Application of micro RNA (Ribonucleic Acid) gene miR-1291 on predication of prognosis of breast cancer
A breast cancer and gene technology, applied in the field of biomarkers for personalized treatment, can solve problems such as different curative effects and toxic and side effects, prognosis, different curative effect characteristics, and failure to meet clinical requirements, etc., to achieve the effect of improving prognosis
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Embodiment 1
[0029] Embodiment 1, breast cancer prognosis prediction
[0030] Using mirVana RNA Isolation Kit (Applied Biosystem p / n AM1556), according to the operation steps provided in the kit, total RNA was extracted from surgical resection samples of breast cancer patients, and stored in aliquots.
[0031] RNA quality detection. UV absorbance assay was used to measure the absorbance at wavelengths of 260nm and 280nm to determine the concentration of RNA samples, and combined with agarose gel electrophoresis to detect the quality of RNA.
[0032] The chip hybridization experiment was carried out in strict accordance with the operation manual of Agilent Human 8*60K miRNA chip. The basic process includes dephosphorylation, sample denaturation, ligation labeling, chip hybridization, chip washing, chip scanning and other steps.
[0033] The chip results were scanned with an Agilent scanner, and the Feature Extraction software read the data Scanresolution 5 μm, PMT 100%, and processed and a...
Embodiment 2
[0035] Example 2, real-time quantitative PCR detection of mir-1291 gene expression
[0036] The extraction and quality detection of RNA were as described in Example 1. Using Ribo's mir-1291Bulge-Loop TM miRNA qPCR detection kit detection. Wherein, the cDNA chain is synthesized by reverse transcription using the stem convex loop primer, and the real-time quantitative PCR amplification is performed using the synthesized cDNA as a template. Each sample was tested three times, and the content of U6snRNA or 5S rRNA in each sample was detected as an internal reference to correct the loading error, analyze the relative expression level, and compare the expression differences of different patients. The results showed that in a group of 52 breast cancer patients, the difference in the expression of mir-1291 was 2.8 times between breast cancer patients without metastases within 5 years and those with metastases within 5 years (p value < 0.0001).
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