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Method of inhibiting proliferation of ovarian cancer cells

An ovarian cancer cell, phosphorylation level technology, applied in tumors/cancer cells, animal cells, other methods of inserting foreign genetic materials, etc., can solve the problems of difficult tumor proliferation and growth, unsatisfactory treatment effect, and low response rate.

Inactive Publication Date: 2019-12-13
BEIJING CHAOYANG HOSPITAL CAPITAL MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The biggest problem facing chemotherapy is chemotherapy resistance. After drug resistance, it is difficult for chemotherapy drugs to control the proliferation and growth of tumors.
Ovarian cancer drug resistance has become one of the main problems that plague clinical treatment. The response rate of drug-resistant and refractory ovarian cancer to retreatment is very low, only 10-15%. The treatment effect is not ideal. At present, only A palliative treatment plan focused on relieving symptoms

Method used

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  • Method of inhibiting proliferation of ovarian cancer cells
  • Method of inhibiting proliferation of ovarian cancer cells
  • Method of inhibiting proliferation of ovarian cancer cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1, the expression level of ATAD2 protein in ovarian cancer tissue (primary ovarian cancer tissue or gastric cancer metastasis ovarian cancer tissue) is significantly up-regulated

[0054] The ovarian cancer tissue chip (Alenabio Company (Xi'an, China)) contained 60 primary ovarian cancer tissues (i.e., primary lesions), 10 ovarian cancer tissues with gastric cancer metastases (i.e., gastric metastases), and 10 adjacent normal tissues.

[0055] 1. The ovarian cancer tissue chip was detected by IHC standard method to evaluate the expression of ATAD2 protein in ovarian cancer tissue. The specific steps are as follows:

[0056] (1) Dewaxing and hydration

[0057] The paraffin sections were placed in an oven at 60°C and baked for 60 minutes. After the sections were taken out, they were dewaxed three times in xylene I, xylene II, and xylene III, each time for 10 minutes. Then put them into absolute ethanol, 95% ethanol aqueous solution, 90% ethanol aqueous solution,...

Embodiment 2

[0077] Example 2. Using public database data to verify the up-regulated expression of ATAD2 protein in ovarian cancer tissues

[0078] 1. In order to verify the up-regulated expression of ATAD2 protein in ovarian cancer tissues, the inventors of the present invention evaluated the CSIOVDB database (Tan TZ, Yang H, Ye J, et al. CSIOVDB: a microarray gene expression database of epithelial ovarian cancer subtype. Oncotarget .2015.6(41):43843-52.) The expression level of ATAD2mRNA in ovarian tissue (including ovarian cancer tissue and normal ovarian tissue). The CSIOVDB database includes transcriptome data of 3431 ovarian cancer tissues and normal ovarian tissues.

[0079] See the experimental results figure 2 Middle A.

[0080] 2. In order to verify whether the expression of ATAD2mRNA in ovarian cancer tissue is higher than the normal level, the inventors of the present invention use the GEO2R online tool (URL: https: / / www.ncbi.nlm.nih.gov / geo / geo2r / ) , and further analyzed t...

Embodiment 3

[0083] Example 3, ATAD2 gene copy number variation leads to up-regulation of ATAD2 gene expression

[0084] 1. Putative copy number calls determined for the TCGA dataset using GISTIC 2.0 for copy number variation (CNV) analysis (Mermel CH, Schumacher SE, Hill B, Meyerson ML, Beroukhim R, GetzG. GISTIC2.0 facilitates sensitive and confident localization of the targets of focal somatic copy-number alteration in human cancers. Genome Biol. 2011.12(4):R41.). Copy number: -2 = homozygous deletion; -1 = hemizygous deletion; 0 = neutral / no change; 1 = gain; 2 = amplification.

[0085] 2. Using Oncomine analysis (Rhodes DR, Yu J, Shanker K, et al. ONCOMINE: a cancer microarray database and integrated data-mining platform. Neoplasia. 2004.6 (1): 1-6.) for 607 cases of ovarian cancer in the TCGA dataset Tissues, 130 cases of normal ovarian tissue and 431 pairs of normal blood samples were compared.

[0086] See the experimental results image 3 Middle A. The results showed that the ...

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Abstract

The invention discloses a method for inhibiting proliferation of ovarian cancer cell. According to the method, a CRISPR / Cas9 system is utilized to edit an ATAD2 gene in an ovarian cancer cell (a cellstrain SKOV3 or a cell strain A2780) genome, so that the expression quantity and / or activity of ATAD2 protein are / is reduced; wherein the CRISPR / Cas9 system comprises sgRNA1 and sgRNA2, and a target site recognized by the sgRNA1 is a DNA molecule shown in a sequence 1 in a sequence table; wherein the target site recognized by the sgRNA2 is the DNA molecule shown as a sequence 2 in the sequence table. The experiments prove that compared with control ovarian cancer cells, the proliferation ability, the growth ability and the clone formation ability of the ovarian cancer cells with ATAD2 genes knocked out are all remarkably reduced, and the important significance is achieved for ovarian cancer treatment. The method has important application value.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a method for inhibiting the proliferation of ovarian cancer cells. Background technique [0002] Ovarian cancer is a tumor of the female reproductive system, accounting for 2.5% of all malignant tumors in women, but due to low survival rate, its mortality rate accounts for 5% of female cancer deaths. An estimated 22,240 new cases of ovarian cancer and 14,070 deaths from ovarian cancer were estimated to have occurred in the United States in 2018. Despite some advances in surgical techniques and adjuvant therapy, survival rates for ovarian cancer have changed little. Therefore, there is an urgent need to discover more precise oncogenic mechanisms and to develop new effective treatments to improve the survival of ovarian cancer patients. [0003] The current main methods of controlling the growth of ovarian cancer cells include surgical treatment and chemotherapy. Surgery i...

Claims

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Application Information

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IPC IPC(8): C12N15/867C12N15/90C12N5/10C12N9/22A61K48/00A61P35/00
CPCA61K48/005A61P35/00C12N5/0693C12N9/22C12N15/86C12N15/907C12N2510/00C12N2740/15043
Inventor 刘健刘群
Owner BEIJING CHAOYANG HOSPITAL CAPITAL MEDICAL UNIV
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