Real-time Monitoring of MiRNA Function in Pancreatic Cell Lines Using Recombinant AAV-based MiRNA Asensors

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2013 Jun 19

PMID 23776656

Citations 8

Authors

Jing Chen

Xinjuan Liu

Xue Chen

Zihao Guo

Juan Liu

Jianyu Hao

Jie Zhang

Affiliations

Soon will be listed here.

Abstract

Background: MicroRNAs (miRNAs) are reportedly involved in pancreatic ductal adenocarcinoma (PDAC) development. Current methods do not allow us to reliably monitor miRNA function. Asensors are adeno-associated virus (AAV) vector miRNA sensors for real-time consecutive functional monitoring of miRNA profiling in living cells.

Methods: miR-200a, -200b, -21, -96, -146a, -10a, -155, and -221 in three PDAC cell lines (BxPC-3, CFPAC-1, SW1990), pancreatic epithelioid carcinoma cells (PANC-1), and human pancreatic nestin-expressing cells (hTERT-HPNE) were monitored by Asensors. Subsequently, the real-time consecutive functional profile of all miRNAs was evaluated.

Results: Selected miRNAs were detectable in all cell lines with high sensitivity and reproducibility. In the three PDAC cell lines, BxPC-3, CFPAC-1, and SW1990, the calibrated signal unit of the eight miRNAs Asensors was significantly lower than that of the Asensor control. However, in PANC-1 cells, miR-200a and -155 showed upregulation of target gene expression at 24 hours after infection with the sensors; at 48 hours, miR-200b and -155 displayed upregulation of reporter expression; and at 72 hours, reporter gene expression was upregulated by miR-200a and -200b. The result that miRNA could upregulate gene expression was further confirmed in miR-155 of hTERT-HPNE cells. Furthermore, miRNA activity varied among cell/tissue types and time.

Conclusion: It is possible that miRNA participates in the pathophysiology of pancreatic cancer, but the current popular methods do not accurately reveal the real-time miRNA function. Thus, this report provided a convenient, accurate, and sensitive approach to miRNA research.

Citing Articles

MicroRNAs: emerging biomarkers and therapeutic targets in pancreatic cancer.

Yuan J, Yan K, Guo Y, Li Y Front Mol Biosci. 2024; 11:1457875.

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MicroRNA-155 and its exosomal form: Small pieces in the gastrointestinal cancers puzzle.

Guo J, Zhong L, Momeni M Cell Biol Toxicol. 2024; 40(1):77.

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MicroRNAs: Small but Key Players in Viral Infections and Immune Responses to Viral Pathogens.

Bauer A, Majumdar N, Williams F, Rajput S, Pokhrel L, Cook P Biology (Basel). 2023; 12(10).

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Adding recombinant AAVs to the cancer therapeutics mix.

Mulcrone P, Herzog R, Xiao W Mol Ther Oncolytics. 2022; 27:73-88.

PMID: 36321134 PMC: 9588955. DOI: 10.1016/j.omto.2022.09.009.

[Predictive value of miRNA-29a and miRNA-10a-5p for 28-day mortality in patients with sepsis-induced acute kidney injury].

Huo R, Dai M, Fan Y, Zhou J, Li L, Zu J Nan Fang Yi Ke Da Xue Xue Bao. 2017; 37(5):646-651.

PMID: 28539288 PMC: 6780479.

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