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Application of linc00540 in the diagnosis and treatment of rectal adenocarcinoma

A rectal adenocarcinoma, application technology, applied in the application field of LINC00540 in the diagnosis and treatment of rectal adenocarcinoma, can solve the problem of no significant improvement in the prognosis of colorectal cancer patients

Active Publication Date: 2020-08-11
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, although diagnosis and treatment techniques and chemotherapy drugs have been continuously updated, the treatment effect of colorectal patients has been greatly improved, but the prognosis of colorectal cancer patients has not been significantly improved because most patients have metastases when they are found (Anaya DA, Becker NS ,Abraham NS.Globalgraying,colorectal cancer and liver metastasis:new implications for surgical management[J].Crit Rev Oncol Hematol 2011,77(2):100-8.)

Method used

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  • Application of linc00540 in the diagnosis and treatment of rectal adenocarcinoma
  • Application of linc00540 in the diagnosis and treatment of rectal adenocarcinoma
  • Application of linc00540 in the diagnosis and treatment of rectal adenocarcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1 Screening for gene markers associated with rectal adenocarcinoma

[0068] 1. Sample collection

[0069] 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.

[0070] 2. Preparation of RNA samples

[0071] Tissue RNA was extracted using QIAGEN tissue RNA extraction kit, and the specific operation was carried out according to the instructions.

[0072] 3. Total RNA quantification and purity analysis

[0073] The above extracted RNA was subjected to agarose gel electrophoresis, the concentration and purity ...

Embodiment 2

[0094] Example 2 QPCR sequencing to verify the differential expression of LINC00540 gene

[0095] 1. Large-scale QPCR verification of the differential expression of the LINC00540 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.

[0096] 2. RNA extraction

[0097] RNA samples were extracted using the QIAGEN Tissue RNA Extraction Kit. For details, please refer to the instruction manual.

[0098] 3. QPCR

[0099] 1) Reverse transcription reaction

[0100] Use FastQμant cDNA First Strand Synthesis Kit (Product No.: KR106) for reverse transcription of mRNA, 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 Enzyme Mix, 2.0μl of FQ-RT Primer Mix, RNase Free ddH 2 O 5.0 μl, after...

Embodiment 3

[0114] Example 3 Silencing of the LINC00540 gene

[0115] 1. Cell culture

[0116] 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%. Change the medium once every 2-3 days, use 0.25% trypsin containing EDTA to routinely digest and passage, and take the cells in the logarithmic growth phase for the experiment.

[0117] 2. Design of siRNA

[0118] siRNA was designed according to the sequence of LINC00540 gene, the sequence of negative control siRNA-NC is shown in SEQ ID NO.5-6; the sequence of siRNA1-4 in the experimental group is shown in SEQ ID NO.7-14.

[0119] 3. Transfection

[0120] Divide the cells into 1×10 4 Inoculate / well into 24-well cell culture plates at 37°C, 5% CO 2 Cells were cultured in an incubator for 24 hours, and transfected in RPMI1640 medium containing no double antibody and containing 10% FBS according to the...

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Abstract

Applications of LINC00540 in rectal adenocarcinoma diagnosis and treatment are disclosed. Whether a patient suffers from rectal adenocarcinoma or not is determined through detecting the expression level of the LINC00540. Rectal adenocarcinoma cell proliferation, migration and invasion can be suppressed through lowering the expression level of the LINC00540, thus providing molecular means for rectal adenocarcinoma treatment.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of LINC00540 in the diagnosis and treatment of rectal adenocarcinoma. Background technique [0002] Rectal adenocarcinoma is a kind of human adenocarcinoma. The cancer body has glandular tube arrangement structure. It can be divided into grades I, II, and III according to the degree of differentiation, and the degree of malignancy varies greatly. Early symptoms of rectal adenocarcinoma include blood in the stool, changes in bowel habits, intestinal stenosis, anal pain, anal incontinence, etc. Anorectal diseases such as hemorrhoids, anal fissure, anal fistula, rectal polyps, and proctitis are the main causes of rectal adenocarcinoma. Rectal adenocarcinoma is divided into saline adenocarcinoma, tubular adenocarcinoma, mucinous adenocarcinoma, undifferentiated carcinoma, adenosquamous carcinoma, and small cell carcinoma. The cancer cells have a typical squamous cell carcinoma...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886A61K45/00A61K31/713A61P35/00
CPCA61K31/713A61K45/00A61P35/00C12Q1/6886C12Q2600/106C12Q2600/136C12Q2600/158C12Q2600/178
Inventor 董东石小峰
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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