebagai simpul dan busur pada graf. Spectral clustering adalah metode graph clustering yang menggunakan eigenvector dari matriks Laplacian.

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ABSTRACTFuzzy random walk adalah metode fuzzy clustering yang menggunakan probabilitas transisi dari random walk pada data. Dua metode tersebut akan digabungkan dan diimplementasikan pada penelitian ini. Menggunakan data PPI protein TP53 dari STRING database, didapat gabungan kedua metode tersebut mampu menghasilkan cluster yang fuzzy dan robust di mana setiap cluster dapat menjelaskan bagian tertentu dari fungsi protein TP53. Tumor cell is formed as a result of malfunctioning of some proteins that regulates the cell cycle. TP53 protein plays an important role in managing cell cycle, especially in tumor cell suppression. An alteration of TP53 gene is found in more than half cases of human tumor. Moreover, TP53-related proteins are also found involved in the carcinogenesis process. Therefore, it is important to analyze the interactions of TP53 protein by clustering protein-protein interactions (PPI) network of TP53. PPI networks are usually represented as a graph network with proteins and interactions as vertices and edges respectively. Spectral Clustering is a graph clustering algorithm based on eigenvector of the graph Laplacian. Fuzzy Random Walk is a fuzzy clustering method based on transition probability from a random walk on a dataset. In this paper, we combine both Spectral Clustering and Fuzzy Random Walk. Using PPI datasets of TP53 obtained from the STRING database, we found the combined algorithm is proven to produce both robust and fuzzy clusters with each cluster explains one of TP53 proteins functionality."

"Pembentukan dan perkembangbiakan sel tumor terjadi jika protein khusus yang mengatur pembelahan sel mengalami perubahan fungsi, ekspresi gen atau hilang keduanya. Salah satu protein penekan tumor yang berperan dalam pengendalian siklus sel adalah protein TP53. Pada sebagian besar perubahan genetik dalam tumor, baik delesi atau mutasi pada lebih dari 50% kanker pada manusia, ditemukan mutan TP53 yang merupakan faktor beresiko tinggi terhadap kanker. Oleh karena itu, penting untuk melakukan studi tentang pengelompokan interaksi protein-protein TP53. Interaksi protein secara umum disajikan dalam jaringan graf (graph network) dengan protein sebagai simpul dan interaksinya sebagai busur. Algoritma Markov Clustering (MCL) adalah satu metode graph clustering yang dibuat berdasarkan simulasi dari flow stokastik pada suatu graf. Dalam skripsi ini, dibahas mengenai implementasi algoritma MCL pada data interaksi protein-protein TP53 dengan menggunakan bahasa pemrograman Python. Algoritma MCL terdiri dari tiga operasi utama yaitu ekspansi, penggelembungan, dan pemotongan. Selanjutnya, dilakukan analisis hasil clustering dari simulasi algoritma MCL dengan menggunakan parameter ekspansi, penggelembungan dan faktor pengali yang berbeda-beda. Berdasarkan analisis hasil clustering yang dilakukan, algoritma MCL terbukti menghasilkan robust cluster dengan protein TP53 sebagai pusat cluster untuk setiap hasil clustering.

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The formation and proliferation of tumor cells occurs if a special protein that regulates cell division changing the function, gene expression or lost both. One of the tumor suppressor protein that plays a role in controlling the cell cycle is the TP53 protein. In most of the genetic changes in the tumor, either deletions or mutations in more than 50% of human cancers, it found that mutant of TP53 is a high risk factor for cancer. Therefore, it is important to conduct studies on protein-protein interactions clustering of TP53. Protein interactions are generally presented in the graph network with proteins as nodes and interactions as edges. Markov Clustering (MCL) algorithm is a graph clustering method which is based on a simulation of stochastic flow on a graph. This minithesis discussed about the implementation of the MCL process on protein-protein interaction of TP53 data using the Python programming language. MCL algorithm consists of three main operations: expansion, inflation, and prune. Furthermore, the clustering simulation is using the different parameter of expansion, inflation and the multiplier factor. Based on the analysis of the clustering results, MCL algorithm is proven to produce robust cluster with TP53 protein as a centroid for each clustering results."

"ABSTRAK Latar belakang: Fibroadenoma dan tumor filodes jinak merupakan tumorfibroepitelial dengan gambaran histopatologik yang tumpang tindih. Saat inibanyak pengambilan jaringan tumor payudara secara core biopsy, termasuk padatumor fibroepitelial. Jumlah jaringan yang sedikit dan gambaran histopatologikyang tumpang tindih sering menyulitkan Dokter Spesialis Patologi Anatomikdalam menentukan diagnosis fibroadenoma dan tumor filodes jinak. Penelitian inibertujuan untuk mengetahui gambaran histopatologik apa saja yang bermaknauntuk mendiagnosis fibroadenoma dan tumor filodes jinak dan untuk mengujiapakah diagnosis fibroadenoma dan tumor filodes jinak pada core biopsy denganmenggunakan sistem skoring lebih baik dibandingan tanpa skoring.Bahan dan cara: Penelitian ini merupakan suatu uji diagnostik. 57 kasusfibroadenoma dan tumor filodes jinak yang memiliki slaid core biopsy danmastektomi/lumpektomi/eksisi dinilai ulang tanpa sistem skoring danmenggunakan skoring. Gambaran histopatologik yang dinilai pada sistem skoringadalah selularitas stroma, atipia inti, fragmentasi jaringan, infiltrasi lemak, mitosisdan heterogenitas stroma. Kemudian dilakukan analisis statistik, uji diagnostikdan uji kappa.Hasil: Selularitas stroma, heterogenitas stroma dan fragmentasi jaringan lebihsering ditemukan pada tumor filodes jinak dan berbeda bermakna (p=0,001;p=0,000; p=0,021). Spesifisitas pada sistem skoring meningkat sebesar 17,9%.Nilai duga positif dan nilai duga negatif pada sistem skoring meningkat sebesar11,9% dan 5,1%. Area under curve (AUC) meningkat 8,9%. Uji Cohen?s kappaantara diagnosis core biopsy tanpa dan dengan skoring bernilai rendah (0,545).Kesimpulan: Adanya peningkatan spesifisitas, nilai duga positif dan AUCmenunjukkan bahwa penilaian core biopsy sistem skoring lebih baikdibandingkan tanpa skoring dan dapat menjadi acuan untuk diagnosis fibroadenoma dan tumor filodes jinak.ABSTRACT Background: Fibroadenoma and benign phyllodes tumor are kinds of fibroepithelial tumor which have overlapping histopathological features. Recently, core biopsy is commonly performed to determine breast tumor, including fibroepithelial tumor. Small amount of tissue and overlapped histopathological features often complicate the Pathologist in diagnosing both. This study aims to describe the histopathological appearance which needed to diagnose fibroadenoma and benign phyllodes tumor and to verify if the diagnosis of fibroadenoma and benign phyllodes tumor in core biopsy using scoring system is more accurate than without scoring system.Method: This study was a diagnostic test, in which 57 cases of fibroadenoma and benign phyllodes tumor which had undergone core biopsy and mastectomy/excision were re-assessed using and without using scoring system. Histopathologic features which assessed using scoring system were stromal cellularity, nuclear atypia, tissue fragmentation, fat infiltration, mitotic figure, stromal heterogeneity. Analytical statistic, diagnostic test, accuracy test and Kappa test were done.Results: Stromal cellularity, stromal heterogeneity and tissue fragmentation mostly found in benign phyllodes tumor and significantly different (p=0,001; p=0,000; p=0,021).There were significant differences between stromal cellularity (p=0,001), stromal heterogeneity (p=0,000), and tissue fragmentation (p=0,021) in diagnosis of benign phyllodes tumor. Specificity in scoring system increased by17,9 %. Positive predictive value, negative predictive value and accuracy increased in scoring system (11,9% and 5,1%). Area under curve (AUC) increased by 8,9%. Cohen's Kappa test between core biopsy diagnosis without using and using scoring system had low result(0,545). Conclusion: The increasing of specificity, positive predictive value, accuracy and AUC proved that core biopsy with scoring system is more accurate than without scoring. This can be used as reference to diagnose fibroadenoma and benign phyllodes tumor.;Background: Fibroadenoma and benign phyllodes tumor are kinds of fibroepithelial tumor which have overlapping histopathological features. Recently, core biopsy is commonly performed to determine breast tumor, including fibroepithelial tumor. Small amount of tissue and overlapped histopathological features often complicate the Pathologist in diagnosing both. This study aims to describe the histopathological appearance which needed to diagnose fibroadenoma and benign phyllodes tumor and to verify if the diagnosis of fibroadenoma and benign phyllodes tumor in core biopsy using scoring system is more accurate than without scoring system.Method: This study was a diagnostic test, in which 57 cases of fibroadenoma and benign phyllodes tumor which had undergone core biopsy and mastectomy/excision were re-assessed using and without using scoring system. Histopathologic features which assessed using scoring system were stromal cellularity, nuclear atypia, tissue fragmentation, fat infiltration, mitotic figure, stromal heterogeneity. Analytical statistic, diagnostic test, accuracy test and Kappa test were done.Results: Stromal cellularity, stromal heterogeneity and tissue fragmentation mostly found in benign phyllodes tumor and significantly different (p=0,001; p=0,000; p=0,021).There were significant differences between stromal cellularity (p=0,001), stromal heterogeneity (p=0,000), and tissue fragmentation (p=0,021) in diagnosis of benign phyllodes tumor. Specificity in scoring system increased by17,9 %. Positive predictive value, negative predictive value and accuracy increased in scoring system (11,9% and 5,1%). Area under curve (AUC) increased by 8,9%. Cohen's Kappa test between core biopsy diagnosis without using and using scoring system had low result(0,545). Conclusion: The increasing of specificity, positive predictive value, accuracy and AUC proved that core biopsy with scoring system is more accurate than without scoring. This can be used as reference to diagnose fibroadenoma and benign phyllodes tumor.;Background: Fibroadenoma and benign phyllodes tumor are kinds of fibroepithelial tumor which have overlapping histopathological features. Recently, core biopsy is commonly performed to determine breast tumor, including fibroepithelial tumor. Small amount of tissue and overlapped histopathological features often complicate the Pathologist in diagnosing both. This study aims to describe the histopathological appearance which needed to diagnose fibroadenoma and benign phyllodes tumor and to verify if the diagnosis of fibroadenoma and benign phyllodes tumor in core biopsy using scoring system is more accurate than without scoring system.Method: This study was a diagnostic test, in which 57 cases of fibroadenoma and benign phyllodes tumor which had undergone core biopsy and mastectomy/excision were re-assessed using and without using scoring system. Histopathologic features which assessed using scoring system were stromal cellularity, nuclear atypia, tissue fragmentation, fat infiltration, mitotic figure, stromal heterogeneity. Analytical statistic, diagnostic test, accuracy test and Kappa test were done.Results: Stromal cellularity, stromal heterogeneity and tissue fragmentation mostly found in benign phyllodes tumor and significantly different (p=0,001; p=0,000; p=0,021).There were significant differences between stromal cellularity (p=0,001), stromal heterogeneity (p=0,000), and tissue fragmentation (p=0,021) in diagnosis of benign phyllodes tumor. Specificity in scoring system increased by17,9 %. Positive predictive value, negative predictive value and accuracy increased in scoring system (11,9% and 5,1%). Area under curve (AUC) increased by 8,9%. Cohen's Kappa test between core biopsy diagnosis without using and using scoring system had low result(0,545). Conclusion: The increasing of specificity, positive predictive value, accuracy and AUC proved that core biopsy with scoring system is more accurate than without scoring. This can be used as reference to diagnose fibroadenoma and benign phyllodes tumor.;Background: Fibroadenoma and benign phyllodes tumor are kinds of fibroepithelial tumor which have overlapping histopathological features. Recently, core biopsy is commonly performed to determine breast tumor, including fibroepithelial tumor. Small amount of tissue and overlapped histopathological features often complicate the Pathologist in diagnosing both. This study aims to describe the histopathological appearance which needed to diagnose fibroadenoma and benign phyllodes tumor and to verify if the diagnosis of fibroadenoma and benign phyllodes tumor in core biopsy using scoring system is more accurate than without scoring system.Method: This study was a diagnostic test, in which 57 cases of fibroadenoma and benign phyllodes tumor which had undergone core biopsy and mastectomy/excision were re-assessed using and without using scoring system. Histopathologic features which assessed using scoring system were stromal cellularity, nuclear atypia, tissue fragmentation, fat infiltration, mitotic figure, stromal heterogeneity. Analytical statistic, diagnostic test, accuracy test and Kappa test were done.Results: Stromal cellularity, stromal heterogeneity and tissue fragmentation mostly found in benign phyllodes tumor and significantly different (p=0,001; p=0,000; p=0,021).There were significant differences between stromal cellularity (p=0,001), stromal heterogeneity (p=0,000), and tissue fragmentation (p=0,021) in diagnosis of benign phyllodes tumor. Specificity in scoring system increased by17,9 %. Positive predictive value, negative predictive value and accuracy increased in scoring system (11,9% and 5,1%). Area under curve (AUC) increased by 8,9%. Cohen's Kappa test between core biopsy diagnosis without using and using scoring system had low result(0,545). Conclusion: The increasing of specificity, positive predictive value, accuracy and AUC proved that core biopsy with scoring system is more accurate than without scoring. This can be used as reference to diagnose fibroadenoma and benign phyllodes tumor."

"ABSTRAK Latar belakang: Sarkoma Ewing merupakan suatu small round cell tumor yang ditandai dengan fusi gen EWSR1/FLI1 pada 85 kasus. Diagnosis akurat diperlukan karena memiliki respon baik terhadap protokol kemoterapi spesifik. Baku emas diagnosis sarkoma Ewing adalah deteksi translokasi spesifik dengan RT-PCR atau FISH, namun pemeriksaan tersebut belum tersedia di institusi kami, sehingga dilakukan upaya lain untuk mempertajam diagnosis. Secara morfologi, sarkoma Ewing sering overlapping dengan small round cell tumor lainnya. Pulasan CD99 merupakan penanda yang sangat sensitif untuk mendiagnosis sarkoma Ewing, namun juga sering overlapping dengan small round cell tumor lainnya. Beberapa penelitian mengemukakan FLI1 dapat digunakan sebagai penanda diagnosis sarkoma Ewing. Tujuan penelitian ini adalah menilai ekspresi FLI1 untuk membantu menegakkan diagnosis sarkoma Ewing pada small round cell tumor yang memberikan hasil positif terhadap CD99, terutama pada kasus biopsi.Bahan dan cara: Penelitian ini menggunakan desain potong lintang. Sampel terdiri atas 36 kasus sarkoma Ewing dan 18 kasus small round cell tumor yang sudah dilakukan pulasan imunohistokimia CD99 di RSCM dari Januari 2011 sampai Mei 2018. Dilakukan pulasan FLI1 dan penilaian menggunakan H-score.Hasil: Titik potong H-score pada ekspresi FLI1 didapatkan 226.1 75 dengan sensitivitas 81.6 dan spesifisitas 94.4 . Ekspresi FLI1 tinggi didapatkan pada 31 kasus sarkoma Ewing, sedangkan pada 18 kasus small round cell tumor umumnya memiliki ekspresi FLI1 yang rendah ABSTRACT Background: Ewing 39;s sarcoma is a small round cell tumor characterized by EWSR1 / FLI1 gene fusion in 85 of cases. Accurate diagnosis is necessary because it has a good response to a specific chemotherapy protocol. The gold standard of Ewing 39;s sarcoma diagnosis is the detection of specific translocation with RT-PCR or FISH, but the examination is not yet available at our institution, so another attempt is made to sharpen the diagnosis. Morphologically, Ewing 39;s sarcoma is often overlapping with other small round cell tumor. CD99 is a very sensitive marker for diagnosing Ewing 39;s sarcoma, but also often overlapping with other small round cell tumors. Several studies have suggested that FLI1 can be used as a marker of Ewing rsquo;s sarcoma. The purpose of this study was to assess the FLI1 expression to help establish the diagnosis of Ewing rsquo;s sarcoma in small round cell tumors that gave CD99 positive results, especially in the biopsy cases. Materials and methods: This was a cross-sectional study with 36 cases of Ewing rsquo;s sarcoma and 18 cases of other small round cell tumor that had been performed CD99 immunohistochemistry at RSCM from January 2011 to May 2018. All cases stained by FLI1 antibody and evaluated using H-score. Results: The H-score cut-off point on FLI1 expression was obtained at 226.1 75 with 81.6 sensitivity and 94.4 specificity. The high FLI1 expression was obtained in 31 cases of Ewing rsquo;s sarcoma, while in 18 cases of small round cell tumor were generally had low expression of FLI1 p"

"Latar belakang: CTC sebagai bagian dari liquid biopsy berperan dalam melakukan monitoring pasien kanker payudara yang menjalani terapi. Adanya CTC menjadi pertanda resistensi terapi dan memengaruhi prognosis pasien. Penelitian ini bertujuan melihat adakah perubahan nilai CTC pada pasien kanker payudara stadium lokal lanjut atau lanjut yang mendapatkan kemoterapi serta melihat perubahan nilai CTC tersebut apakah dipengaruhi oleh usia, status menopause, subtipe, metastasis, dan grade.Metode: Didapatkan 30 sampel pasien kanker payudara stadium lokal lanjut atau lanjut yang akan mendapatkan kemoterapi berbasis Anthracycline dan Taxan. Pre kemoterapi pasien diambil darah perifer dan dilakukan pemeriksaan CTC menggunakan flowcytometry dengan antibodi EpCAM. Pasien lalu menjalani siklus kemoterapi hingga lengkap. Setelah itu pasien kembali diambil darah perifer dan diperiksa nilai CTC post kemoterapi.Hasil: Dari ke 30 sampel, didapatkan mean usia 47,93+7.30. Sebanyak 56,7 (n=17) belum menopause, 43,3% status tumor T3 dan T4, status kelenjar getah bening terbanyak adalah N0 dan N1 (43,3%). Hanya 2 pasien yang ditemukan ada metastasis. 56,7% pasien dengan grade 3, dan subtipe terbanyak adalah luminal B ( 63,4%, n=19). Terdapat 22 pasien (73,3%) dengan ER positif, 14 pasien (46,7%) dengan PR positif. Terdapat 11 pasien (36,7%) dengan Her2 positif dan 21 pasien (70%) dengan Ki67 high proliferation. Hasil CTC pre kemoterapi didapatkan nilai median 1460,50 sedangkan CTC post kemoterapi didapatkan nilai median 415,50 dilakukan uji Wilcoxon dan perbedaan bermakna dengan nilai p=0,002. Analisis multivariat regresi linier dihubungkan antara penurunan nilai CTC terhadap usia, status menopause, subtipe, metastasis, dan grading didapatkan status menopause berhubungan bermakna terhadap perubahan nilai CTC (p<0,05).Kesimpulan: CTC pada pasien kanker payudara stadium lokal lanjut dan lanjut setelah kemoterapi lebih rendah bermakna dibandingkan sebelum kemoterapi. Status menopause memiliki hubungan bermakna terhadap penurunan jumlah CTC setelah kemoterapi pada kanker payudara stadium lokal lanjut dan lanjut.......Background: As part of liquid biopsy, CTCs play a role in monitoring breast cancer patients undergoing therapy. The existence of CTCs is a sign of therapy resistance and affects patient prognosis. This study aims to examine whether there are changes in CTC values in patients with locally advanced or advanced breast cancer, who receive chemotherapy and are influenced by age, menopause status, subtype, metastasis, and grade.Method: Of the 30 samples of locally advanced or advanced breast cancer patients receiving Anthracycline and Taxan-based chemotherapy were obtained. Pre-chemotherapy, peripheral blood, was drawn and CTCs were examined using flow cytometry with EpCAM antibody. Patients then undergo a complete chemotherapy cycle. After that, the patients were again taken peripheral blood and examined for post-chemotherapy CTC values.Result: The study was conducted at Cipto Mangunkusumo Hospital, started from December 2022 to December 2023. Of the 30 samples with the mean age was 47,93+7,30. A total of 56,7 (n=17) were not menopause, 43,3% of tumor status with the T3 and T4, and the most common lymph node status with the N0 and N1 (43,3%). Only two patients were found to have metastasis. Then, 56,7% of patients had grade 3, and the most common subtype was luminal B (63,4%, n=19). There were 22 patients (73,3%) with ER positive, 14 patients (46,7%) with PR positive, 11 patients (36,7%) with Her2 positive, and 21 patients (70%) with Ki67 high proliferation. Pre-chemotherapy CTC results obtained a median value of 1460.50, Meanwhile, post-chemotherapy CTC obtained a median value of 415,50. Wilcoxon test was performed and the difference was significant with a value of p = 0,002. Multivariate linear regression analysis was correlated between the decrease in CTC values with age, menopause status, subtype, metastasis, and grading. The menopausal status has a significant association with decrease CTC values (p<0,05).Conclusion: CTC in locally advanced and advanced breast cancer patients after chemotherapy was significantly lower than before chemotherapy. menopause status has a significant association with decreased CTC values after chemotherapy in locally advanced and advanced breast cancer."

"Kanker kolorektal merupakan jenis kanker dengan jumlah penderita terbanyak ketiga di dunia. Metode deteksi kanker kolorektal yang banyak digunakan yakni kolonoskopi, namun metode tersebut bersifat invasif dan mahal. Salah satu metode deteksi yang dapat digunakan sebagai alternatif adalah melalui deteksi penanda molekuler adalah CK20 pada Circulating Tumor Cells (CTC) yang terdapat dalam darah perifer. Permasalahan muncul akibat rendahnya jumlah CTC dalam darah. Deteksi gen CK20 pada serum darah berpotensi menjadi metode deteksi kanker kolorektal karena serum darah mengandung cell-free nucleic acid (cfNA) yang dilepaskan oleh sel tumor. Tujuan dari penelitian ini adalah untuk mendeteksi gen CK20 pada CTC dan serum darah dari penderita kanker kolorektal. Isolat RNA diisolasi dari sampel CTC dan serum darah yang berasal dari empat penderita kanker kolorektal RSUPN Cipto Mangunkusumo kemudian diamplifikasi melalui Quantitave RT-PCR dan dianalisis melalui metode Melt Curve Analysis (MCA). Hasil menunjukkan bahwa gen CK20 memiliki Tm ~81°C dan terdeteksi pada satu sampel serum dan dua sampel jaringan. Berdasarkan hasil yang diperoleh, dapat disimpulkan bahwa gen CK20 tidak terdeteksi pada CTC dan terdeteksi pada satu serum darah penderita kanker kolorektal.......Colorectal cancer occupies the third-highest number of cancer patients worldwide. The most widely used colorectal cancer detection method is colonoscopy. However, colonoscopy is invasive and expensive. One alternative detection method is the detection of molecular markers such as CK20 in circulating tumor cells (CTC) found in the peripheral blood. The problem arises due to the low number of CTCs in the blood. Detection of the CK20 gene in blood serum is potentially used as a colorectal cancer detection method because blood serum contains cell-free nucleic acid (cfNA) released by tumor cells. This study aimed to detect the CK20 gene in CTCs and blood serum from colorectal cancer patients. RNA isolates were isolated from CTC samples and blood serum from four colorectal cancer patients at Cipto Mangunkusumo Hospital and then amplified by quantitative RT-PCR and then analyzed using the Melt Curve Analysis method. The results showed that the CK20 gene had a melting temperature of an average of ~81°C and was detected in one blood serum sample and two cancer tissue samples. Based on the results obtained, it can be concluded that the CK20 gene was undetected in CTCs and detected in one blood serum of colorectal cancer patients."

"Kanker kolorektal menempati urutan kedua dengan jumlah kasus tertinggi di dunia. Kanker kolorektal dapat dideteksi dengan menggunakan kolonoskopi, namun metode tersebut bersifat invasif. Oleh karena itu, penggunaan liquid biopsy dapat digunakan sebagai metode alternatif karena tergolong semi-invasif. Metode liquid biopsy yang digunakan adalah menggunakan darah dari pembuluh perifer. Di dalam darah terdapat beragam komponen penyusun, salah satunya adalah Circulating Tumor Cells (CTC). Circulating Tumor Cells dapat dikarakterisasi dengan menggunakan penanda spesifik sitokeratin 20 (CK20) karena pendaran ekspresinya yang tinggi pada kanker kolorektal. Proses isolasi CTC dari sampel darah dapat dilakukan dengan bervariasi metode, salah satunya dengan red blood cells lysis (eritrolisis). Penelitian ini bertujuan untuk mengisolasi CTC dengan penanda CK20 dari sampel darah perifer serta mengetahui karakteristik dan jumlah CTC yang diperoleh dari subjek kanker kolorektal di RSUPN Dr. Cipto Mangunkusumo. Pada penelitian ini sampel darah diambil dari lima subjek kanker kolorektal. Untuk mengoptimalisasi hasil isolat CTC yang nantinya diperoleh maka dilakukan spike-in experiment dengan penambahan sel HT-29 ke dalam donor sehat. Hasil penelitian ini menunjukkan bahwa CTC ditemukan di seluruh subjek dengan rata-rata perolehan 6 CTC tunggal dan 1 CTC cluster dari 2 mL darah. Dengan demikian, immunofluorescence dapat digunakan untuk mengarakterisasi tipe morfologinya, yaitu CTC tunggal dan CTC cluster.......Colorectal cancer ranks second with the highest number of cases in the world. Colorectal cancer can be detected using colonoscopy, but this method is invasive. Therefore, liquid biopsy can be used as an alternative method because it is classified as semi-invasive. The liquid biopsy method used is using blood from peripheral vessels. In the blood there are various constituent components, one of which is Circulating Tumor Cells (CTCs). Circulating Tumor Cells can be characterized using the specific marker cytokeratin 20 (CK20) due to its high expression in colorectal cancer. The process of isolating CTCs from blood samples can be carried out using various methods, one of which is red blood cells lysis (erythrolysis). This study aims to isolate CTCs with the CK20 marker from peripheral blood samples and determine the characteristics and number of CTCs obtained from colorectal cancer subjects at RSUPN Dr. Cipto Mangunkusumo. In this study, blood samples were taken from five colorectal cancer subjects. To optimize the CTCs isolate results that will be obtained, a spike-in experiment was carried out by adding HT-29 cells to healthy donors. The results of this study showed that CTCs were found in all subjects with an average of 6 single CTCs and 1 cluster CTCs from 2 mL of blood. Thus, immunofluorescence can be used to characterize the morphological types, namely single CTCs and cluster CTCs."

"Kanker kolorektal (KKR) adalah kanker yang menyerang bagian kolon dan rektum pada usus besar. Kasus KKR menempati posisis keempat di Indonesia. Pada umumnya, KKR didiagnosis menggunakan pemeriksaan standar dengan kolonoskopi, namun metode ini bersifat invasif dan menimbulkan rasa tidak nyaman bagi penderita KKR. Oleh karena itu, diperlukan alternatif yang minim invasif dan mudah dilakukan dengan cara mendeteksi keberadaan Circulating Tumor Cells (CTC) dalam darah. Kelimpahan CTC dalam darah sangat rendah, diperkirakan 1—10 CTC per 10 mL darah, sehingga perlu dilakukan isolasi CTC untuk memperkaya jumlah CTC. Metode yang dapat digunakan untuk mengisolasi CTC, yaitu metode sentrifugasi gradien densitas termodifikasi SepMateTM-15. Dalam mendeteksi CTC diperlukan marker yang memiliki spesifisitas dan sensitivitas baik, serta berasal dari jaringan tumor primer penderita KKR, seperti CK20. Penelitian ini bertujuan untuk mengisolasi CTC dengan menggunakan metode gradien densitas termodifikasi SepMateTM-15 dan karakterisasi CTC dengan penanda CK20 dari darah perifer KKR menggunakan Imunofluorosensi. Hasil isolasi CTC menunjukkan seluruh subjek KKR berhasil diisolasi dengan positivitas ≥1 CTC/ mL darah. Single CTC terdeteksi pada subjek 1,2, dan 4. Cluster CTC terdeteksi pada subjek 2,3, dan 5. Diameter rata-rata CTC yang diperoleh 10,82¼m. Sel CTC berhasil dikarakterisasi single CTC dan cluster CTC dengan penanda CK20 dalam analisis Imunofluorosensi.......Colorectal cancer (CRC) is a cancer that affects the colon and rectum in the large intestine. In Indonesia, CRC cases are ranked fourth. Generally, CRC is diagnosed using standard examinations colonoscopy, this procedure is invasive and uncomfortable for CRC patients. Therefore, an alternative that is minimally invasive and easy to perform is needed to detect the presence of Circulating Tumor Cells (CTCs).  The abundance of CTCs in blood is very low, estimated at 1—10 CTC/10 mL. Therefore, CTC isolation is required to increase the quantity of CTCs. A method to isolate CTCs is the SepMateTM-15 modified density gradient centrifugation method. To detect CTCs, markers with good specificity and sensitivity are needed originating from primary tumor tissues in CRC patients, such as CK20. This study aims to isolate CTCs using the SepMateTM-15 modified density gradient method and characterize CTCs with CK20 marker from peripheral blood using immunofluorescence. All CRC subjects were successfully isolated with positivity of  ≥1 CTC/mL. Single CTC in subjects 1, 2, and 4. CTC clusters were identified in subjects 2, 3, and 5. The average diameter of CTCs is 10.82 μm. Cell CTCs were successfully characterized between single CTCs and CTC clusters using a CK20 marker by immunofluorescence analysis."

"Latar Belakang: Kanker kolorektal (KKR) adalah salah satu penyebab utama morbiditas dan mortalitas terkait kanker. Pemeriksaan baku emas yang dilakukan untuk mendiagnosis kanker kolorektal hingga saat ini adalah histopatologi, yang didapat dari biopsi saat kolonoskopi. Oleh karena tidak semua pasien mau untuk menjalani pemeriksaan invasif di awal rencana diagnosis, maka pendekatan non-invasif baru yang dapat melengkapi dan meningkatkan strategi untuk diagnosis non invasif dan manajemen kanker kolorektal sangat dibutuhkan. Circulating tumor cell (CTC) dalam darah diharapkan dapat digunakan sebagai penanda diagnostik yang non-invasif pada pasien dengan KKR.Tujuan: Penelitian ini merupakan uji diagnostik CTC pada KKR menggunakan metode isolasi dan analisis CTC imunomagnetik seleksi negatif menggunakan Easysep™ dan marker mesenkimal kanker CD44.Metode: potong lintang dengan populasi terjangkau pasien dewasa yang diduga kanker kolorektal di RSCM selama bulan September 2020 hingga September 2021. Analisis uji diagnostik digunakan untuk mencari titik potong beserta nilai sensitivitas, spesifisitas, nilai duga positif (NDP), nilai duga negatif (NDN), rasio kemungkinan positif (RKP), dan rasio kemungkinan negatif (RKN) CTC dalam mendeteksi KKR.Hasil: Sebanyak total 80 subjek penelitian, didapatkan mean umur 56 ± 11 tahun. Proporsi subjek berdasarkan jenis kelamin yaitu 46,3% pasien perempuan dan 53,8% pasien lelaki. Sebanyak 77,5% subjek mengalami kanker kolorektal, 7,5% polip adenoma, dan 15% polip inflamasi/ hiperplastik. Uji diagnostik CTC untuk mendeteksi KKR (KKR dibandingkan dengan polip inflamasi/hiperplastik + polip adenoma), dengan titik potong CTC >1,5 sel/mL, didapatkan hasil sensitivitas, spesifisitas, NDP, NDN, RKP, RKN berturut-turut sebesar 50%; 88,89%; 93,94%; 34,04%; 4,5; dan 0,56. Selain itu didapatkan variabel diferensiasi kanker memiliki hubungan yang signifikan (p<0,05) dengan jumlah CTC.Simpulan: Pemeriksaan CTC memiliki sensitifitas yang rendah dan spesifisitas yang tinggi untuk mendiagnosis KKR. Pemeriksaan ini dapat dipertimbangkan menjadi pemeriksaan penunjang diagnostik non-invasif kanker kolorektal sebelum menjalani pemeriksaan yang lebih invasif yaitu kolonoskopi dan biopsi histopatologi.......Background: Colorectal cancer (CRC) is one of the leading causes of cancer-related morbidity and mortality. The gold standard for diagnosing colorectal cancer is histopathology, which is obtained from a biopsy during colonoscopy. Since not all patients are willing to undergo invasive testing at the outset of the first diagnotic plan, new non-invasive approaches that can complement and enhance strategies for non-invasive diagnosis and management of colorectal cancer are urgently needed. Circulating tumor cells (CTC) in the blood are expected to be used as a non-invasive diagnostic marker in patients with CRC.Purpose: This research is a diagnostic test of CTC on CRC using the isolation and analysis of negative selection immunomagnetic method with Easysep™ and CD44 cancer mesenchymal marker.Method: Cross-sectional study of adult patients with suspected colorectal cancer at RSCM during September 2020 to September 2021. Diagnostic study analysis was used to find the cut-off point along with sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), positive likelihood ratio (PLR), and negative likelihood ratio (NLR) of CTC in detecting CRC.Results: A total of 80 research subjects, the mean age was 56 ± 11 years. The proportion of subjects by gender was 46.3% female and 53.8% male patients. A total of 77.5% of the subjects had colorectal cancer, 7.5% adenoma polyps, and 15% inflammatory/ hyperplastic polyps. The CTC diagnostic analysis to detect CRC (CRC compared with inflammatory/hyperplastic polyps + adenoma polyps), with a CTC cut-off point of  > 1.5 cells/mL, showed that the sensitivity, specificity, PPV, NPV, PLR, NLR were 50%; 88.89%; 93.94%; 34.04%; 4.5; and 0.56, respectively. In addition, it was found that the cancer differentiation variable had a significant relationship (p<0.05) with the number of CTCs.Conclusion: CTC examination has low sensitivity and high specificity for diagnosing CRC. This examination can be considered as a non-invasive diagnostic support for colorectal cancer before undergoing more invasive examinations, which are colonoscopy and histopathological biopsy."