Identification of appropriate housekeeping genes for quantitative RT-PCR analysis in MDA-MB-231 and NCI-H460 human cancer cell lines under hypoxia and serum deprivation

doi:10.31083/j.jmcm.2018.03.001

Cited by 4 publications

(6 citation statements)

References 39 publications

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“…PPIA along with ACTB was found to be the most stable reference gene for basal type breast cancer cell lines in hypoxic and serum deprived conditions [36]. However, our analysis revealed that the expression of PPIA was moderately stable and ranked ninth in the combined analysis (Supplementary File S3).…”

Section: Discussionmentioning

confidence: 80%

“…In total, thirteen reference genes were selected by searching for relevant literature related to various breast cancer cell lines [26][27][28][29][30][31][32][33][34][35][36][37][38]. Twelve more genes were selected from the TCGA dataset analysis and were referred to as Novel Candidate Reference genes.…”

Section: Selection Of Reference Genes and Primer Designmentioning

confidence: 99%

“…Breast cancer various cell lines Literature ACTB [26,29,32,34], CCSER2 [35], GAPDH [34], HNRNPL [37], HSP90AB1 [31], PCBP1 [37], PGK1 [30], PPIA [36], PUM1 [28,33,35],…”

Section: Cell Line Source Selected Candidate Reference Genesmentioning

confidence: 99%

“…However, with the advent of technology, better techniques using RNA-seq data have been employed to identify stable reference genes. Several studies have previously identified novel reference genes and/or validated conventional reference genes for the study of breast cancer [26][27][28][29][30][31][32][33][34][35][36][37][38].…”

Section: Introductionmentioning

confidence: 99%

“…In the present study, we identified and validated novel reference genes that could be used to normalize qPCR data in SK-BR-3 breast cancer cell line. Additionally, we compared expression stability of newly identified genes with previously reported reference genes [26][27][28][29][30][31][32][33][34][35][36][37][38] to select suitable candidate reference genes. Our study reports for the first time to our knowledge, a comprehensive analysis, combining previous and novel candidates studied over multiple successive passages (p), in replicate cultures S1 and S2, and validated in various hypoxic conditions for SK-BR-3 cell line.…”

Section: Introductionmentioning

confidence: 99%

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Identification of Novel Endogenous Controls for qPCR Normalization in SK-BR-3 Breast Cancer Cell Line

Jain

Mitre

Nitisa

et al. 2021

Genes

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Normalization of gene expression using internal controls or reference genes (RGs) has been the method of choice for standardizing the technical variations in reverse transcription quantitative polymerase chain reactions (RT-qPCR). Conventionally, ACTB and GAPDH have been used as reference genes despite evidence from literature discouraging their use. Hence, in the present study we identified and investigated novel reference genes in SK-BR-3, an HER2-enriched breast cancer cell line. Transcriptomic data of 82 HER2-E breast cancer samples from TCGA database were analyzed to identify twelve novel genes with stable expression. Additionally, thirteen RGs from the literature were analyzed. The expression variations of the candidate genes were studied over five successive passages (p) in two parallel cultures S1 and S2 and in acute and chronic hypoxia using various algorithms. Finally, the most stable RGs were selected and validated for normalization of the expression of three genes of interest (GOIs) in normoxia and hypoxia. Our results indicate that HSP90AB1, DAD1, PFN1 and PUM1 can be used in any combination of three (triplets) for optimizing intra- and inter-assay gene expression differences in the SK-BR-3 cell line. Additionally, we discourage the use of conventional RGs (ACTB, GAPDH, RPL13A, RNA18S and RNA28S) as internal controls for RT-qPCR in SK-BR-3 cell line.

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Section: Discussionmentioning

confidence: 80%

Section: Selection Of Reference Genes and Primer Designmentioning

confidence: 99%

“…Breast cancer various cell lines Literature ACTB [26,29,32,34], CCSER2 [35], GAPDH [34], HNRNPL [37], HSP90AB1 [31], PCBP1 [37], PGK1 [30], PPIA [36], PUM1 [28,33,35],…”

Section: Cell Line Source Selected Candidate Reference Genesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Identification of Novel Endogenous Controls for qPCR Normalization in SK-BR-3 Breast Cancer Cell Line

Jain

Mitre

Nitisa

et al. 2021

Genes

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Chinese Herbal Formulas Miao-Yi-Ai-Tang Inhibits the Proliferation and Migration of Lung Cancer Cells through Targetingβ-Catenin/AXIN and Presents Synergistic Effect with Cisplatin Suppressing Lung Cancer

Zhang

Tan

et al. 2020

BioMed Research International

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Objective. Lung cancer is one of the major causes of cancer deaths worldwide, and the five-year survival still remains low despite the improvement of screening, prevention, and treatment methods. Chinese herbal medicines have been widely used for tumor prevention and treatment. Miao-Yi-Ai-Tang (Miao) is a novel herbal formulation and shows a potential anticancer effect. Materials and Methods. Human Small Cell Lung Cancer Cell was used for study in vitro. After treatments by Miao and Cisplatin (DDP), the invasion, migration, proliferation, and apoptosis of cells were detected by transwell, wound healing, CCK-8, and flow cytometry, respectively. The expression of β-catenin, AXIN, and c-myc was detected by qRT-PCR and immunohistochemistry staining. Western blotting was applied for measuring the protein expression of β-catenin. Subcutaneously transplanted tumor model of lung cancer NCI-H446 was established to investigate the influence of Miao on tumor growth. Results. We found that Miao could inhibit invasion, migration, and proliferation and promote apoptosis of human lung cancer cells. Meanwhile, Miao and DDP presented synergy regulating the proliferation and apoptosis of lung cancer cells. The percentage of lung cancer cells in S and G2 stages was increased markedly by Miao. Besides, the expression of c-myc, AXIN, and β-catenin was markedly inhibited by Miao. Tumor growth in vivo was markedly inhibited by Miao. Conclusions. Chinese herbal formulas Miao could suppress lung cancer through targeting the β-catenin/AXIN signaling pathway. Therefore, our findings may provide a novel strategy for the prevention and treatment of lung cancer.

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Exosomes isolated from the plasma of remote ischemic conditioning rats improved cardiac function and angiogenesis after myocardial infarction through targeting Hsp70

Chen

Huang

et al. 2020

Aging

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Identification of appropriate housekeeping genes for quantitative RT-PCR analysis in MDA-MB-231 and NCI-H460 human cancer cell lines under hypoxia and serum deprivation

Cited by 4 publications

References 39 publications

Identification of Novel Endogenous Controls for qPCR Normalization in SK-BR-3 Breast Cancer Cell Line

Identification of Novel Endogenous Controls for qPCR Normalization in SK-BR-3 Breast Cancer Cell Line

Chinese Herbal Formulas Miao-Yi-Ai-Tang Inhibits the Proliferation and Migration of Lung Cancer Cells through Targetingβ-Catenin/AXIN and Presents Synergistic Effect with Cisplatin Suppressing Lung Cancer

Exosomes isolated from the plasma of remote ischemic conditioning rats improved cardiac function and angiogenesis after myocardial infarction through targeting Hsp70

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