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Gene HRNBP3 and new application of expression product of gene HRNBP3 in diseases

A technology for expressing products and genes, applied in the field of diagnosis and treatment of rectal adenocarcinoma of HNBP3 gene and its expression products, can solve problems such as tumor metastasis

Active Publication Date: 2016-06-08
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, despite great progress in early diagnosis and comprehensive treatment, tumor metastasis and recurrence are still the primary reasons affecting prognosis

Method used

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  • Gene HRNBP3 and new application of expression product of gene HRNBP3 in diseases
  • Gene HRNBP3 and new application of expression product of gene HRNBP3 in diseases
  • Gene HRNBP3 and new application of expression product of gene HRNBP3 in diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 Screening for gene markers associated with rectal adenocarcinoma

[0058] 1. Collection of samples

[0059] 8 cases of rectal adenocarcinoma paracancerous tissues and rectal adenocarcinoma tissue samples were collected, and all the above samples were obtained with the consent of the organizational ethics committee.

[0060] 2. Preparation of RNA samples (operated using QIAGEN tissue RNA extraction kit)

[0061] 1) Tissue extraction

[0062] In a clean area with less RNase interference, use a mortar containing an appropriate amount of liquid nitrogen to weigh the isolated intervertebral disc nucleus pulposus tissue sample, grind it to powder with a pestle, and then transfer the sample to a RNase-free 2ml centrifuge tube middle. Add 300 μl of lysate, place in a homogenizer, grind thoroughly for 5 minutes, centrifuge at 12,000 g at 4°C for 10 minutes, and transfer the supernatant to a new 1.5ml centrifuge tube. Add 600 μl of RNase-free water, mix with a vortex,...

Embodiment 2

[0080] Example 2 QPCR sequencing to verify the differential expression of the HRNBP3 gene

[0081] 1. According to the detection results of high-throughput sequencing, the HNBP3 gene was selected for large-sample QPCR verification. According to the sample collection method in Example 1, 70 cases of rectal adenocarcinoma paracancerous tissues and 70 cases of rectal adenocarcinoma tissues were selected.

[0082] 2. The RNA extraction steps are the same as in Example 1.

[0083] 3. Reverse transcription: use the reverse transcription kit of TAKARA company to operate. Specific steps are as follows:

[0084] (1) Take 1 μg of total RNA for reverse transcription, add 2 μl of Oligo(dT), and mix well. Immediately after 5 min in water bath at 70°C, ice bath for 2 min.

[0085] (2) Construct a 25 μl reaction system, including 5 μl of 5× reverse transcription buffer, 5 μl of dNTP (2.5 mM), 40 U / μl of RNasin, 200 U / μl of M-MLV, and make up to the expected volume with nuclease-free wate...

Embodiment 3

[0106] Inhibition of embodiment 3HRNBP3 gene expression

[0107] 1. Cell culture: human rectal adenocarcinoma cell line HRC-99, cultured in RPMI1640 medium containing 10% calf serum and 1% P / S at 37°C, 5% CO 2 , Cultivated in an incubator with a relative humidity of 90%. Change the medium once every 2-3 days, and use 0.25% EDTA-containing trypsin for routine digestion and passage.

[0108] 2. siRNA design

[0109] siRNA sequences against HNBP3:

[0110] siRNA1-HRNBP3:

[0111] The sense strand is 5'-AGUUACAAACCCAAAACCCUU-3' (SEQ ID NO.7),

[0112] The antisense strand is 5'-GGGUUUUGGGUUUGUAACUUU-3' (SEQ ID NO.8);

[0113] siRNA2-HRNBP3:

[0114] The sense strand is 5'-AGUUUCAAAAGUUACAAACCC-3' (SEQ ID NO.9),

[0115] The antisense strand is 5'-GUUUGUAACUUUUGAAACUAG-3' (SEQ ID NO.10);

[0116] siRNA3-HRNBP3:

[0117] The sense strand is 5'-AUUAUUGACCUCAAUUUUCCG-3' (SEQ ID NO.11),

[0118] The antisense strand is 5'-GAAAAUUGAGGUCAAUAAUGC-3' (SEQ ID NO.12);

[0119] Neg...

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Abstract

The invention discloses a gene HRNBP3 and new application of an expression product of the gene HRNBP3 in diseases, and particularly relates to the gene HRNBP3 and application of the expression product of the gene HRNBP3 in diagnosis and treatment of the rectal adenocarcinoma. The invention further discloses application of the gene HRNBP3 in preparation of a product for diagnosing the rectal adenocarcinoma, and meanwhile discloses application of the gene HRNBP3 in preparation of medicine for treating the rectal adenocarcinoma.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a new application of the HRNBP3 gene and its expression product in diseases, in particular to a new application of the HRNBP3 gene and its expression product in the diagnosis and treatment of rectal adenocarcinoma. Background technique [0002] Abroad, rectal adenocarcinoma is one of the common malignant tumors of the digestive tract, and its incidence rate has been at a relatively high level. In China, with the changes in people's lifestyle and diet, the incidence of rectal adenocarcinoma is increasing year by year. In some southern cities, the age of onset of rectal adenocarcinoma tends to be younger, seriously endangering human health. The pathogenic factors of rectal adenocarcinoma are diverse and complex, and are the result of multi-factor and multi-step development, but the specific pathogenesis has not been clarified so far. At present, it is believed that the pathogenesis of r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K31/7088A61K48/00A61P35/00
CPCA61K31/7088C12Q1/6886C12Q2600/158G01N33/57446G01N33/68
Inventor 杨承刚常鹏孙耀兰
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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