DSTN gene and expression product thereof as diagnosis and treatment target of endometrial cancer

An endometrial cancer, gene technology, applied in the field of cancer diagnosis, prognosis prediction, and treatment, can solve the problems of occult metastasis, high mortality, and poor patient response to radiation therapy.

Inactive Publication Date: 2016-08-24
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In clinical work, although most patients with endometrial cancer first present with postmenopausal vaginal bleeding or irregular vaginal bleeding, and then are diagnosed as early endometrial cancer (stage), there are still more than 100 patients who have been diagnosed at the time of diagnosis. Locally advanced or occult metastases, local or distant metastases have occurred, such patients have poor response to surgery and radiotherapy, and have a high mortality rate

Method used

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  • DSTN gene and expression product thereof as diagnosis and treatment target of endometrial cancer
  • DSTN gene and expression product thereof as diagnosis and treatment target of endometrial cancer
  • DSTN gene and expression product thereof as diagnosis and treatment target of endometrial cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1 Gene chip screening of differentially expressed genes

[0070] 1. Sample collection:

[0071] Tissue samples of endometrial cancer: Collect endometrial cancer patients, all patients were treated with surgery, 10 cases of surgical paraffin specimens. All patients were diagnosed with endometrial cancer after pathological examination. Other entry conditions were: all patients had not received any treatment before admission: no other malignant tumors; no other hormone-related diseases; complete clinical data.

[0072] 10 patients with endometrial cancer, the average age of onset was 58 years old. The main clinical manifestations of the patients are irregular vaginal bleeding, lower abdominal pain, menstrual disorders, vaginal discharge, etc. Some patients have no obvious symptoms but are found on physical examinations. Endometrial cancer specimens were sectioned by HE staining, and histomorphology was confirmed to be endometrial cancer.

[0073] Normal endometrial tiss...

Embodiment 2

[0084] Example 2 Large sample verification and screening of differentially expressed genes

[0085] Considering that there is no gene in the prior art that has been studied on the correlation between this gene and endometrial cancer as a candidate gene, and considering the results of gene sequencing, the DSTN gene (its expression is down-regulated in endometrial cancer tissue) is selected for verification .

[0086] 1. Sample collection

[0087] According to the method of Example 1, 50 cases of endometrial cancer tissue and 60 cases of normal endometrial tissue were collected.

[0088] 2. Validation at the mRNA level

[0089] 2.1 Extract tissue RNA

[0090] The steps are the same as in Example 1.

[0091] 2.2 Reverse transcription

[0092] Using reverse transcription kit, reverse transcription of lμg total RNA with reverse transcription buffer to synthesize cDNA. A 25μl reaction system was used, and 1μg total RNA was used as template RNA for each sample. The following components were add...

Embodiment 3

[0115] Example 3 DSTN gene overexpression

[0116] 1. Plasmid construction

[0117] The amplification primers are designed according to the coding sequence of the DSTN gene, and the design of the primers is well known to those skilled in the art. Amplify the coding sequence of the full-length DSTN gene from the cDNA library of adult fetal brain (clontech company, article number: 638831), insert the above cDNA sequence into the eukaryotic expression vector pcDNA3.1, and ligate the obtained recombinant vector pcDNA3.1 -DSTN is used for follow-up experiments.

[0118] 2. Cultivation and transfection of endometrial cancer cells

[0119] 2.1 Cell culture

[0120] The Ishikawa3-H-12 cell lines were cultured in RPMI 1640 medium (containing 10% fetal bovine serum, 100U / ml penicillin, 100g / ml streptomycin) in 50ml culture flasks adherently, at 37℃, 5% CO 2 Continuous cultivation in an incubator with saturated humidity.

[0121] 2.2 Cell transfection

[0122] 1. 0.5-2*10 the day before transfecti...

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Abstract

The invention discloses a DSTN gene which can be used as a molecular marker of endometrial cancer diagnosis. A gene chip and a QPCR experiment are utilized to find that the DSTN gene and a coding protein thereof have remarkable difference in expression in a normal endometrial tissue and an endometrial cancer tissue, namely that the expression condition of the DSTN gene in the endometrial tissue can be detected to judge whether a subject suffers from endometrial cancer or has the risk of suffering from endometrial cancer or not. According to the correlation of the two, the invention develops a kit for diagnosing the endometrial cancer, and the kit is used for diagnosing the endometrial cancer by detecting the expression of the DSTN gene. The diagnosis kit can be used for early-stage diagnosis of diseases, and has a wide application prospect clinically. Besides, the invention further discloses the DSTN gene which can be used as a molecular target for treating the endometrial cancer, so that a foundation is provided for developing an endometrial cancer treatment drug.

Description

Technical field [0001] The present invention relates to the field of cancer diagnosis, treatment, and prognosis prediction. More specifically, the present invention relates to a method for cancer diagnosis and prognosis prediction by detecting DSTN abnormalities; and a cancer therapeutic agent that activates DSTN genes or proteins. Background technique [0002] Globally, endometrial cancer (EC) is one of the most common malignant tumors of the female reproductive system. Epidemiological data show that endometrial cancer ranks fourth in all system malignancies in women in developed countries (after breast, colorectal cancer and lung cancer) and the first in reproductive system malignancies (accounting for all More than 50% of new cases of gynecological reproductive system tumors). According to statistics, the number of new cases of endometrial cancer in the United States in 2014 will reach 52,630, and 8590 patients are expected to die from the disease. Similarly, the incidence o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61P35/00
CPCC12Q1/6886A61K45/00C12Q2600/118C12Q2600/158G01N33/57442G01N33/68G01N2333/47
Inventor 杨承刚宋宏涛
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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