WO2019113899A1

WO2019113899A1 - Anti-tumor related application of gpr1 target and antagonist thereof

Info

Publication number: WO2019113899A1
Application number: PCT/CN2017/116239
Authority: WO
Inventors: 张键; 杨雅莉; 黄斌斌; 代小勇; 陈杰; 肖天霞
Original assignee: 深圳先进技术研究院
Priority date: 2017-12-14
Filing date: 2017-12-14
Publication date: 2019-06-20

Abstract

The present invention provides for an anti-tumor related application of a GPR1 target and an antagonist thereof. The present invention discloses a use of an antagonist of a GPR1 gene or a GPR1 protein for the preparation of a medicament for preventing or treating tumors or their proliferation and migration. Also disclosed is a use of GPR1 gene or GPR1 protein as a target for the prevention and/or treatment of tumors, or as a tumor diagnostic target, or as a drug target for treating or preventing tumors.

Description

GPR1 target and its antagonists and anti-tumor related applications

Technical field

The invention belongs to the field of biomedicine, and particularly relates to the application of GPR1 and its antagonists in anti-tumor.

Background technique

Chemerin acts as a chemokine, chemotaxis of dendritic cells and macrophages, and acts as a bridge between immune and adaptive immunity. On the other hand, as a new adipokines, secreted by adipose tissue, regulates fat cells. Differentiation and lipolysis can promote biological effects such as insulin signaling pathway in adipocytes. More and more studies have shown that the signal abnormality of Chemerin and its receptors is closely related to the occurrence and development of cancer. Chemerin expression level is very low in most tumors, and some are not even detected, but in adjacent tissues. Chemerin is expressed at a higher level in normal tissues at the distal end.

As one of the three receptors of Chemerin, GPR1 belongs to the superfamily of G protein-coupled receptors (GPCRs) and has seven transmembrane helix (7TM) structures, which are expressed in both fat and gonadal tissues. GPCRs play an important role in extracellular signaling to intracellular transduction, regulating cell movement, growth, and gene transcription—a critical factor in these three cancer biology. In the past decade or so, the mediated signaling pathway has been shown to be a key regulator of protooncogene signaling and is a good drug target. Currently, 60% of the drugs sold on the market are designed for this receptor. . But at present, there is still no drug that directly targets GPR1 to treat clinical cancer. Therefore, targeting GPR1, screening new anti-cancer peptide drugs and humanized antibodies has important social significance and broad economic market.

Choriocarcinoma

Choriocarcinoma is a highly malignant tumor that occurs in the epithelial cells of the outer layer of the placenta (ie, trophoblast cells). Secondary to hydatidiform mole, abortion or full-term delivery. The incidence rate is 0.0001% to 0.36%, and a small number can occur after ectopic pregnancy. Choriocarcinoma damages the normal function of the ovaries and seriously jeopardizes the reproductive health of women of childbearing age. There are many ways to infect choriocarcinoma, which can be caused by the symptoms of adjacent organs, such as the appendix. Can also pass lymphatics, blood line spread, etc., sexual contact, not pay attention to hygiene can also cause choriocarcinoma, if not treated in time, can evolve into chronic choriocarcinoma, obstruction of the fallopian tube, leading to ectopic pregnancy or infertility.

The treatment of choriocarcinoma is mainly chemotherapy, supplemented by surgery. Most of the chemical drugs are treated. In the early cases, a single drug can be used, and 5-Fu is the first choice. If the condition is urgent or has reached the advanced stage, two or two kinds are needed. The above drugs are used together. Commonly used is 5-fluorouracil (5-Fu) plus dactinomycin (ksm). 5-Fu and ksm have the best curative effect, and have few side effects and are effective for the transfer of lung, digestive tract, urinary tract and reproductive tract. It can be used for intravenous administration, arterial perfusion, intraluminal or intratumoral injection, or oral administration.

In the case of large lesions, it is estimated that chemotherapy can not be completely treated or slow HCG decline during treatment, uterine perforation intrahepatic metastases, etc., in order to save the lives of patients, surgery is still an important method for the treatment of choriocarcinoma. General hysterectomy and bilateral accessory omentum and parametrial venous plexus and ovarian venous plexus resection.

Breast Cancer

Breast cancer is the world's second-highest mortality fatal cancer after lung cancer. The age-standardized incidence rate of breast cancer in Chinese women is 21.6/100,000, ranking first in female cancer; the mortality rate is 5.7/100,000. , ranked sixth in female cancer deaths. 4% to 6% of breast cancers are metastatic breast cancer at the time of diagnosis, and 30% to 40% of early patients receiving adjuvant therapy can develop metastatic breast cancer, and the 5-year survival rate of patients is about 20%.

The study of the relationship between obesity and breast cancer was first seen in the 1960s. Case-control studies by Dewaard et al. first proposed that women with breast cancer tend to have postmenopausal obesity and hypertension. It has also been found in animal experiments that the tumor growth rate of obese mice is significantly increased compared with the control group. Meta-analysis showed that the risk of breast cancer increased with the increase of BMI. The risk of breast cancer in patients with BMI>30kg/m2 was 1.3~2 times higher than that of normal people. Obesity can significantly increase the risk of breast cancer in women.

Summary of the invention

The main object of the present invention is to find new targets for anti-tumor to better prevent and/or treat tumors and control tumor proliferation and migration. The inventors found in the study that gene knockout technology or shRNA method is used to interfere with GPR1 gene expression, or to use specific antibodies. Interfering with the action of GPR1 receptor can effectively inhibit the proliferation and migration of human choriocarcinoma cells and breast cancer cells in vitro.

One aspect of the invention provides the use of a GPR1 gene or a GPR1 protein for screening for a pharmaceutical target for treating or preventing a tumor.

Another aspect of the invention provides the use of a GPR1 gene or GPR1 protein as a target for the prevention and/or treatment of tumors, or as a diagnostic target for tumors.

Another aspect of the present invention provides the use of an antagonist of the GPR1 gene or the GPR1 protein for the preparation of a medicament for preventing or treating a tumor.

In a specific embodiment of the present invention, an antagonist of the GPR1 gene or the GPR1 protein is disclosed. Use in the preparation of a medicament for preventing or treating tumor proliferation and migration.

Another aspect of the invention provides the use of an antagonist of the GPR1 gene or the GPR1 protein for preventing or treating a tumor.

The GPR1 gene or protein antagonist is selected from the group consisting of a GPR1 antibody, a GPR1 receptor antibody, a modified GPR1, a partial peptide of GPR1, siRNAs targeting GPR1 gene sequences, shRNAs, antisense molecules and DNase, or siRNAs, shRNAs, and antibodies. An expression vector for a sense molecule; for example, the shRNA is represented by SEQ ID No. 1.

In another specific experiment of the present invention, human chorionic cancer cell line JEG-3, which utilizes shRNA to interfere with the knockdown of GPR1 gene, was established by using shRNA sequence which specifically interferes with GPR1 gene expression, and the cell population proliferation experiment in vitro showed that it was compared with the control group. In contrast, the proliferation of human choriocarcinoma cells that knocked out the GPR1 gene was significantly inhibited. Further, it was found by scratch test that the migration ability of human choriocarcinoma cells that knocked down the GPR1 gene was significantly inhibited compared with the control group.

In a further aspect, the present invention provides an antagonist of a GPR1 gene or a GPR1 protein, which is an expression vector comprising a promoter and a nucleic acid insert operably linked to the above promoter, the nucleic acid insert, The insert is a shRNA, for example, the shRNA of SEQ ID No. 1.

In a specific experiment of the present invention, human breast cancer cells HCC1937 and MDA-MAB-231, which utilize shRNA interference to knock down the GPR1 gene, were constructed using shRNA sequences which specifically interfere with GPR1 gene expression, and in vitro cell population proliferation experiments showed that Compared with the control group, the proliferation of human breast cancer cells knocking down the GPR1 gene was significantly inhibited. Further, it was found by scratch test that the migration ability of human breast cancer cells that knocked down the GPR1 gene was significantly inhibited compared with the control group.

Accordingly, the present invention provides a method for preventing and/or treating a tumor comprising: targeting GPR1, reducing the expression level of GPR1 and/or antagonizing the action of GPR1 to prevent and/or treat a tumor. Specifically, it may be to reduce the expression level of GPR1 (knockout of the expression level of the GPR1 gene or the knockdown GPR1 gene) and/or antagonize the action of GPR1 with an antagonist against GPR1.

In the present invention, the GPR1 (G Protein-Coupled Receptor 1, G protein coupled receptor-1) is one of the receptors of the chemokine/adipeptide chemerin and belongs to the G protein coupled receptor family.

The GPR1 gene of the present invention is a gene known in the prior art and is the only sequence, GeneID of the NCBI: NM_146250. The specific sequence is shown in SEQ ID No. 4:

In the present invention, the terms "tumor" and "cancer" are used to mean the same meaning, which are all abnormal division and reproduction of cells in tissues. Tumors generally exhibit partial or complete lack of structural organization and functional coordination with normal tissues, and often form and differentiate tissue masses, which may be benign or malignant. The term malignant tumor or malignant tumor as used herein may also be a benign tumor. Often, unless adequate treatment is given, these malignant tumors can invade the tissue, can be transferred to different locations, and may recur after attempting to remove and cause death.

In the present invention, the tumor is preferably choriocarcinoma, placental villus, ovarian cancer, breast cancer, uterine cancer, cervical cancer, endometrial cancer, prostate cancer, liver cancer, pancreatic cancer, skin cancer, malignant melanin. Tumor, head and neck cancer, sarcoma, cholangiocarcinoma, bladder cancer, kidney cancer, colon cancer, testicular cancer, lung cancer, stomach cancer.

In the present invention, the treatment and prevention of tumor refers to prevention or treatment of tumor proliferation and migration.

In the present invention, the terms "antagonist" and "inhibitor" have the same meaning, and mean that the antibody against GPR1 is any agent which can lower the expression level of GPR1, antagonize the action of GPR1, and inhibit the binding of GPR1 to a ligand. This antagonist achieves this effect in a number of ways. One type of antagonist binds the GPR1 protein with sufficient affinity and specificity to neutralize the biological effects of the GPR1 protein. Such molecules include antibodies and antibody fragments. Another class of antagonists comprises fragments of proteins, mutant proteins or small organic molecules, ie, peptidomimetics, which will bind to a GPR1 or GPR1 binding partner, for example, a compound or small molecule polypeptide that blocks the binding of a chemokine to GPR1, thereby inhibiting The biological activity of GPR1. The GPR1 antagonist can be of any of these classes as long as it is a substance that inhibits the biological activity of GPR1. GPR1 Antagonists include GPR1 antibodies, GPR1 receptor antibodies, modified GPR1 and partial peptides of GPR1. Another class of GPR1 antagonists includes siRNAs, shRNAs, antisense molecules and DNases that are known in the art to target the GPR1 gene sequences disclosed herein. Such agents can be obtained according to the prior art by those skilled in the art, and can be any antagonist known in the art to reduce the expression level of GPR1 and/or antagonize the action of GPR1, or based on the molecular formula. An agent that has a function of lowering the expression level of GPR1 and/or antagonizing the action of GPR1 after modification and modification.

In a preferred embodiment of the invention, the antagonist against GPR1 is a small molecule antagonist, such as shRNA or the like. More preferably, the shRNA has the sequence set forth in SEQ ID No. 1.

Beneficial effect

The present invention demonstrates the relationship between the GPR1 gene and various tumors, and can be used as a target for the prevention and treatment of choriocarcinoma and breast cancer, and can effectively inhibit the chorion by using any technique to interfere with the expression of GPR1 gene or interfere with GPR1. Cancer and breast cancer cell proliferation and migration; provide a basis for disease prevention, diagnosis and treatment.

DRAWINGS

Figure 1: ShRNA GPR1 knockdown of GPR1 protein expression levels inhibits JEG3 choriocarcinoma cell proliferation.

Figure 2: ShRNA GPR1 knockdown of GPR1 protein expression levels inhibits migration of JEG3 choriocarcinoma cells.

Figure 3: ShRNA GPR1 knockdown of GPR1 protein expression levels inhibits breast cancer cell proliferation.

Figure 4: ShRNA GPR1 knockdown of GPR1 protein expression levels inhibits breast cancer cell migration.

Detailed ways

In order to more clearly understand the present invention, the present invention will be further described with reference to the following examples and the accompanying drawings. The examples are for illustrative purposes only and are not intended to limit the invention in any way. In the examples, each of the original reagent materials is commercially available, and the experimental methods not specifying the specific conditions are conventional methods and conventional conditions well known in the art, or in accordance with the conditions recommended by the instrument manufacturer.

Example 1: Interfering with the gene expression of GPR1, or antagonizing the action of GPR1, can effectively inhibit human Proliferation and migration of chorionic cancer cells.

This example demonstrates that the gene expression of GPR1 is interfered with, or the action of GPR1 is antagonized, and the proliferation and migration of human chorionic cancer cells can be effectively inhibited. Among them, a shRNA sequence that specifically interferes with the expression of the GPR1 gene was designed, and a human choriocarcinoma cell line that knocked down the GPR1 gene was interfered with by shRNA.

In vitro cell population proliferation experiments showed that the proliferation of human choriocarcinoma cells that knocked out the GPR1 gene was significantly inhibited compared with the control group. Further, it was found by scratch test that the migration ability of human choriocarcinoma cells that knocked down the GPR1 gene was significantly inhibited compared with the control group.

details as follows:

1. Establish a human choriocarcinoma cell line that knocks down the GPR1 gene.

The human GPR1 gene shRNA used in this experiment was pSM2c-Hu-shGPR1 (purchased from Open Biosystems), and the Small Hairpin sequence:

5'-TGCTGTTGACAGTGAGCGCACTCTCTGATTGTCATTATATTAGTGAAGCCACAGATGTAATATAATGACAATCAGAGAGTTTGCCTACTGCCTCGGA-3' (SEQ ID No. 1). The control group was pSM2c-Hu-scramble shRNA.

1 The vigorously growing choriocarcinoma cells: JEG3 were inoculated into 6-well plates at 5×10 ⁵ cells/well one day before transfection, and the cell fusion degree was 60% after the second day of culture;

2 Transfection was performed on the second day, and 3 μg of the plasmid was diluted with 200 μL of opti-MEM medium in a culture well of a 6-well plate, and 6 μL of Lipofectamine 2000 was diluted with 200 μL of opti-MEM medium, respectively. After light mixing, leave it at room temperature for 5 minutes;

3 gently mix the two tubes of diluted solution, and let stand at room temperature for 20 minutes;

4 The cells to be transfected were gently rinsed once with PBS, 600 μL of opti-MEM medium was added, and the mixed dilutions were gently added to the culture wells and placed in a carbon dioxide incubator for cultivation;

5 After 4 to 6 hours of culture, discard the medium used for transfection, and add 3 mL of complete medium to the well;

After 648 hours, the medium containing 1 μg/mL puromycin was selected for screening; after the cells no longer died, a chorionic cancer cell line stably expressing GPR1 shRNA was obtained.

7 Total RNA was extracted with TRIzol, 2 μg of RNA was quantified for reverse transcription (reverse transcription kit, purchased from Promega), and qPCR was performed using specific primer sequences.

The specific primer sequence used is the Hu-GPR1 primer sequence:

Fw 5'-AATGCCATCGTCATTTGGTT-3' (SEQ ID No. 2)

Rv 5'-CAACTGGGCAGTGAAGGAAT-3' (SEQ ID No. 3)

8 Compared with the choriocarcinoma cells transfected with pSM2c-Hu-scramble shRNA (ie, the choriocarcinoma cell line transfected with an empty vector, JEG3), the GPR1 gene expression level was only 38% ± 2.57%, as a knock The human choriocarcinoma cell line with the GPR1 gene is weakened and named: JEG3-shGPR1/LRH; subsequent experiments can be used.

2. In vitro proliferation and migration experiments of human choriocarcinoma cell line with GPR1 gene.

(1) MTT proliferation experiment:

1) The chorionic cancer cells JEG3-shGPR1/LRH were seeded at a density of 10,000 cells/well in a 96-well cell culture plate at a volume of 200 μL per well, and cultured for 0 hours, 12 hours, 24 hours, 48 hours, and 72 cells, respectively. Hours; a negative control was also established with a chorionic cancer cell line transfected with an empty vector.

2) 20 μL of MTT solution (5 mg/mL) was continuously placed in a carbon dioxide incubator for 4 hours at different detection time points;

3) Discard the supernatant, add 150 μL of DMSO (dimethyl sulfoxide), shake for 10 minutes, select the wavelength of 490 nm on the microplate reader for detection, and calculate the cell proliferation rate. The OD values measured on the microplate reader at different times of the culture were different, and the ratio of the OD value at different time points to the OD value at 0 hours was the cell proliferation rate.

The results of the test are shown in Figure 1. The results of the test showed that the cell proliferation rate of the human choriocarcinoma cell line JEG3-shGPR1/LRH, which weakened the GPR1 gene, was significantly decreased compared to the choriocarcinoma cell line transfected with an empty vector. At 24 hours, 48 hours and 72 hours, the cell proliferation rates of the two cells were significantly different, and the gap between the two increased gradually with time: at 24 hours, the control group proliferated 1.69 times, GPR1 weakened The group increased by 1.4 times; at 48 hours, the control group proliferated 3.3 times, and the GPR1 knocked group increased by 2.98 times; at 72 hours, the control group proliferated 5.6 times, and the GPR1 knocked group increased by 4.57 times. The test results show that when the GPR1 gene is inhibited, the corresponding choriocarcinoma cell proliferation rate also decreases, that is, by inhibiting the GPR1 gene or its expression, the proliferation of choriocarcinoma cells can be effectively inhibited.

(2) Scratch test

1) First use the marker pen on the back of the 6-well plate, evenly draw the horizontal line, about every 0.5 to 1cm, across the hole. Pass at least 5 lines per hole;

2) Add about 2×10 ⁶ JEG3 cells to the well: JEG3-shGPR1/LRH and untreated choriocarcinoma cells, overnight;

3) The next day, use the gun head to scratch the horizontal line behind the back;

4) Wash the cells 3 times with PBS, remove the delineated cells, and add serum-free or low serum medium;

5) The cells were cultured in a 37 ° C incubator, and photographed at 0 hours, 12 hours, and 24 hours after the culture.

The test results are shown in Figure 2. Among them, pSM2C shGPR1 was JEG3-shGPR1/LRH group, and pSM2C empty vector was a chorionic cancer cell line transfected with empty vector. Normalization statistics were performed at 12 and 24 hour migration distances. It can be seen that (right panel) the migration distance of GPR1-inhibited choriocarcinoma cells is significantly lower than that of choriocarcinoma cell lines transfected with empty vector, and There were significant differences at 12 hours and 24 hours: at 12 hours, the control group was 1 and the GPR1 knockdown group was 0.709; at 24 hours, the control group was 1 and the GPR1 knockdown group was 0.762. The test results indicate that inhibition of GRP1 in choriocarcinoma cells can effectively inhibit cell migration.

Example 2. Interfering with the gene expression of GPR1, or antagonizing the action of GPR1, can effectively inhibit the proliferation and migration of human breast cancer cells.

This example demonstrates that the gene expression of GPR1 is interfered with or antagonizes the action of GPR1, and can effectively inhibit the proliferation and migration of human breast cancer cells. Among them, a shRNA sequence that specifically interferes with GPR1 gene expression is designed, and a human breast cancer cell line that knocks down the GPR1 gene is interfered with by shRNA.

In vitro cell colony proliferation experiments showed that the proliferation of human breast cancer cells with weak GPR1 gene was significantly inhibited compared with the control group. As shown in Figure 4, HCC1937 cells grew 2.08 times at 24 hours, and the GPR1 genome proliferated. At 37 hours, the control group proliferated 2.85 times, and the weak GPR1 genome proliferated 1.98 times. At 72 hours, the control group proliferated 4.78 times, and the weak GPR1 genome proliferated 3.51 times. When MDA-MB-231 cells were used for 24 hours, the control group proliferated 3.01 times, and the weak GPR1 genome proliferated 1.97 times. At 48 hours, the control group proliferated 4.2 times, and the weak GPR1 genome proliferated 2.94 times. At 72 hours, the control group proliferated 6.77. Times, knockdown of the GPR1 genome proliferation was 5.23 times. Further, it was found by scratch test that the migration ability of human breast cancer cells with weak GPR1 gene was significantly inhibited compared with the control group: the migration distance of the control group was 0.26 cm at 24 hours in HCC1937 cells, and the migration distance of the GPR1 genome was weakened. It was 0.08 cm; at 48 hours, the migration distance of the control group was 0.46 cm, and the migration distance of the knockdown GPR1 genome was 0.21 cm. At 24 hours of MDA-MB-231 cells, the migration distance of the control group was 0.36 cm, and the migration distance of the weak GPR1 genome was 0.29 cm; At the hour, the migration distance of the control group increased by 0.33 cm, while the migration distance of the weak GPR1 genome increased by only 0.16 cm.

The specific test methods are as follows:

1. Establish a human breast cancer cell line that knocks down the GPR1 gene.

The human GPR1 gene shRNA used in this experiment was pSM2c-Hu-shGPR1 (purchased from Open Biosystems), and the Hairpin sequence: 5'-TGCTGTTGACAGTGAGCGCACTCTCTGATTGTCATTATATTAGTGAAGCCACAGATGTAATATAATGACAATCAGAGAGTTTGCCTACTGCCTCGGA-3' (SEQ ID No. 1). The control group was wild group cells that were not treated.

1 Select vigorous breast cancer cells: HCC1937 and MDA-MAB-231, inoculate 5×10 ⁵ cells/well in the 6-well plate one day before transfection, and culture until the second day, the cell fusion degree is 60%. ;

The specific primer sequence used is the Hu-GPR1 primer sequence:

Fw 5'-AATGCCATCGTCATTTGGTT-3' (SEQ ID No. 2)

Rv 5'-CAACTGGGCAGTGAAGGAAT-3' (SEQ ID No. 3)

8 Compared with transfected pSM2c-Hu-scramble shRNA, GPR1 gene expression level was only 37%±2.97% of its (wild group HCC1937), which was used as a weakening of human breast cancer with GPR1 gene. The cell line, and named: HCC1937-shGPR1/LRH and MDA-MAB-231-shGPR1/LRH; can be used in subsequent experiments.

2. In vitro proliferation and migration experiments of human breast cancer cell lines with GPR1 gene weakened.

(1) MTT proliferation experiment:

1) Breast cancer cells were seeded at a density of 2000 cells/well in a 96-well cell culture plate at a volume of 200 μL per well, and cultured for 12 hours, 24 hours, 48 hours, and 72 hours, respectively; a negative control was also established.

3) Discard the supernatant, add 150 μL of DMSO (dimethyl sulfoxide), shake for 10 minutes, select the wavelength of 490 nm on the microplate reader for detection, and calculate the cell proliferation rate. The test results are shown in Figure 3. It can be seen from Fig. 3 that for both breast cancer cells, the cell proliferation rate was significantly lower than that of the control group by the interference effect of shRNA. And this reduction in value-added rate can continue.

(2) Scratch test

2) Add about 5×10 ⁵ breast cancer cells to the well: overnight;

5) Incubate in a 37 ° C incubator, take samples after 24 h and 48 h.

The test results are shown in Figure 4. Among the two breast cancers, HCC1937 and MDA-MB-231, in the statistics of migration distance of 24 hours and 24-48 hours, it can be seen that the migration distance of GPR1 inhibited breast cancer cells is significantly lower than that of untreated. Breast cancer cells were significantly different at 24 hours and 48 hours. The migration distance of 24-48 hours is less than the migration distance of 24 hours, indicating that the migration ability of cells is weakened with time. The results showed that inhibition of GRP1 in breast cancer cells can effectively inhibit cell migration and become more and more obvious over time.

The present invention has been illustrated and described with respect to the specific embodiments thereof, and is not intended to limit the invention. More specifically, the present invention relates to GPR1 antagonist polypeptides, polynucleotides, antibodies, devices and kits disclosed herein and uses thereof, as well as controlling GPR1 levels, and various modifications can be made in accordance with the details, these modifications Program in the claims The scope of the invention and the scope of the claims are not deviated from the spirit of the invention.

Claims

Use of a GPR1 gene or GPR1 protein for screening for a drug target for treating or preventing a tumor.
A GPR1 gene or GPR1 protein is used as a target for the prevention and/or treatment of tumors, or as a target for tumor diagnosis.
An antagonist of an GPR1 gene or an antagonist of a GPR1 protein for use in the manufacture of a medicament for preventing or treating a tumor.
An antagonist of an GPR1 gene or an antagonist of a GPR1 protein for use in the manufacture of a medicament for preventing or treating tumor proliferation and migration.
An antagonist of the GPR1 gene or an antagonist of a GPR1 protein for preventing or treating a tumor.
An antagonist of the GPR1 gene or an antagonist of a GPR1 protein for preventing or treating tumor proliferation and migration.
According to the use of claims 3-6, the antagonist of the GPR1 gene or the antagonist of the GPR1 protein is selected from the group consisting of a GPR1 antibody, a GPR1 receptor antibody, a modified GPR1, a partial peptide of GPR1, siRNAs targeting the GPR1 gene sequence, shRNAs, and A molecule and a DNase, or an expression vector comprising siRNAs, shRNAs, and antisense molecules.
The antagonist of the GPR1 gene or the antagonist of the GPR1 protein is an expression vector comprising a promoter and a nucleic acid insert operably linked to the above promoter, according to the use of claims 3-6, The insert is a shRNA.
The use according to any one of claims 1 to 7, wherein the tumor is preferably choriocarcinoma, placental villus, ovarian cancer, breast cancer, uterine cancer, cervical cancer, endometrial cancer, prostate cancer, liver cancer, pancreatic cancer , skin cancer, malignant melanoma, head and neck cancer, sarcoma, cholangiocarcinoma, bladder cancer, kidney cancer, colon cancer, testicular cancer, lung cancer, stomach cancer.
A method for preventing and treating a tumor or preventing or treating tumor proliferation and migration, which comprises the steps of knocking out or knocking down the GPR1 gene, or downregulating the expression level of GPR1; preferably, this step is achieved by a GPR1 antagonist.