CN113796349A - Animal model for attacking nerves and inducing pain of breast cancer and preparation method thereof - Google Patents
Animal model for attacking nerves and inducing pain of breast cancer and preparation method thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract
The invention discloses an animal model for attacking nerves and inducing pain of breast cancer and a preparation method thereof, belonging to the technical field of medicines. According to the invention, a 4T1 breast cancer cell strain is selected, the tumor cell is injected under the mouse sciatic nerve epitoma, the change of the sciatic nerve functional index and pain sensation of the injected mouse is observed, an effective model of the breast cancer invasive nerve is initially established, and a certain experimental basis is laid for further deeply discussing the PNI action of the breast cancer and the mechanism causing pain; has important significance for early diagnosis and treatment of breast cancer, improving treatment effect and relieving pain of clinical breast cancer patients, especially for preventing and relieving PMPS.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to an animal model for invading nerves and inducing pain of breast cancer and a preparation method thereof.
Background
World health international agency for research on cancer (IARC) release in 2020: breast cancer has become the first cancer to develop worldwide in place of lung cancer. Breast cancer is a malignant tumor seriously harming the health of women, and relapse and metastasis are main reasons of treatment failure of the breast cancer. Neurotropic Invasion (PNI) is the 5 th mode of metastasis following direct spread, colonization, lymphatic metastasis and hematogenous metastasis, playing an important role in malignant tumor invasion. Research shows that PNI is closely related to recurrence and 5-year survival rate of patients with invasive breast cancer, and is an important prognostic parameter of breast cancer. However, the study on the mechanism of nerve invasion is slow due to the lack of a stable and effective in vivo PNI model.
In addition, early stage breast cancer does not have any symptoms and pain. However, if there is lymph node metastasis, brain metastasis or bone metastasis at the late stage, pain of different degrees often appears according to the metastatic site; chronic neuropathic pain (PMPS) occurs in 13% -53% of breast cancer patients after surgery. These types of pain are a medical, quality of life, and psychosocial stress on many breast cancer patients.
Therefore, the search and preparation of a suitable breast cancer PNI animal model for deeply discussing the mechanism of infiltrating nerves and inducing pain of breast cancer have important medical significance and profound social significance.
Disclosure of Invention
In view of the above-mentioned deficiencies of the prior art, the present invention aims to provide an animal model for invasion of nerves and pain induction of breast cancer and a preparation method thereof.
The invention selects 4T1 breast cancer cells, and the tumor cells are injected under the mouse sciatic nerve epitoma to observe the change of sciatic nerve functional index and pain sensation of the injected mouse, and an effective model of the breast cancer invasive nerve is initially established, so as to lay a certain experimental basis for further deeply discussing the PNI action of the breast cancer and the mechanism causing pain; has important significance for early diagnosis and treatment of breast cancer, improving treatment effect and relieving pain of clinical breast cancer patients, especially for preventing and relieving PMPS.
In one embodiment of the present invention, the 4T1 breast cancer cells are administered in the form of a cell suspension prepared as follows: digesting the cultured 4T1 breast cancer cells with 0.25% trypsin to prepare single cell suspension, washing with normal saline, and suspending with normal saline to obtain 4T1 breast cancer cell suspension.
Further, the cell concentration of the 4T1 breast cancer cell suspension is 1X 107one/mL.
Furthermore, the 4T1 breast cancer cell suspension is injected with the dosage of 5 mu L, and is locally injected under the sciatic nerve epididymis of Balb/c mice.
The preparation method of the breast cancer model for attacking nerves and inducing pain comprises the following specific steps:
mice were shaved, opened along the ischial spaces, and fascia was bluntly isolated using 1.5 doses of isoflurane, aeroma Balb/c. Care was taken to free the sciatic nerve with forceps. The 4T1 tumor cells were evenly blown by a 200. mu.l pipette about 10 times, 5. mu.l of tumor cells were aspirated by a syringe, and the parallel needle was inserted into the sciatic nerve adventitia. The cells are kept for 10s after injection, so that the liquid is fully absorbed, and the suture is performed after the needle is slowly pulled out.
The invasive nerve growth starts 1 week after the injection of the tumor cells, and the tumor body size of each mouse is basically consistent. After 1 week of injection, the sciatic nerve function index and pain threshold caused by tumor compression or infiltration change remarkably, the difference is more remarkable at 2w, and after 3 weeks, the nerve is infiltrated and separated by tumor cells and does not feel pain stimulation any more.
The invention relates to an animal model for the invasion of nerves and the induction of pain of breast cancer, which is prepared by using 4T1 breast cancer cells for the first time.
The animal model for the invasion of nerves and the induction of pain of breast cancer has the advantages of low cost, high speed, good stability, repeatability and worth popularization.
Drawings
FIG. 1 shows the photo-microscopic morphology (X200) of mouse 4T1 breast cancer cells in example 1 cultured to 90% density.
FIG. 2 is the result of the growth of infiltrating nerves 15 days after the injection of tumor cells in example 1.
FIG. 3 Effect of the sub-epidermal injection of 4T1 cells into the sciatic nerve of mice in example 1 on pain from needle puncture of the sole (left) and thermal stimulation (right).
FIG. 4 is the HE staining of sciatic nerve sections of 15d tumor infiltration growth after the mice in example 1 were injected with 4T1 cells under the sciatic nerve membrane. Wherein: the left image is in the form of a macroscopic slice; the right image is the high power under-lens slice morphology.
Detailed Description
The invention is described in further detail below with reference to the figures and the specific examples, which should not be construed as limiting the invention. Modifications or substitutions to methods, procedures, or conditions of the invention may be made without departing from the spirit and scope of the invention. The experimental methods and reagents of the formulations not specified in the examples are in accordance with the conventional conditions in the art.
Example 1
4T1 is a 6-thioguanine mouth-resistant cell strain which is obtained from a 410.4 tumor strain without mutagenesis screening, can grow adherently, and has the growth and transfer characteristics close to those of breast cancer in a human body in a BALB/c mouse. This tumor is an animal model of human stage VI breast cancer. The kinetics of 4T 1-induced tumor metastasis in postoperative and non-postoperative cases are similar and can be used as a model for both postoperative and non-surgical situations.
In the embodiment, a 4T1 breast cancer cell line is selected, the tumor cell is injected under the mouse sciatic nerve epitoma, and the change of the sciatic nerve functional index and pain sensation of the mouse after injection is observed, so that an effective model of the breast cancer invasive nerve is initially established.
The specific process is as follows:
(1) cell culture
Thawing and recovering the cryopreserved 4T1 cell line, placing the cell line in DMEM culture solution containing fetal bovine serum with the volume fraction of 10%, and placing a culture dish inoculated with the cells in CO with the volume fraction of 5%2And culturing in a 37 ℃ incubator with saturated humidity.
The morphology of mouse 4T1 breast cancer cells when cultured to 90% density under a light microscope is shown in fig. 1.
(2) Injection under sciatic nerve outer membrane
Digesting the cultured 4T1 breast cancer cells with 0.25% trypsin to prepare single cell suspension, washing with normal saline, and suspending with normal saline to obtain 4T1 breast cancer cell suspension, wherein the cell concentration in the 4T1 breast cancer cell suspension is 1 × 107one/mL.
Mice were shaved, opened along the ischial spaces, and fascia was bluntly isolated using 1.5 doses of isoflurane, aeroma Balb/c. The sciatic nerve was carefully picked with forceps. The 4T1 tumor cell suspension was evenly blown by a 200. mu.L pipette about 10 times, 5. mu.L of tumor cells were taken and dropped on a PE glove. Tumor cells were aspirated by syringe and passed in parallel into the sciatic nerve. Pumping into cells, and staying for about 10s to prevent liquid from leaking when the injector is pulled out; and (6) sewing.
Testing the sciatic nerve function index (SFI) and pain threshold of the mice at 7d and 14d of injection respectively; and (5) anesthetizing the mice at 15d, exposing sciatic nerves after perfusion, taking out sciatic nerves with long tumors, measuring the sizes of the tumors, and storing the sciatic nerves with the long tumors in a refrigerator at the temperature of-80 ℃ or 4% paraformaldehyde for later use.
FIG. 2 shows the results of infiltrating nerve growth 15 days after tumor cell injection.
(3) Sciatic nerve function assay
Sciatic nerve function index (SFI) test: a wooden groove with the length of 60cm, the width of 10cm and the height of 10cm and two open ends is manufactured, 70g of white paper is cut into the same length and width as the wooden groove, and then the white paper is laid at the bottom of the groove. After coloring the double hind limbs of the rat by soaking the pigment in the double ankle joints, the rat is placed at one end of the groove and walks to the other side of the groove by itself, and 5-6 footprints are left on each hind limb. The footprints with clear footprints were selected to measure 3 indices of normal (N) and injured lateral (E) feet, respectively: A. PL (footprint length); B. TS (toe width); C. IT (medial toe width). Substituting the indexes into Bain formula to calculate sciatic nerve function index. Bain formula: SFI =109.5(ETS-NTS)/NTS-38.3 (EPL-NPL)/NPL +13.3 (EIT-NIT)/NIT-8.8. Sciatic nerve function index SFI =0 is normal, -100 is complete injury.
(4) von frey prick test
Adapting to Day 1-3 mice; day4 was allowed to rest with the mice for a period of time before the measurement and then allowed to rest; the measurements were carried out using 0.16g, 0.4g, 0.6g, 1g, 2g needles, at 5-10s intervals per measurement; when measuring, a 0.16g needle is used for finding the arch of the mouse, after the arch of the mouse is contacted, the needle is slightly bent by force, if no reaction exists, the reaction is carried out, and the mark is multiplied; if the reaction occurs, after the patient is calm, the patient needs to be stimulated by a needle of 0.16g, the reaction lasts for 0.16g, and the measurement is carried out for 4 times, and if the reaction is not painful from 0.16g to 2.0g, the measurement is carried out for five times. If no reaction, changing to 0.4g, and if the reaction occurs, then using 0.16g of needle to stimulate; this was repeated and the measurements were 6 times.
(5) Statistical analysis: all data were statistically analyzed using mean ± standard deviation using SPSS 16.0 software. And (5) adopting a t test. P < 0.05 is statistically significant for the differences.
As shown in fig. 3, the pain threshold of 7d plantar acupuncture after injection of tumor cells into sciatic nerve of mice was significantly reduced, indicating that tumor infiltrating nerve grows and affects pain afferent; the pain threshold of the 7d plantar thermal stimulation of the mouse after the sciatic nerve is injected with tumor cells is obviously reduced, which indicates that the tumor infiltrating nerve grows and influences the transmission of pain sense.
As shown in FIG. 4, the mouse sciatic nerve was injected with 4T1 cells under the outer membrane to invade nerve growth, to thin nerve bundles, and to disorder the arrangement of nerve fibers. There is tumor cell infiltration and growth among nerve fibers.
Claims (6)
1. An animal model of breast cancer affecting nerves and inducing pain, characterized by: the animal model is obtained by locally injecting 4T1 breast cancer cells under the animal sciatic nerve adventitia.
2. The animal model of claim 1, wherein: the animal is a mouse.
3. A method of producing the animal model of claim 1, wherein: the method comprises the following steps:
step 1, anesthetizing a Balb/c mouse, shaving hairs, opening along the ischial gap, carrying out blunt separation on fascia, and dissociating sciatic nerves by using forceps;
and 2, sucking 4T1 breast cancer cell suspension, injecting the breast cancer cell suspension into the sciatic nerve adventitia, and suturing after injection.
4. The production method according to claim 3, characterized in that: the cell concentration of the 4T1 breast cancer cell suspension is 1 x 107one/mL.
5. The method of claim 4, wherein: the injection dosage of the 4T1 breast cancer cell suspension in the step 2 is 5 mu L.
6. The method of claim 4, wherein: the 4T1 breast cancer cell suspension is prepared as follows: digesting the cultured 4T1 breast cancer cells with 0.25% trypsin to prepare single cell suspension, washing with normal saline, and suspending with normal saline to obtain 4T1 breast cancer cell suspension.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102256623A (en) * | 2008-06-05 | 2011-11-23 | 独立行政法人国立癌症研究中心 | Neuroinvasion inhibitor |
| CN103845730A (en) * | 2012-12-05 | 2014-06-11 | 复旦大学 | Application of anti-ST2/IL-1R4 antibody in preparation of pain relieving medicament |
| CN106109495A (en) * | 2016-06-03 | 2016-11-16 | 中山大学附属第五医院 | A kind of live body sciatic method of dynamic tracer tumor cell invasion |
| US9807986B2 (en) * | 2016-03-28 | 2017-11-07 | Rui Hou | Methods for establishing animal model of hepatocellular carcinoma bone metastasis |
| CN108785344A (en) * | 2018-08-22 | 2018-11-13 | 南京中医药大学 | Rhizoma Et Radix Notopterygii is preparing the application in preventing neurogenic pain disease medicament |
| CN110079501A (en) * | 2019-05-14 | 2019-08-02 | 兰州大学 | Mouse breast cancer circulating tumor cell system and its method for building up |
| CN112754717A (en) * | 2021-01-14 | 2021-05-07 | 华中科技大学同济医学院附属协和医院 | Method for building disease model of breast cancer bone metastasis laboratory mouse |
-
2021
- 2021-08-06 CN CN202110900165.4A patent/CN113796349A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102256623A (en) * | 2008-06-05 | 2011-11-23 | 独立行政法人国立癌症研究中心 | Neuroinvasion inhibitor |
| CN103845730A (en) * | 2012-12-05 | 2014-06-11 | 复旦大学 | Application of anti-ST2/IL-1R4 antibody in preparation of pain relieving medicament |
| US9807986B2 (en) * | 2016-03-28 | 2017-11-07 | Rui Hou | Methods for establishing animal model of hepatocellular carcinoma bone metastasis |
| CN106109495A (en) * | 2016-06-03 | 2016-11-16 | 中山大学附属第五医院 | A kind of live body sciatic method of dynamic tracer tumor cell invasion |
| CN108785344A (en) * | 2018-08-22 | 2018-11-13 | 南京中医药大学 | Rhizoma Et Radix Notopterygii is preparing the application in preventing neurogenic pain disease medicament |
| CN110079501A (en) * | 2019-05-14 | 2019-08-02 | 兰州大学 | Mouse breast cancer circulating tumor cell system and its method for building up |
| CN112754717A (en) * | 2021-01-14 | 2021-05-07 | 华中科技大学同济医学院附属协和医院 | Method for building disease model of breast cancer bone metastasis laboratory mouse |
Non-Patent Citations (1)
| Title |
|---|
| 德马亚·巴: "《乳腺癌组学技术 新一代诊断、预后评估和治疗技术》", 31 March 2017, 天津科技翻译出版公司 * |
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