Application of miR-34c in preparation of medicaments or health-care products for treating or preventing liver cancers
1. The technology of mir-34c and health products, applied in the application field of primary liver cancer, can solve the problems that there is no research on the regulation mechanism of miR-34 liver cancer genes, and it is difficult to determine
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Embodiment 1
[0019] Example 1: Gene chip detection of expression of miRNAs and mRNAs in HBV transgenic Balb / c mouse liver cancer tissue and normal mouse liver tissue
[0020] According to the instructions of Trizol reagent, extract the total RNA of cells: take 6, 12, and 20-month-old HBV transgenic Balb / c mouse liver tissue and normal Balb / c mouse liver tissue of bean size, add 1ml Trizol extract, grind, Pipette and mix well, collect the suspension into DEPC-treated Ep tubes, and incubate the samples at 15°C to 30°C for 5 minutes to fully separate the nucleic acid and protein. Add 0.2ml chloroform, shake vigorously for 15s, incubate at 15°C-30°C for 2-3min, centrifuge at 12000rpm at 4°C for 15min, take the RNA supernatant, transfer it to a new Ep tube, add 0.5ml isopropanol, mix upside down Evenly, incubate at 15°C-30°C for 10min, centrifuge at 12000rpm at 4°C for 10min, discard the supernatant, add 1ml 75% DEPC-ethanol, rinse the precipitate, centrifuge at 7500rpm, 2°C-8°C for 5min. Disc...
Embodiment 2
[0022] Example 2: Prediction of miR-34c target genes
[0023] In conjunction with the result of embodiment 1 and bioinformatics method, utilize online database (TargetScan: http: / / www.targetscan.org / ; Pictar: http: / / pictar.bio.nyu.edu / ; miRanda: http: / / microrna.sanger.ac.uk / ) predicts the target gene related to miR-34c, tentatively confirms that the target gene of miR-34c is TGIF 2 .
Embodiment 3
[0024]Example 3: Real-time quantitative RT-PCR detection of miR-34c and TGIF in liver cancer cell lines 2 The expression level
[0025] The total RNA of liver cancer cell lines HepG2 and HepG2.2.15 cells were extracted by Trizol method. Wash the cells twice with PBS, add 1ml Trizol to each bottle, pipette to mix, and incubate at room temperature for 5min; transfer the cell solution into a DEPC-treated 1.5ml centrifuge tube, add 0.2ml chloroform to each tube, shake vigorously for 15s to mix, Stand on ice for 3-5 minutes to separate layers; centrifuge at 12,000 rpm for 15 minutes at 4°C; carefully absorb the upper layer liquid, transfer it to another DEPC-treated 1.5ml centrifuge tube, add an equal volume (0.5ml) that has been pre-prepared at 4°C Cold isopropanol, upside down and gently mix for 15-30s, let stand on ice (room temperature) for 5-10min; 4℃, 12000rpm, centrifuge for 15min; discard the supernatant, add 1ml 75% ethanol, gently wash the precipitate 2 centrifuge at 75...
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