Identification and Validation of Reference Genes Selection in Ovarian Cancer Exposed to Hypoxia

Overview

Journal Onco Targets Ther

Publisher Dove Medical Press

Specialty Oncology

Date 2020 Aug 18

PMID 32801765

Citations 3

Authors

Wenying Yan

Mei Xie

Rong Li

Hongmei Hu

Biao Tang

Jie Shen

Affiliations

Soon will be listed here.

Abstract

Introduction: Hypoxia-mediated tumor metastasis, progression and drug resistance are major clinical challenges in ovarian cancer. Meanwhile, the genetic basis of these traits is still not clear. RT-qPCR, as an efficient and sensitive gene expression technique, has been widely used for gene analyses, providing a basis for in-depth understanding of molecular changes in different microenvironments. However, there is currently a lack of suitable reference genes to normalize the data associated with hypoxia in ovarian cancer cells.

Methods: A systematic method is needed to select the most suitable reference gene. Here, eight candidate reference genes (GAPDH, β-actin, 18S RNA, TUBB, PPIA, TBP, RPL13A and SDHA) from humans were selected to assess their expression levels in SKOV3 cells under hypoxia. The geNorm and NormFinder programs were utilized to evaluate the expression stabilities of these selected candidate reference genes.

Results: Interestingly, 18S RNA was considered to be an ideal reference gene for the normalization of target gene expression under hypoxic conditions. Furthermore, this result was confirmed in another two ovarian cancer cell line, CAOV3 and OVCAR3 cell line. Finally, these results suggest that appropriate reference genes should be selected before performing gene expression analysis during hypoxic environmental exposure.

Conclusion: 18S RNA can be used as an appropriate reference gene for the study of gene expression in ovarian cancer samples under hypoxia by RT-qPCR.

Citing Articles

Selection of Reference Genes for Normalization of Gene Expression After Exposure of Human Endothelial and Epithelial Cells to Hypoxia.

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Single-cell and transcriptomic analyses reveal the influence of diabetes on ovarian cancer.

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Uncovering the stability status of the reputed reference genes in breast and hepatic cancer cell lines.

Gorji-Bahri G, Moradtabrizi N, Hashemi A PLoS One. 2021; 16(11):e0259669.

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