Molecular marker for diagnosing and treating endometrial cancer

An endometrial cancer, substance technology, applied in the field of cancer diagnosis, prognosis prediction and treatment, can solve the problem of unclear pathogenesis and threat to women's health, etc.

Active Publication Date: 2016-10-12
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the rapid growth of EC cases in my country is consistent with that in foreign countries, which seriously threatens women's health, but its pathogenesis is still unclear

Method used

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  • Molecular marker for diagnosing and treating endometrial cancer
  • Molecular marker for diagnosing and treating endometrial cancer
  • Molecular marker for diagnosing and treating endometrial cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Example 1 Gene Chip Screening for Differentially Expressed Genes

[0074] 1. Sample collection:

[0075] Endometrial cancer tissue samples: Patients with endometrial cancer were collected, all of whom underwent surgical treatment, and 10 surgical paraffin specimens were collected. All patients were diagnosed with endometrial cancer by pathological examination. Other enrollment conditions are: all patients have not received any treatment before admission; no other malignant tumors; no other hormone-related diseases; complete clinical data.

[0076] 10 cases of endometrial cancer patients, the average age of onset was 58 years old. The main clinical manifestations of patients are irregular vaginal bleeding, lower abdominal pain, menstrual disorders, vaginal discharge, etc. There are also some patients who have no obvious symptoms and are found in physical examination. The endometrial cancer specimens were sectioned by HE staining, and the histomorphological diagnosis w...

Embodiment 2

[0088] Example 2 Large sample validation of differentially expressed genes screened out

[0089] Considering the gene that has not been studied in the prior art on the correlation between this gene and endometrial cancer as a candidate gene, and considering the results of gene sequencing, select the SKA3 gene (its expression is up-regulated in endometrial cancer tissue) for verification .

[0090] 1. Sample collection

[0091] According to the method of Example 1, 50 cases of endometrial cancer tissues and 60 cases of normal endometrial tissues were collected.

[0092] 2. Validation at the mRNA level

[0093] 2.1 Extract tissue RNA

[0094] Step is with embodiment 1.

[0095] 2.2 Reverse transcription

[0096] Using a reverse transcription kit, 1 μg of total RNA was reverse-transcribed with reverse transcription buffer to synthesize cDNA. Use 25μl reaction system, take 1μg total RNA for each sample as template RNA, and add the following components in PCR tubes: DEPC wate...

Embodiment 3

[0119] Example 3 Inhibition of SKA3 gene expression

[0120] 1. siRNA design and synthesis

[0121] siRNA sequences against SKA3:

[0122] The sense strand is 5'-UCCAUUAGUACUUUUGUUGCC-3' (SEQ ID NO.5)

[0123] The antisense strand is 5'-CAACAAAAGUACUAAUGGAAA-3' (SEQ ID NO.6);

[0124] The above siRNA sequences and negative control siRNA sequences (siRNA-NC) were provided by Shanghai Gemma Pharmaceutical Technology Co., Ltd.

[0125] 2. Culture and transfection of endometrial cancer cells

[0126] 2.1 Cell culture

[0127] The Ishikawa3-H-12 cell line was cultured in RPMI 1640 medium (containing 10% fetal bovine serum, 100 U / ml penicillin, 100 g / ml streptomycin) in 50 ml culture flasks, at 37 °C, 5% CO 2 Continuous cultivation in a humidified incubator.

[0128] 2.2 Cell transfection

[0129] (1) 24 hours before transfection, inoculate 0.5-2*10 5 cells, the cell confluency was 30-50% at the time of transfection. When plating, the cells should be digested and mixed comp...

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Abstract

The invention discloses that the SKA3 gene can be used as a molecular marker for the diagnosis of endometrial cancer. Experiments prove that the expression quantity of the SKA3 gene in the endometrial cancer tissue is high in comparison with normal endometrial tissue. The invention also discloses that the SKA3 gene also can be used for preparing a medicine for treating endometrial cancer. The research result of the invention provides a new method for clinical diagnosis of endometrial cancer as well as a new target for the medicine for treating endometrial cancer.

Description

technical field [0001] The present invention relates to the fields of cancer diagnosis, treatment, and prognosis prediction. More specifically, the present invention relates to a cancer diagnosis and prognosis prediction method by means of detecting SKA3 abnormality; and a cancer therapeutic agent for inhibiting SKA3 gene or protein. Background technique [0002] Endometrial carcinoma (Endometrial carcinoma EC) is an epithelial malignant tumor that originates in the endometrium, and is one of the three major malignant tumors of female genitalia. In recent years, the incidence of EC has been increasing year by year and tends to be younger. It accounts for about 7% of the total number of female cancers and 20%-30% of malignant tumors of the female reproductive tract. It is close to or even exceeds the incidence of cervical cancer. According to the American Cancer Society (American Cancer Society, ACS) reported that in the United States, EC has become the highest incidence of m...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61P35/00
Inventor 杨承刚宋宏涛
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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