Application of wfdc21p in the diagnosis and treatment of liver cancer

A technology for WFDC21P and liver cancer, applied in the field of biomedicine, can solve problems such as unclear biological functions

Active Publication Date: 2020-03-13
TAIZHOU MUNICIPAL HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been reported in the prior art that microRNA plays an important role in the development and drug resistance of liver cancer, but the biological function of lncRNA, which is also a non-coding product, and its role in the development of liver cancer are not clear, and further research is needed
[0004] At present, abnormally expressed lncRNAs found in tumor tissues can involve various systems of the body and are widely distributed. However, due to the large number of lncRNAs, the research on lncRNAs in the field of cancer is still in its infancy. Therefore, to explore the molecular basis of lncRNA liver cancer of great significance

Method used

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  • Application of wfdc21p in the diagnosis and treatment of liver cancer
  • Application of wfdc21p in the diagnosis and treatment of liver cancer
  • Application of wfdc21p in the diagnosis and treatment of liver cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1 Screening Gene Markers Related to Liver Cancer

[0070] 1. Sample collection

[0071] The cancer tissues and paracancerous tissues of 10 liver cancer patients were collected, and the patients gave informed consent, and all the above specimens were obtained with the consent of the organizational ethics committee.

[0072] 2. Preparation of RNA samples

[0073] Tissue RNA was extracted using QIAGEN tissue RNA extraction kit, and the operation was performed according to the specific steps in the manual.

[0074]3. Reverse transcription and labeling

[0075] The mRNA was reverse-transcribed into cDNA using the Low RNA Input Linear Amplification Kit, and the experimental group and the control group were labeled with Cy3, respectively.

[0076] 4. Hybridization

[0077] The gene chip uses Cannes Bio-Human lncRNA Array, and hybridization is performed according to the steps of the chip instruction manual.

[0078] 5. Data processing

[0079] After hybridization,...

Embodiment 2

[0082] Example 2 QPCR sequencing to verify the differential expression of WFDC21P gene

[0083] 1. Large-sample QPCR verification of WFDC21P gene differential expression. According to the sample collection method in Example 1, 60 samples of liver cancer tissues and 60 samples of paracancerous tissues were collected.

[0084] 2. The RNA extraction steps are the same as in Example 1.

[0085] 3. Reverse transcription:

[0086] Use 25μl reaction system, take 1μg total RNA for each sample as template RNA, and add the following components to PCR tubes: DEPC water, 5× reverse transcription buffer, 10mM dNTP, 0.1mM DTT, 30μM Oligo dT, 200U / μl M-MLV, template RNA. Incubate at 42°C for 1h, then centrifuge briefly at 72°C for 10min.

[0087] (3) QPCR amplification test

[0088] Primer design:

[0089] The primer sequence of WFDC21P gene is:

[0090] Forward primer: 5'-TCTATTGACTGAAGTTGAC-3' (SEQ ID NO.2)

[0091] Reverse primer: 5'-ATTATGTCTCTGGGTCTT-3' (SEQ ID NO.3)

[0092] T...

Embodiment 3

[0103] Example 3 Differential expression of WFDC21P gene in liver cancer cell lines

[0104] 1. Cell culture

[0105] Human liver cancer cell lines HepG2, Huh7 and normal liver cell line HL-7702 were incubated in DMEM containing 10% fetal bovine serum and 1% P / S at 37°C and 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.

[0106] 2. Extraction of RNA

[0107] 1) Digest the adherent cells with trypsin, and the cells obtained by pipetting are centrifuged, resuspended, washed, and then resuspended in DMEM medium containing 10% FBS;

[0108] 2) Transfer the resuspended cells to a 6-well plate, add medium to 2m1 / well, and shake the 6-well plate to resuspend the cells evenly;

[0109] 3) The cells were adhered to the wall for 48 hours, and the culture medium was removed;

[0110] 4) Lyse the cells with 1ml Trizol reagent, blow and beat the wall o...

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Abstract

The invention discloses an application of WFDC21P in diagnosis and treatment of liver cancer. It is discovered for the first time that a WFDC21P gene achieves up-regulated expression in patient with the liver cancer, and by changing a transcription level of the WFDC21P, proliferative and colony forming units of liver cancer cells can be changed, indicating that the WFDC21P can be used for developing products for early diagnosis of the liver cancer and for developing medicines for treating the liver cancer. Meanwhile, the invention can offer a theoretical basis for researches of a liver cancer molecular mechanism.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of WFDC21P in the diagnosis and treatment of liver cancer. Background technique [0002] Liver cancer is one of the most common malignant tumors and one of the leading causes of cancer-related death worldwide. Eighty percent of new liver cancer cases occur in developing countries, and hepatocellular carcinoma (HCC) is one of the leading causes of cancer death in China. Although local treatments such as surgery and transcatheter arterial chemoembolization can treat HCC, patients with HCC still have a high rate of recurrence and metastasis. The molecular mechanism of HCC recurrence or metastasis is still not very clear. Epithelial-mesenchymal transition (EMT) and mesenchymal-epithelial transition (MET) play an important role in the process of cancer metastasis. In particular, it has been found that EMT may be an important cause of tumor metastatic dissemination and acquis...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886A61K45/00A61K48/00A61K31/713A61P35/00
CPCA61K31/713A61K45/00C12Q1/6886C12Q2600/136C12Q2600/158
Inventor 王冬国陈佳玉杨林军
Owner TAIZHOU MUNICIPAL HOSPITAL
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