Tesis

Perbedaan Keragaman dan Komposisi Mikrobiota Mukosa Kolon Pasien Kanker Kolorektal dan Non Kanker Kolorektal = Differences in Diversity and Composition of MucosaAssociated Colonic Microbiota in Colorectal Cancer and Non-Colorectal Cancer.

Latar Belakang Kanker kolorektal merupakan keganasan ketiga paling sering ditemukan dan keganasan keempat paling sering menyebabkan kematian di dunia. Beberapa penelitian menunjukkan hubungan mikrobiota usus dengan kanker kolorektal. Hasil penelitian yang ada masih berbeda-beda oleh karena mikrobiota dipengaruhi beberapa faktor seperti kebiasan, pola makanan termasuk faktor geografis. Oleh karena itu penting mengenali perbedaan keragaman dan komposisi mikrobiota local yang nantinya dapat digunakan dalam diagnosis, pencegahan maupun pertimbangan pemilihan terapi. Tujuan Mengetahui perbedaan keragaman dan komposisi mikrobiota mukosa kolon pada pasien dengan dan tanpa kanker kolorektal. Metode Penelitian menggunakan desain potong lintang, peneliti melakukan pengambilan sampel secara konsekutif pada 59 subyek (35 kanker kolorektal, dan 24 non-kanker kolorektal) pada pasien yang terindikasi kolonoskopi di Pusat Endoskopi saluran cerna RSCM dan RS Fatmawati. Pemeriksaan mikrobiota menggunakan 16s rRNA dan analisis bioinformatika menggunakan pipeline wf-metagenomics dari EPI2Me-Labs (Oxford Nanopore Technologies). Keragaman alfa berdasarkan Indeks Shannon dan Simpson, serta keragaman beta menggunakan PCoA. Hasil Dari 59 pasien, median usia 55 (18-79) tahun dengan jenis kelamin laki-laki ditemukan pada 54,2% dari keseluruhan subjek penelitian. Berdasarkan indeks Shannon, (3,28 vs 2,82, p > 0.05) dan indeks Simpson (0,050 vs 0,060, p > 0,05) pasien kanker kolorektal memiliki keragaman lebih tinggi mesti tidak bermakna secara statistik. Berdasarkan Analisa PCoA, peneliti menemukan perbedaan keragaman beta pada tingkat genus dan spesies dengan nilai permanova p < 0.05. Dari keseluruhan subjek penelitian peneliti menemukan 4 filum terbanyak yang mendominasi yaitu Firmicutes, Proteobacteria, Bacteriodota dan Fusobacteria namun tidak tampak perbedaan bermakna antara kedua kelompok. Genus Bacteriodes, Campylobacter, Peptostreptococcus, dan Parvimonas ditemukan lebih banyak pada pasien dengan kanker kolorektal (p < 0.05), sedangkan Faecalibacterium, Haemophilus, dan Phocaeicola lebih banyak pada pasien non kanker kolorektal. (p < 0.05). Relative abundance spesies Fusobacterium nucleatum, Bacteriodes fragilis, Enterococcus faecalis, dan Campylobacter hominis lebih tinggi secara bermakna pada pasien dengan kanker kolorektal. Sedangkan Faecalibacterium prausnitzii, dan Prevotella copri lebih banyak pada pasien tanpa kanker kolorektal. P < 0.05. Kesimpulan Terdapat peningkatan kelimpahan genus Bacteriodes, Campylobacter, Peptostreptococcus, dan Parvimonas, spesies Fusobacterium nucleatum, Bacteriodes fragilis, Enterococcus faecalis, dan Campylobacter hominis pada kanker kolorektal. Terdapat peningkatan genus Faecalibacterium, Haemophilus, Phocaeicola, Faecalibacterium prausnitzii, dan Prevotella copri pada pasien non-kanker kolorektal
Kata kunci: Mikrobiota mukosa kolon, keragaman, Kelimpahan


Backgrounds Colorectal cancer is the third most common malignancy and the fourth leading cause of death in the world. Several studies showed an association between gut microbiota and colorectal cancer. The gut microbiota is unique and distinctive and is influenced by geographic factors and habits. The results of existing research still vary because the microbiota is influenced by several factors such as habits, food patterns, including geographical factors. Therefore, it is important to recognize differences in the diversity and composition of local microbiota which can later be used in diagnosis, prevention and consideration of therapy selection. Objectives The aim of this study is to determine the differences in diversity and composition of the colonic mucosal microbiota in patients with and without colorectal cancer. Methods This study was a cross-sectional in which sampling was carried out consecutively on 59 subjects (35 colorectal cancer and 24 non-colorectal cancer in patients who were recommended for colonoscopy at RSCM Gastrointestinal Endoscopy Center and Fatmawati Hospital. Microbiota examination was carried out using 16s rRNA. Bioinformatics analysis was performed using the wf-metagenomics pipeline from EPI2MeLabs (Oxford Nanopore Technologies). Alpha diversity was assessed using Shannon and Simpson indices, and beta diversity was evaluated using PCoA. Results Of the 59 patients, the median age was 55 (18-79) years-old with 54.2% subject was male. Based on the Shannon index, colorectal cancer patients had a greater median index value than non-colorectal patients (3.28 vs 2.82, p > 0.05) whereas based on the Simpson index, colorectal patients had a smaller value (0.050 vs 0.060, p > 0.05). PCoA analysis showed differences in beta diversity at genus and species levels with a permanova value p < 0.05. In this study, 4 phyla were found to dominate among all subjects, namely Firmicutes, Proteobacteria, Bacteriodota and Fusobacteria, but there were no significant differences between the two groups. The genera Bacteriodes, Campylobacter, Peptostreptococcus, and Parvimonas were found more frequently in patients with colorectal cancer (p < 0.05), while Faecalibacterium, Haemophilus, and Phocaeicola were more frequently found in noncolorectal cancer patients. (p < 0.05). The relative abundance of Fusobacterium nucleatum, Bacteriodes fragilis, Enterococcus faecalis, and Campylobacter hominis species were significantly higher in patients with colorectal cancer. Meanwhile, Faecalibacterium prausnitzii and Prevotella copri were more commonly found in non-colorectal cancer patients. P < 0.05. Conclusion There was an increase in the abundance of the Bacteriodes, Campylobacter, Peptostreptococcus, and Parvimonas, Fusobacterium nucleatum, Bacteriodes fragilis, Enterococcus faecalis, and Campylobacter hominis in colorectal cancer patients, while in non-colorectal cancer patients, there was an increase in the Faecalibacterium, Haemophilus, Phocaeicola, Faecalibacterium prausnitzii, and Prevotella copri.
Key words: Colonic mucosal microbiota, diversity, abundanc