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Knockdown of long non-coding RNA ANRIL inhibits tumorigenesis in human gastric cancer cells via microRNA-99a-mediated down-regulation of BMI1

Long non-coding RNA antisense non-coding RNA in the INK4 locus (ANRIL) has been reported to promote tumorigenesis via regulating microRNA (miR)-99a in gastric cancer cells. However, the role of each component involved in it is still not well understood. This study aimed to verify the role of ANRIL i...

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Published in:Brazilian journal of medical and biological research 2018-01, Vol.51 (10), p.e6839
Main Authors: Liu, Pei, Zhang, Mingming, Niu, Qinghui, Zhang, Fengjuan, Yang, Yuling, Jiang, Xiangjun
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description Long non-coding RNA antisense non-coding RNA in the INK4 locus (ANRIL) has been reported to promote tumorigenesis via regulating microRNA (miR)-99a in gastric cancer cells. However, the role of each component involved in it is still not well understood. This study aimed to verify the role of ANRIL in gastric cancer as well as the underlying mechanisms. ANRIL levels in clinical gastric cancer tissues and cell lines were tested by qPCR. Effects of ANRIL silence on cell viability, migration and invasion, apoptosis, and miR-99a expression in MKN-45 and SGC-7901 cells were measured using CCK-8, Transwell assay, flow cytometry, and qPCR assays, respectively. Then, effects of miR-99a inhibition on ANRIL-silenced cells were evaluated. B-lymphoma Mo-MLV insertion region 1 (BMI1) expression, after abnormal expression of ANRIL and miR-99a, was determined. Finally, expression of key proteins in the apoptotic, Notch, and mTOR pathways was assessed. ANRIL level was elevated in gastric cancer tissues and cell lines. Knockdown of ANRIL suppressed cell viability, migration, and invasion, and increased apoptosis through up-regulating miR-99a. Furthermore, ANRIL silence down-regulated BMI1 via up-regulating miR-99a. BMI1 silence down-regulated Bcl-2 and key kinases in the Notch and mTOR pathways and up-regulated p16 and cleaved caspases. We verified the tumor suppressive effects of ANRIL knockdown in gastric cancer cells via crosstalk with miR-99a. Together, we provided a novel regulatory mechanism for ANRIL in gastric cancer, in which ANRIL silence down-regulated BMI1 via miR-99a, along with activation of the apoptotic pathway and inhibition of the Notch and mTOR pathways.
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However, the role of each component involved in it is still not well understood. This study aimed to verify the role of ANRIL in gastric cancer as well as the underlying mechanisms. ANRIL levels in clinical gastric cancer tissues and cell lines were tested by qPCR. Effects of ANRIL silence on cell viability, migration and invasion, apoptosis, and miR-99a expression in MKN-45 and SGC-7901 cells were measured using CCK-8, Transwell assay, flow cytometry, and qPCR assays, respectively. Then, effects of miR-99a inhibition on ANRIL-silenced cells were evaluated. B-lymphoma Mo-MLV insertion region 1 (BMI1) expression, after abnormal expression of ANRIL and miR-99a, was determined. Finally, expression of key proteins in the apoptotic, Notch, and mTOR pathways was assessed. ANRIL level was elevated in gastric cancer tissues and cell lines. Knockdown of ANRIL suppressed cell viability, migration, and invasion, and increased apoptosis through up-regulating miR-99a. 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Furthermore, ANRIL silence down-regulated BMI1 via up-regulating miR-99a. BMI1 silence down-regulated Bcl-2 and key kinases in the Notch and mTOR pathways and up-regulated p16 and cleaved caspases. We verified the tumor suppressive effects of ANRIL knockdown in gastric cancer cells via crosstalk with miR-99a. Together, we provided a novel regulatory mechanism for ANRIL in gastric cancer, in which ANRIL silence down-regulated BMI1 via miR-99a, along with activation of the apoptotic pathway and inhibition of the Notch and mTOR pathways.</abstract><cop>Brazil</cop><pub>Associacao Brasileira de Divulgacao Cientifica (ABDC)</pub><pmid>30156609</pmid><doi>10.1590/1414-431X20186839</doi><oa>free_for_read</oa></addata></record>
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ispartof Brazilian journal of medical and biological research, 2018-01, Vol.51 (10), p.e6839
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subjects Antisense RNA
Apoptosis
Apoptosis - genetics
Apoptotic pathway
Bcl-2 protein
BIOLOGY
BMI1
Carcinogenesis - genetics
Cell Line, Tumor
Cell migration
Cholecystokinin
Cyclin-dependent kinase inhibitors
Development and progression
Down-Regulation
Flow cytometry
Gastric cancer
Gene Expression Regulation, Neoplastic
Genetic aspects
Humans
INK4 protein
Kinases
lncRNA ANRIL
Lymphocytes B
Lymphoma
MEDICINE, RESEARCH & EXPERIMENTAL
MicroRNA
MicroRNAs
MicroRNAs - metabolism
miR-99a
miRNA
Neoplasm Invasiveness
Non-coding RNA
Notch/mTOR pathway
Physiological aspects
RNA, Long Noncoding - genetics
Stomach cancer
Stomach Neoplasms - genetics
Stomach Neoplasms - metabolism
Stomach Neoplasms - pathology
TOR protein
TOR Serine-Threonine Kinases - metabolism
Transfection
Tumor cell lines
Tumorigenesis
Up-Regulation
title Knockdown of long non-coding RNA ANRIL inhibits tumorigenesis in human gastric cancer cells via microRNA-99a-mediated down-regulation of BMI1
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