A molecular marker associated with the occurrence and development of rectal adenocarcinoma

A rectal adenocarcinoma and material technology, applied in the field of biomedicine, can solve the problems of economic loss of patients' families, incomplete understanding of molecular and genetic mechanisms, and impact on the quality of life of 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, with the clinical application of antibody drugs and anti-angiogenic drugs, the therapeutic effect of rectal cancer has been significantly improved, but the treatment of patients with advanced disease, recurrence after surgery, and resistance to radiotherapy and chemotherapy is still a difficult point in clinical treatment, and the prognosis is still poor. Poor, seriously affecting the quality of life of patients, causing significant economic losses to the patient's family and the National Medical Insurance Fund
Despite the in-depth study of tumors in recent years, the risk factors, pathogenesis, diagnosis, treatment, and prognosis of rectal cancer continue to become research hotspots, but the molecular and genetic mechanisms of rectal cancer are still not completely clear, and the prevention and treatment effect is not ideal. Cancer is still a serious threat to the health and life of the people, and it is a major public health problem worthy of further research.

Method used

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  • A molecular marker associated with the occurrence and development of rectal adenocarcinoma
  • A molecular marker associated with the occurrence and development of rectal adenocarcinoma
  • A molecular marker associated with the occurrence and development of rectal adenocarcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Example 1 Screening for gene markers associated with rectal adenocarcinoma

[0083] 1. Sample collection

[0084] 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.

[0085] 2. Preparation of RNA samples

[0086] Tissue RNA was extracted using QIAGEN tissue RNA extraction kit, and the specific operation was carried out according to the instructions.

[0087] 3. Total RNA quantification and purity analysis

[0088] The above extracted RNA was subjected to agarose gel electrophoresis, the concentration and purity ...

Embodiment 2

[0109] Example 2 QPCR sequencing to verify the differential expression of the LOC105373485 gene

[0110] 1. Large-sample QPCR verification of differential expression of LOC105373485 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.

[0111] 2. RNA extraction

[0112] RNA samples were extracted using the QIAGEN Tissue RNA Extraction Kit. For details, please refer to the instruction manual.

[0113] 3. QPCR

[0114] 1) Reverse transcription reaction

[0115] 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 O5.0 μl, aft...

Embodiment 3

[0135] Example 3 Silencing of the LOC105373485 gene

[0136] 1. Cell culture

[0137] 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.

[0138] 2. Design of siRNA

[0139] The siRNA was designed according to the sequence of the LOC105373485 gene. The sequence of the 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.

[0140] 3. Transfection

[0141] 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 an...

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Abstract

The invention discloses a molecular marker related to genesis and development of rectum adenocarcinoma. A high-throughput sequencing technology and bioinformatics are combined to screen IncRNA representing differential expression in the tissue of rectum adenocarcinoma, which firstly discovers that LOC105373485 represents differential expression in the tissue of rectum adenocarcinoma, furthermore,large sample verification and cell experiments prove that the IncRNA is related to proliferation, migration and invasion of rectum adenocarcinoma.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a molecular marker related to the occurrence and development of rectal adenocarcinoma. The specific molecular marker is LOC105373485. Background technique [0002] Rectal cancer is one of the most common malignancies in the world and an important cause of cancer-related deaths worldwide. The incidence of colorectal cancer shows obvious regional differences in the world, with the highest in North America and lower in Asian and African countries. In recent years, the distribution of the incidence of colorectal cancer in the world has undergone major changes. The incidence in high-incidence regions such as the United States and Europe has declined, and the incidence in traditionally low-incidence countries such as Asia-Pacific has shown a clear upward trend. my country is a country with a low incidence of rectal cancer. With the improvement of my country's economic development level and cha...

Claims

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Application Information

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