AI Article Synopsis
- A new method using stem-loop reverse transcriptional real-time PCR (RT-PCR) was developed to quickly and accurately measure let-7a miRNA levels in gastric cancer.
- The study involved designing specific primers and probes for the PCR process, and it found that this method could detect very low amounts (as little as 10 copies) of let-7a miRNA in tiny samples of RNA.
- Results showed that let-7a miRNA levels were significantly reduced in gastric tumor tissues compared to normal tissues, suggesting the miRNA could be important in gastric cancer development.
Article Abstract
Aim: To establish an accurate and rapid stem-loop reverse transcriptional real-time PCR (RT-PCR) method to quantify human let-7a miRNA in gastric cancer.
Methods: According to the sequence of let-7a miRNA, the stem-loop reverse transcriptional primer, the primers and quantitative MGB probes of real-time PCR were designed and synthesized. The dynamic range and the sensitivity of quantitative reverse transcriptional real-time PCR were determined. The levels of let-7a miRNA were examined in 32 gastric carcinoma samples by stem-loop RT-PCR method.
Results: The dynamic range and sensitivity of the let-7a miRNA quantification scheme were evaluated, the result showed the assay could precisely detect 10 copies of mature let-7a miRNA in as few as 0.05 ng of total RNA of gastric mucosa. The results of specificity analysis showed no fluorescence signal occurred even though 50 ng of human genomic DNA was added to the reverse transcription (RT) reaction. The expression level of let-7a miRNA in gastric tumor tissues was significantly lower compared to normal tissues in 14 samples from 32 patients.
Conclusion: The stem-loop RT-PCR is a reliable method to detect let-7a miRNA which may play an important role in the development of gastric carcinoma.
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Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4395645 | PMC |
| http://dx.doi.org/10.3748/wjg.v13.i20.2883 | DOI Listing |
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