Identification of Suitable Reference Genes for qPCR Analysis of Serum microRNA in Gastric Cancer Patients

Song, Jianning; Bai, Zhigang; Han, Wei; Zhang, Jun; Meng, Hua; Bi, Jintao; Ma, Xuemei; Han, Song; Zhang, Zhongtao

doi:10.1007/s10620-011-1981-7

Cited by 248 publications

(223 citation statements)

References 34 publications

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“…Based on the different normalization strategies, miR-101a was determined to be the most optimal and stable reference gene. Furthermore, mir-101a was also demonstrated to be ideal for normalization of serum microRNA in gastric cancer patients (38).…”

Section: A B Cmentioning

confidence: 99%

Identification of suitable reference genes for BDV-infected primary rat hippocampal neurons

Mao

Zhang

Guo

et al. 2016

Molecular Medicine Reports

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Abstract. Borna disease virus (BDV) is a neurotropic RNA virus that infects the limbic system of mammals and results in behavioral disorders. The hippocampus is a core region in the limbic system, which contributes to memory and learning and is important in the regulation of emotion. However, no validated microRNA housekeeping genes have yet been identified in BDV-infected rat primary hippocampal neurons. Proper normalization is key in accurate miRNA expression analysis. The present study used reverse transcription-quantitative polymerase chain reaction (RT-qPCR) to evaluate the expression stability of 10 commonly used reference genes 5S, U6, E2 small nucleolar RNA (snoRNA), in BDV-infected rat hippocampal neurons and non-infected controls across 12 days post-infection. The data was analyzed by four statistical algorithms: geNorm, NormFinder, BestKeeper, and the comparative Δ-Ct method. Subsequently, the most suitable reference genes (miR-101a and U87) and the least suitable (snoRNA) were determined by the RankAggreg package. miR-155 was selected as a standard by which to evaluate the most and least suitable reference genes. When normalized to the most stable reference gene there were significant differences between the two groups. However, when the data were normalized to the less stably expressed gene, the results were not significant. miR-101a was recommended as a suitable reference gene for BDV-infected rat primary hippocampal neurons.

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Section: A B Cmentioning

confidence: 99%

Identification of suitable reference genes for BDV-infected primary rat hippocampal neurons

Mao

Zhang

Guo

et al. 2016

Molecular Medicine Reports

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“…In their study targeting serum miRNA in gastric cancer patients, Song et al demonstrated that RNU6B could not be detected in almost half of the serum samples; in the other half of patients, it was detected with Ct values of >40. Moreover, RNU6B was less stably expressed than let-7a and miR-16 in normal and cancerous human solid tissues (25). Although we actually examined the same study using RNU6B for normalization, there was marked variation in Ct values when comparing CCC patients and HCs (Fig.…”

Section: Discussionmentioning

confidence: 98%

Serum miR-210 as a potential biomarker of early clear cell renal cell carcinoma

Iwamoto

Kanda

Sejima

et al. 2013

International Journal of Oncology

103

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Abstract. Early detection and treatment are critical in the management of renal cell carcinoma (RCC). However, there is no standard serum biomarker to facilitate early diagnosis or prognostic stratification in patients with RCC. Recent reports suggest that circulating microRNAs (miRNAs) have great potential as biomarkers for diagnosis and prognosis in patients with several types of cancers. Further, many studies using miRNA microarray analysis demonstrated that miR-210 expression in clear cell carcinoma (CCC), which is the largest subtype of RCC, was significantly upregulated in tumor tissue. Therefore, we investigated whether serum miR-210 could be a useful biomarker for the diagnosis and progression of CCC. This study included 34 CCC patients and 23 healthy controls (HC). First, we analyzed tissue miR-210 levels in tumor tissues and matched normal tissues from the 34 CCC patients. Second, we investigated the serum miR-210 levels in the 34 CCC patients and the 23 HC patients. Real-time polymerase chain reaction (PCR) was used to measure miRNA levels. Moreover, we examined the correlation between serum miR-210 levels and the clinicopathological parameters. Among patients with CCC, expression of miR-210 was higher in tumor tissues compared to normal tissues (P<0.001). Serum miR-210 levels were higher in CCC patients compared to HCs (P=0.001). Receiver operating characteristic (ROC) curve analysis showed an area under the ROC curve (AUC) of 0.77 (95% confidence interval, 0.65-0.89) and a sensitivity and specificity of 65 and 83%, respectively. In addition, there was no significant association between serum miR-210 levels and age, sex, tumor size or existence of metastasis at diagnosis among the 34 CCC patients. In conclusion, serum miR-210 upregulation may occur in the early stage of CCC and serum miR-210 can be a useful biomarker for early CCC in humans.

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“…miR-103 (5p, TaqManID: 121218) and miR-191 (5p, TaqManID: 002299) [10]. All selected miRNAs are identified as potential biomarkers for various diseases, confirming there relative abundancy in serum [9,[11][12][13][14].…”

Section: Patients and Mirna Selectionmentioning

confidence: 92%

Serum miRNAs as Potential Biomarkers for the Bronchiolitis Obliterans Syndrome after Lung Transplantation

Budding

Rossato

Graaf

et al. 2016

The Journal of Heart and Lung Transplantation

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A B S T R A C TLung transplantation (LTx) is the last treatment for patients suffering from end-stage lung diseases. Survival postLTx is hampered by the development of the bronchiolitis obliterans syndrome (BOS) and diagnosis is often late. Given the urgent clinical need to recognize BOS patients at an early stage, we analyzed circulating miRNAs to identify possible stratification markers for BOS development post-transplantation. Therefore, pro-fibrotic (miR-21, miR-155), anti-fibrotic (miR-29a) and fibrosis-unrelated (miR-103, miR-191) miRNAs were analyzed in serum of end-stage lung disease patients and during LTx follow-up.Significant elevated levels of serum miRNAs were observed for all investigated miRNAs in both chronic obstructive pulmonary disease and interstitial lung disease patients compared to healthy controls. The same miRNAs were also significantly increased in the serum of BOS + vs. BOS− patients. Most importantly, miR-21, miR-29a, miR-103, and miR-191 levels were significantly higher in BOS+ patients prior to clinical BOS diagnosis.We demonstrated that a selected group of miRNAs investigated is elevated in end-stage lung disease and BOS + patients, prior to clinical BOS diagnosis. Even if further research is expedient on the prognostic value of circulating miRNAs in BOS and lung conditions in general, these results strongly suggest that circulating miRNAs could be used as potential biomarkers for BOS development.

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Identification of Suitable Reference Genes for qPCR Analysis of Serum microRNA in Gastric Cancer Patients

Cited by 248 publications

References 34 publications

Identification of suitable reference genes for BDV-infected primary rat hippocampal neurons

Identification of suitable reference genes for BDV-infected primary rat hippocampal neurons

Serum miR-210 as a potential biomarker of early clear cell renal cell carcinoma

Serum miRNAs as Potential Biomarkers for the Bronchiolitis Obliterans Syndrome after Lung Transplantation

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