Isolation and Identification of Long Non-Coding RNAs in Exosomes Derived from the Serum of Colorectal Carcinoma Patients.

Chin Tat Ng,Wai Kien Yip,Shamin Azwar,Heng Fong Seow,Siti Yazmin Zahari Sham,Mohd Faisal Jabar,Norhafizah Mohtarrudin,Norren Haneezah Sahak

doi:10.3390/biology10090918

Chin Tat Ng, Wai Kien Yip + Show 6 more

Open Access

https://doi.org/10.3390/biology10090918

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Journal: Biology	Publication Date: Sep 15, 2021
Citations: 9	License type: CC BY 4.0

Affiliation: Universiti Putra Malaysia, Serdang Hospital

Abstract

Simple SummaryTreatment regimens for patients with advanced disease are limited and the mortality rate is high in these patients. A better understanding on pathogenesis and progression of cancer is critical for the development of new treatment strategies. In colorectal cancer (CRC), exosomes (secreted vesicles from cells) and long non-coding RNAs (lncRNAs) have been shown to play significant roles in disease development and progression. Long non-coding RNAs (lncRNAs) are present in the exosomes of serum and their profiles may potentially be useful as novel biomarkers for CRC patients and may provide a new insight in the pathogenesis and progression of CRC. Here, we compared the expression profiles of exosomal lncRNAs between non-cancer individuals and patients with colorectal carcinoma. The relative expression level of LINC00152 was found to be significantly lower in exosomes from sera of CRC patients as compared to non-cancer individuals whereas lncRNA H19 was significantly up-regulated in advanced-stages (stage III and IV) of CRC as compared to early-stages (stage I and II). Our data suggest that LINC00152 and H19 may play important roles in pathogenesis and progression of CRC.Long non-coding RNAs (lncRNAs) are non-coding RNAs consisting of more than 200 nucleotides in length. LncRNAs present in exosomes may play a critical role in the cellular processes involved in cancer pathogenesis and progression including proliferation, invasion, and migration of tumor cells. This paper aims to identify the differential expression of exosomal lncRNAs derived from the sera of non-cancer individuals and patients diagnosed with colorectal carcinoma. These differentially-expressed exosomal serum lncRNAs may provide an insight into the pathogenesis and progression of colorectal cancer (CRC). Serum exosomes and exosomes from SW480-7 cell culture supernatants were isolated and viewed by transmission electron microscope (TEM). The particle size distribution and protein markers of exosomes derived from SW480-7 were further analyzed using the Zetasizer Nano S instrument and western blotting technique. TEM showed that exosomes derived from serum and SW480-7 cells were round vesicles with sizes ranging from 50–200 nm. The exosomes derived from SW480-7 had an average diameter of 274.6 nm and contained the exosomal protein, ALIX/PDCD6IP. In our clinical studies, six lncRNAs, namely GAS5, H19, LINC00152, SNHG16, RMRP, and ZFAS1 were detected in the exosomes from sera of 18 CRC patients. Among these six lncRNAs, the expression level of LINC00152 was found to be significantly lower in CRC patients as compared to non-cancer individuals (p = 0.04) while lncRNA H19 was significantly up-regulated in advanced-stages (stage III and IV) of CRC (p = 0.04) as compared to early-stages (stage I and II). In conclusion, the detection of lower LINC00152 in exosomes of sera from CRC patients versus non-cancer individuals and H19 upregulation in advanced stages suggests that they may play important roles in pathogenesis and progression of CRC.

Highlights

Colorectal cancer (CRC) is the third most common cancer worldwide [1] that contributes to a high mortality rate, despite decades of research and improvement in clinical management [2]
Our results revealed that SNHG16 was down-regulated in exosomes from sera of colorectal cancer (CRC) patients but there is no difference between early- and advanced-stage CRC
Our study has shown that Long non-coding RNAs (lncRNAs), H19 was up-regulated in serumderived exosomes from advanced-stage CRC, while exosomal LINC00152 was downregulated in CRC as compared with non-cancer individuals

Summary

Introduction

Colorectal cancer (CRC) is the third most common cancer worldwide [1] that contributes to a high mortality rate, despite decades of research and improvement in clinical management [2]. Recent literature in the pathogenesis of CRC has mainly focused on the involvement of an increasing number of non-coding ribonucleic acids (ncRNAs) via next-generation sequencing and transcriptome analysis [4,5]. These ncRNAs encompass a variety of subclasses that include short regulatory ncRNAs such as miRNAs, siRNAs, and piRNAs, as well as the long non-coding RNAs (lncRNAs). LncRNAs are transcripts of more than 200 nucleotides [6] in length with no or limited protein-coding potential and have recently been found in significant amounts inside exosomes. Exosomes carrying biomarkers specific to the origin of cancer cells [15]

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