ArticlePloS one2021
Uncovering the stability status of the reputed reference genes in breast and hepatic cancer cell lines.
Article in PloS one, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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13 citing papers in PubMed, 16 citations in OpenAlex.
- Advances in algorithms for normalizer gene selection in qRT-PCR: implications for cancer biology and precision medicine.Frontiers in genetics · 2026Review
- Identification of a nuclear transcriptional module associated with ERα and E2F/FOXM1 signaling across PAM50 breast cancer subtypes.Frontiers in oncology · 2026Article
- Evaluation of normalisation strategies for qPCR data obtained from canine gastrointestinal tissues with different pathologies.Scientific reports · 2025Article
- Identification of an appropriate reference gene for normalization of qRT-PCR expression analyses in human breast cancer cell lines: application to L-arginine depletion studies.Journal of cancer research and clinical oncology · 2025Article
- Screening and validating the optimal panel of housekeeping genes for 4T1 breast carcinoma and metastasis studies in mice.Scientific reports · 2024Article
- Reference gene evaluation for normalization of gene expression studies with lymph tissue and node‑derived stromal cells of patients with oral squamous cell carcinoma.Oncology letters · 2024Article
- GC-MS analysis, molecular docking, and apoptotic-based cytotoxic effect of Caladium lindenii Madison extracts toward the HeLa cervical cancer cell line.Scientific reports · 2024Article
- Selection of reference genes in liproxstatin-1-treated K562 Leukemia cells via RT-qPCR and RNA sequencing.Molecular biology reports · 2024Article
- Tumor suppressor miR-218 directly targets epidermal growth factor receptor (EGFR) expression in triple-negative breast cancer, sensitizing cells to irradiation.Journal of cancer research and clinical oncology · 2023Article
- Article
- Inverse Impact of Cancer Drugs on Circular and Linear RNAs in Breast Cancer Cell Lines.Non-coding RNA · 2023Article
- Role of PAX2 in breast cancer verified by bioinformatics analysis andAnnals of translational medicine · 2023Article
- Article
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Accurate and reliable relative gene expression analysis via the Reverse Transcription-quantitative Real Time PCR (RT-qPCR) method strongly depends on employing several stable reference genes as normalizers. Utilization of the reference genes without analyzing their expression stability under each experimental condition causes RT-qPCR analysis error as well as false output. Similar to cancerous tissues, cancer cell lines also exhibit various gene expression profiles. It is crucial to recognize stable reference genes for well-known cancer cell lines to minimize RT-qPCR analysis error. In this study, we showed the expression level and investigated the expression stability of eight common reference genes that are ACTB, YWHAZ, HPRT1, RNA18S, TBP, GAPDH, UBC, and B2M, in two sets of cancerous cell lines. One set contains MCF7, SKBR3, and MDA-MB231 as breast cancer cell lines. Another set includes three hepatic cancer cell lines, including Huh7, HepG2, and PLC-PRF5. Three excel-based softwares comprising geNorm, BestKeeper, and NormFinder, and an online tool, namely RefFinder were used for stability analysis. Although all four algorithms did not show the same stability ranking of nominee genes, the overall results showed B2M and ACTB as the least stable reference genes for the studied breast cancer cell lines. While TBP had the lowest expression stability in the three hepatic cancer cell lines. Moreover, YWHAZ, UBC, and GAPDH showed the highest stability in breast cancer cell lines. Besides that, a panel of five nominees, including ACTB, HPRT1, UBC, YWHAZ, and B2M showed higher stability than others in hepatic cancer cell lines. We believe that our results would help researchers to find and to select the best combination of the reference genes for their own experiments involving the studied breast and hepatic cancer cell lines. To further analyze the reference genes stability for each experimental condition, we suggest researchers to consider the provided stability ranking emphasizing the unstable reference genes.
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