Application of lncRNA in the diagnosis and treatment of rectal adenocarcinoma

A rectal adenocarcinoma, expression level technology, applied in the field of biomedicine, can solve the problem of undiscovered lncRNA

Active Publication Date: 2020-08-04
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although lncRNA plays an important role in the occurrence and development of rectal adenocarcinoma, and some lncRNAs can be used as indicators for the diagnosis and prognosis of rectal adenocarcinoma, there are still a large number of lncRNAs that have not been discovered, and further research on the expression of lncRNA in rectal cancer and its regulation of tumor developing mechanism

Method used

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  • Application of lncRNA in the diagnosis and treatment of rectal adenocarcinoma
  • Application of lncRNA in the diagnosis and treatment of rectal adenocarcinoma
  • Application of lncRNA in the diagnosis and treatment of rectal adenocarcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090] Example 1 Screening for gene markers associated with rectal adenocarcinoma

[0091] 1. Sample collection

[0092] Samples of normal epithelial tissue adjacent to rectal adenocarcinoma and tissue samples of rectal adenocarcinoma were collected from 3 cases. All cases had not received chemotherapy and radiotherapy before surgery, and there were no other neoplastic diseases, autoimmune diseases and severe chronic diseases. Normal epithelial tissue adjacent to cancer Tissues were taken from 5cm from the upper edge of the tumor, and all patients gave informed consent, which was approved by the organizational ethics committee.

[0093] 2. Preparation of RNA samples

[0094] Tissue RNA was extracted using QIAGEN tissue RNA extraction kit, and the specific operation was carried out according to the instructions.

[0095] 3. Total RNA quantification and purity analysis

[0096] The above extracted RNA was subjected to agarose gel electrophoresis, the concentration and purity ...

Embodiment 2

[0117] Example 2 QPCR sequencing to verify the differential expression of the LOC105374343 gene

[0118]1. Large-sample QPCR verification of differential expression of LOC105374343 gene. According to the sample collection method in Example 1, 45 cases of normal tissues adjacent to rectal adenocarcinoma and 45 cases of rectal adenocarcinoma tissues were selected.

[0119] 2. RNA extraction

[0120] RNA samples were extracted using the QIAGEN Tissue RNA Extraction Kit. For details, please refer to the instruction manual.

[0121] 3. QPCR

[0122] 1) Reverse transcription reaction

[0123] Use FastQμant cDNA First Strand Synthesis Kit (Product No.: KR106) for lncRNA reverse transcription, first remove the genomic DNA reaction, add 5×gDNA Bμffer 2.0μl, total RNA 1μg, add RNase Free ddH 2 O Make the total volume to 10μl, heat in a water bath at 42°C for 3min, then add 2.0μl of 10×Fast RT Bμffer, 1.0μl of RT EnzymeMix, 2.0μl of FQ-RT Primer Mix, RNase Free ddH 2 O 5.0 μl, after...

Embodiment 3

[0143] Example 3 Silencing of the LOC105374343 gene

[0144] 1. Cell culture

[0145] Human rectal adenocarcinoma cell line HRC-99 was incubated 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%. The medium was changed once every 2-3 days, and 0.25% trypsin containing EDTA was used for routine digestion and passage, and the cells in the logarithmic growth phase were taken for experiments.

[0146] 2. Design of siRNA

[0147] Design siRNA for the sequence of LOC105374343 gene, the designed siRNA sequence is as follows:

[0148] Sequence of negative control siRNA-NC:

[0149] Sense strand: 5'-UUCUCCGAACGUGUCACGU-3' (SEQ ID NO.5),

[0150] Antisense strand: 5'-ACGUGACACGUUCGGAGAA-3' (SEQ ID NO.6);

[0151] siRNA1:

[0152] Sense strand: 5'-UUUUGUUUGCUUCAAGAUGUU-3' (SEQ ID NO.7),

[0153] Antisense strand: 5'-CAUCUUGAAGCAAACAAAAUG-3'(SEQ ID NO.8);

[0154] siRNA2:

[0155] Sense strand: 5...

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Abstract

The invention discloses an application of lncRNA in rectum adenocarcinoma diagnosis and treatment. lncRNA is LOC105374343. The invention first finds that the expression of LOC105374343 in rectum adenocarcinoma is obviously up-regulated, and a large sample test further proves that LOC105374343 can be used as a potential molecular target in rectum adenocarcinoma diagnosis and treatment. The invention also discloses the correlation between LOC105374343 and multiplication, migration and invasion of rectum adenocarcinoma cells and indicates that LOC105374343 can be used as the potential molecular target in clinic treatment of rectum adenocarcinoma.

Description

technical field [0001] The invention belongs to the field of biomedicine, relates to the application of lncRNA in the diagnosis and treatment of rectal adenocarcinoma, and specifically relates to the application of LOC105374343 in the diagnosis and treatment of rectal adenocarcinoma. Background technique [0002] Rectal cancer seriously affects human health and quality of life. Worldwide, colorectal cancer accounts for the third cause of cancer death (Tome LA, Bray F, Siegel RL, et al.Global cancer statistics, 2012[J].CACancer J Clin,2015,65(2):87- 108.). Metastasis and recurrence are the main causes of death in colorectal cancer patients (XiaoYC, Yang ZB, Cheng XS, et al. CXCLB, overexpressed in colorectal cancer, enhances the resistance of colorectal cancer cells to anoikis[J]. Cancer Lett 2015, 361(I ):22-32.). The data provided by the American Joint Committee on Cancer (AJCC) shows that the 5-year survival rate of patients with stage I rectal cancer is 93.2%, while th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886A61K31/7105A61K48/00A61P35/00
CPCA61K31/7105A61P35/00C12Q1/6886C12Q2600/136C12Q2600/178
Inventor 董东石小峰
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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