Abstract：
:BACKGROUND: We describe an alternative method to determine mRNA half-life (t1/2) based on the Real-Time RT-PCR procedure. This approach was evaluated by using the beta-actin gene as a reference molecule for measuring of mRNA stability. RESULTS: Human leukemia Nalm-6 and CCRF-CEM cells were treated with various concentrations of Actinomycin D to block transcription and aliquots were removed periodically. Total RNA was isolated and quantified using the RiboGree&ncircledR; fluorescent dye with the VersaFluor Fluorometer System. One &mgr;g of total RNA was reverse transcribed and used as template for the amplification of a region of the beta-actin gene (231 bp). To generate the standard curve, serial ten-fold dilutions of the pBactin-231 vector containing the cDNA amplified fragment were employed, beta-actin mRNAs were quantified by Real-Time RT-PCR using the SYB&RcircledR; Green I fluorogenic dye and data analyzed using the iCycle iQ system software. Using this method, the beta-actin mRNA exhibited a half-life of 6.6 h and 13.5 h in Nalm-6 and CCRF-CEM cells, respectively. The t1/2 value obtained for Nalm-6 is comparable to those estimated from Northern blot studies, using normal human leukocytes (5.5 h). CONCLUSIONS: We have developed a rapid, sensitive, and reliable method based on Real-Time RT-PCR for measuring mRNA half-life. Our results confirm that beta-actin mRNA half-life can be affected by the cellular growth rate.
journal_name
Cancer Cell Intjournal_title
Cancer cell internationalauthors
Leclerc GJ,Leclerc GM,Barredo JCdoi
10.1186/1475-2867-2-1keywords:
subject
Has Abstractpub_date
2002-03-07 00:00:00pages
1issue
1issn
1475-2867journal_volume
2pub_type
杂志文章abstract：Background:MicroRNAs (miRNAs) play key roles in tumorigenesis and progression of gastric cancer (GC). miR-1269 has been reported to be upregulated in several cancers and plays a crucial role in carcinogenesis and cancer progression. However, the biological function of miR-1269 in human GC and its mechanism remain uncle...
journal_title：Cancer cell international
pub_type： 杂志文章
doi：10.1186/s12935-019-1026-4
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journal_title：Cancer cell international
pub_type： 杂志文章,评审
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journal_title：Cancer cell international
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journal_title：Cancer cell international
pub_type： 杂志文章
doi：10.1186/s12935-016-0376-4
更新日期：2017-01-05 00:00:00
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journal_title：Cancer cell international
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journal_title：Cancer cell international
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journal_title：Cancer cell international
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abstract：:[This corrects the article DOI: 10.1186/s12935-019-0742-0.]. ...
journal_title：Cancer cell international
pub_type： 杂志文章,已发布勘误
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更新日期：2019-04-11 00:00:00
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journal_title：Cancer cell international
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journal_title：Cancer cell international
pub_type： 杂志文章
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journal_title：Cancer cell international
pub_type： 杂志文章
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journal_title：Cancer cell international
pub_type： 杂志文章
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journal_title：Cancer cell international
pub_type： 杂志文章
doi：10.1186/1475-2867-6-16
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abstract：Background:The crucial role of long non-coding RNAs (lncRNAs) has been certified in human cancers. The lncRNAs with abnormal expressions could act as tumor inhibitors or oncogenes in the advancement of tumors. LBX2-AS1 was once reported to accelerate esophageal squamous cell carcinoma. Nonetheless, its function in gast...
journal_title：Cancer cell international
pub_type： 杂志文章
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journal_title：Cancer cell international
pub_type： 杂志文章
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journal_title：Cancer cell international
pub_type： 杂志文章
doi：10.1186/s12935-020-01268-x
更新日期：2020-05-24 00:00:00
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journal_title：Cancer cell international
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journal_title：Cancer cell international
pub_type： 杂志文章
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journal_title：Cancer cell international
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journal_title：Cancer cell international
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pub_type： 杂志文章
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更新日期：2018-12-22 00:00:00
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journal_title：Cancer cell international
pub_type： 杂志文章
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更新日期：2016-03-10 00:00:00
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journal_title：Cancer cell international
pub_type： 杂志文章
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journal_title：Cancer cell international
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更新日期：2014-11-30 00:00:00
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journal_title：Cancer cell international
pub_type： 杂志文章
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journal_title：Cancer cell international
pub_type： 杂志文章
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更新日期：2015-05-23 00:00:00
abstract：Background:Angiogenesis plays critical roles in the progression and metastasis of malignant tumors. Gastric neuroendocrine carcinoma is an uncommon stomach cancer that is rich in blood vessels and exhibits highly malignant biological behavior with a poor prognosis. The role of CDK5RAP3 in GNEC has not been reported to ...
journal_title：Cancer cell international
pub_type： 杂志文章
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