CN108103023A - People's carcinoma of the rectum primary cell and its application - Google Patents
People's carcinoma of the rectum primary cell and its application Download PDFInfo
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- CN108103023A CN108103023A CN201810101650.3A CN201810101650A CN108103023A CN 108103023 A CN108103023 A CN 108103023A CN 201810101650 A CN201810101650 A CN 201810101650A CN 108103023 A CN108103023 A CN 108103023A
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- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
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Abstract
The invention discloses people's carcinoma of the rectum primary cell and its application, people's carcinoma of the rectum primary cell behaviour carcinoma of the rectum primary cell HRC 6 is deposited in China typical culture collection center, and deposit number is CCTCC NO：C2017145, people's carcinoma of the rectum primary cell HRC 6 can complete all kinds of molecular biology experiments, and since primary cell experimental result is more reliable compared with cell line experiments result, and closer clinical individual experimental result, therefore the foundation of 6 cells of HRC is of great significance to the morbidity and treatment for studying the Mongolian race (yellow) carcinoma of the rectum.
Description
Technical field
The present invention relates to colorectal cancer tumour cell fields, more particularly to people's carcinoma of the rectum primary cell and its application.
Background technology
The carcinoma of the rectum is common alimentary system malignant tumour, and at present, global carcinoma of the rectum origin can be used for molecular biology
The cell line of experiment only has mono- plant of SW1463, and existing SW1463 cell lines are female Caucasian's kind (white people) origin
Cell line, and cell line molecular biology experiment result and clinical individual result is widely different, it is considered that primary cell is real
It is more reliable compared with cell line experiments result to test result, and closer to clinical individual experimental result, in addition, the culture side of SW1463 cells
Method and most cell line (5%CO2) different, condition of culture is (Air, 100%)；Culture medium is Leibovitz's L-
15Medium, relatively also costly.
The Mongolian race is distributed mainly on China, Korea, Japan, Siberia, South East Asia Mainland, America and Arctic, is
The ethnic group that distribution is most wide in four big ethnic groups, number is most, therefore, the Mongolian race (yellow) carcinoma of the rectum primary cell is built
It is vertical to be of great significance to the morbidity and treatment of studying people's carcinoma of the rectum.
The information for being disclosed in the background section is merely intended to increase the understanding of the general background to the present invention, without answering
When being considered as recognizing or imply that the information structure has been the prior art well known to persons skilled in the art in any form.
The content of the invention
It is an object of the invention to provide people's carcinoma of the rectum primary cell and its application, so as to overcome existing rectum cancer cell
Strain molecule biological experiment result and clinical individual result it is widely different the defects of.
To achieve the above object, the present invention provides a kind of people's carcinoma of the rectum primary cell, people's carcinoma of the rectum primary cells
For people carcinoma of the rectum primary cell HRC-6, China typical culture collection center is deposited in, deposit number is CCTCC NO：
C2017145。
Preferably, in above-mentioned technical proposal, the yield of the carcinomebryonic antigen (CEA) of people's carcinoma of the rectum primary cell HRC-6
It is 0-0.7ng/106Cell/10 day.
Preferably, in above-mentioned technical proposal, the yield of the carcinomebryonic antigen of people's carcinoma of the rectum primary cell HRC-6 is 0.4-
0.6ng/106Cell/10 day.
Preferably, in above-mentioned technical proposal, the tumor associated antigen CA199's of people's carcinoma of the rectum primary cell HRC-6
Expression quantity is 3-19U/106Cell/10 day.
Preferably, in above-mentioned technical proposal, the tumor associated antigen CA199's of people's carcinoma of the rectum primary cell HRC-6
Expression quantity is 13-17U/106Cell/10 day.
Preferably, in above-mentioned technical proposal, when the doubling time of people's carcinoma of the rectum primary cell HRC-6 is 28-32 small.
Preferably, in above-mentioned technical proposal, when the doubling time of people's carcinoma of the rectum primary cell HRC-6 is 30 small.
The present invention also provides application of the above-mentioned people's carcinoma of the rectum primary cell in radiation sensitivity detection.
The present invention also provides application of the above-mentioned people's carcinoma of the rectum primary cell in antitumor drug is prepared.
Compared with prior art, the present invention has the advantages that：
People's carcinoma of the rectum primary cell HRC-6 involved by the application is that the carcinoma of the rectum of the Mongolian race (yellow) origin is primary
Cell, and people's carcinoma of the rectum primary cell HRC-6 can be in 5%CO2Condition of culture under grow, and using culture medium be common base
Basal culture medium RPMI-1640 can also complete all kinds of molecular biology experiments, and since primary cell experimental result is compared with cell
Strain experimental result is more reliable, and closer to clinical individual experimental result, because of building for this person's carcinoma of the rectum primary cell HRC-6 cells
It is vertical, it is of great significance to the morbidity and treatment for studying the Mongolian race (yellow) carcinoma of the rectum.
Preservation information
(i.e. preservation proves " the people's rectum cancer cell embodied to people's carcinoma of the rectum primary cell HRC-6 according to the present invention
HRC-6 ") China typical culture collection center, preservation address have been preserved in August in 2017 within 17th：Chinese Wuhan, Wuhan
University, postcode：430072, deposit number is CCTCC NO：C2017145.
Description of the drawings
Fig. 1 is to amplify 400 under inverted microscope under people's carcinoma of the rectum primary cell HRC-6 cultivation conditions according to the present invention
Photo again；
Fig. 2 is to amplify 400 under inverted microscope after the HE of people's carcinoma of the rectum primary cell HRC-6 according to the present invention is dyed
Photo again；
Fig. 3 is the STR qualification results of people's carcinoma of the rectum primary cell HRC-6 according to the present invention.
Specific embodiment
Below in conjunction with the accompanying drawings, the specific embodiment of the present invention is described in detail, it is to be understood that the guarantor of the present invention
Shield scope is not restricted by specific implementation.
Unless otherwise explicitly stated, otherwise in entire disclosure and claims, term " comprising " or its change
It changes such as "comprising" or " including " etc. and will be understood to comprise stated element or component, and do not exclude other members
Part or other components.
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples, it is unless otherwise specified, commercially commercially available.
1 cell culture of embodiment
(1) tissue pre-treatment：Choose 0.5-1cm3Rectum cancer tissue (rectum cancer tissue source：Sign informed consent form
The First Hospital of Hebei Medical University goes to a doctor patient), sterile saline rinses more than 30s, the excrement on cleaning rectum cancer tissue surface
It becomes point, is immediately placed in that (500U/mL penicillin, 500 μ g/mL streptomysins, 1.25 μ g/mL both sexes are mould containing high concentration antibiotic
Plain B, 80 μ g/mL gentamicins) serum-free RPMI-1640 culture mediums in, after impregnating 5-10 minute, replace fresh containing high concentration
It is impregnated again after the serum-free RPMI-1640 culture mediums of antibiotic 5-10 minutes, in soaking process, overturns and rock frequently, rock, 6-
10 beats/min, preferably to clean；
(2) stripping and slicing：Cancerous tissue is cut into 1-3mm with scalpel3Tissue block；
(3) cultivate：The tissue block cut is put into the culture medium containing antibiotic and carries out initial-stage culture and normal culture, until
Cell attachment：
Initial-stage culture：Contain antibiotic (100-200U/mL penicillin, 100-200 μ g/mL streptomysins, 0.25- with 2ml
0.5 μ g/mL amphotericin Bs) and volume fraction for 20% hyclone RPMI-1640 culture mediums (complete medium) cultivate
When organizing 24-72 small；
Normal culture：After initial-stage culture, with 5ml RPMI-1640 complete medium cultured tissues, cell pastes completely
After wall growth, continue subculture to 2-5 for when, tissue block is removed, only surplus attached cell.
The RPMI-1640 culture mediums of the present invention are the common culture medium in this field.
2 morphological observation of embodiment
By being observed under inverted microscope it can be found that the cell for people's carcinoma of the rectum primary cell that embodiment 1 is turned out
Form be class Epithelial, with large intestine epithelium cell FHC ( CRL-1831TM,https://www.atcc.org/
Products/All/CRL-1831.aspx#characteristics) form is similar, names as people's carcinoma of the rectum primary cell
HRC-6, and China typical culture collection center is preserved within 17th in August in 2017, deposit number is CCTCC NO：
C2017145, concrete shape refer to attached drawing 1；
It is found after people's carcinoma of the rectum primary cell HRC-6 is carried out HE dyeing, the morphological feature with malignant cell,
Cell arrangement is disorderly, loses normal arrangement architecture, cell shape is not of uniform size, refers to attached drawing 2；
People carcinoma of the rectum primary cell HRC-6 is subjected to STR identifications by China typical culture collection center, is as a result confirmed
It is single human origin, and the cell of no cross contamination, concrete outcome refer to attached drawing 3.
3 Biology identification of embodiment
The people carcinoma of the rectum primary cell HRC-6 that above-described embodiment is turned out is passed through into carcinomebryonic antigen detection kit-enzyme-linked
Immunization (Shanghai Jing Dou Bioisystech Co., Ltd) is detected three times, and the yield of the carcinomebryonic antigen detected is respectively
0.4ng/106Cell/10 day, 0.5ng/106Cell/10 day and 0.6ng/106Cell/10 day.
The people carcinoma of the rectum primary cell HRC-6 that above-described embodiment is turned out passes through CA199 detection kits-enzyme linked immunological
Method (Sangon Biotech (Shanghai) Co., Ltd.) is detected three times, the table of the tumor associated antigen CA199 detected
It is respectively 13U/10 up to amount6Cell/10 day, 15U/106Cell/10 day and 17U/106Cell/10 day.
When the doubling time for people's carcinoma of the rectum primary cell HRC-6 that above-described embodiment is turned out is 30 small.
The description of the foregoing specific exemplary embodiment to the present invention is in order to illustrate and illustration purpose.These descriptions
It is not wishing to limit the invention to disclosed precise forms, and it will be apparent that according to the above instruction, can much be changed
And variation.The purpose of selecting and describing the exemplary embodiment is that explain that the certain principles of the present invention and its reality should
With so that those skilled in the art can realize and utilize the present invention a variety of exemplary implementation schemes and
Various chooses and changes.The scope of the present invention is intended to be limited by claims and its equivalents.
Claims (9)
- A kind of 1. people's carcinoma of the rectum primary cell, which is characterized in that people's carcinoma of the rectum primary cell behaviour carcinoma of the rectum primary cell HRC-6, is deposited in China typical culture collection center, and deposit number is CCTCC NO：C2017145.
- 2. people's carcinoma of the rectum primary cell according to claim 1, which is characterized in that people's carcinoma of the rectum primary cell HRC- The yield of 6 carcinomebryonic antigen is 0-0.7ng/106Cell/10 day.
- 3. people's carcinoma of the rectum primary cell according to claim 2, which is characterized in that people's carcinoma of the rectum primary cell HRC- The yield of 6 carcinomebryonic antigen is 0.4-0.6ng/106Cell/10 day.
- 4. people's carcinoma of the rectum primary cell according to claim 1, which is characterized in that people's carcinoma of the rectum primary cell HRC- The expression quantity of 6 tumor associated antigen CA199 is 3-19U/106Cell/10 day.
- 5. people's carcinoma of the rectum primary cell according to claim 4, which is characterized in that people's carcinoma of the rectum primary cell HRC- The expression quantity of 6 tumor associated antigen CA199 is 13-17U/106Cell/10 day.
- 6. people's carcinoma of the rectum primary cell according to claim 1, which is characterized in that people's carcinoma of the rectum primary cell HRC- When 6 doubling time is 28-32 small.
- 7. people's carcinoma of the rectum primary cell according to claim 6, which is characterized in that people's carcinoma of the rectum primary cell HRC- When 6 doubling time is 30 small.
- 8. application of the people's carcinoma of the rectum primary cell described in claim 1 in radiation sensitivity detection.
- 9. application of the people's carcinoma of the rectum primary cell described in claim 1 in antitumor drug is prepared.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810101650.3A CN108103023A (en) | 2018-02-01 | 2018-02-01 | People's carcinoma of the rectum primary cell and its application |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810101650.3A CN108103023A (en) | 2018-02-01 | 2018-02-01 | People's carcinoma of the rectum primary cell and its application |
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| CN201810101650.3A Pending CN108103023A (en) | 2018-02-01 | 2018-02-01 | People's carcinoma of the rectum primary cell and its application |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113278589A (en) * | 2021-06-03 | 2021-08-20 | 北京和合医学诊断技术股份有限公司 | Primary human intestinal cancer cell and culture method and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105154405A (en) * | 2015-10-09 | 2015-12-16 | 河北医科大学第一医院 | Low-injury primary culture method for colorectal cancer cells |
-
2018
- 2018-02-01 CN CN201810101650.3A patent/CN108103023A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105154405A (en) * | 2015-10-09 | 2015-12-16 | 河北医科大学第一医院 | Low-injury primary culture method for colorectal cancer cells |
Non-Patent Citations (2)
| Title |
|---|
| 上海谷研实业有限公司: ""人直肠癌细胞：HRC-6"", 《HTTP://WWW.APP17.COM/C130379/PRODUCTS/D7358798.HTML》 * |
| 郑雪莲等: ""人直肠腺癌细胞系HRC-99的建立及其生物学特性"", 《世界华人消化杂志》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113278589A (en) * | 2021-06-03 | 2021-08-20 | 北京和合医学诊断技术股份有限公司 | Primary human intestinal cancer cell and culture method and application thereof |
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