CN110172448A - A kind of synovial sarcoma cells system hSS-005R and its progeny cell system - Google Patents

Abstract

The present invention provides a synovial sarcoma cell line hSS-005R, which is preserved in China Center for Type Culture Collection, and the preservation number is CCTCC NO: C201911. The present invention also provides a progeny cell line of the aforementioned synovial sarcoma cell line hSS-005R. The invention establishes a new human synovial sarcoma cell line, which has stable properties and can be stably passed down for multiple times. The human synovial sarcoma cell line established by the present invention has the characteristics of high tumor formation rate, short incubation period and good uniformity under the premise of retaining the main clinical biological characteristics, enriches the synovial sarcoma cell bank, and can successfully prepare synovial sarcoma Animal models, the prepared animal models can be used for basic research and drug screening, providing powerful scientific research data for research based on the genetic background of the Chinese population, and also for the preclinical research of new drugs in vivo experiments for clinical anticancer drug sensitivity and Drug resistance testing provides new test materials.

Description

A synovial sarcoma cell line hSS-005R and its progeny cell lines

technical field

The invention belongs to the field of tumor cell research, in particular to a synovial sarcoma cell line hSS-005R and its progeny cell line.

Background technique

Synovial sarcoma (synovial sarcoma) is a highly invasive and highly malignant tumor. Synovial sarcoma accounts for 5-10% of all soft tissue sarcomas, and accounts for 10-20% of soft tissue tumors in adolescents and young adults. Between 1983 and 2012, the incidence of synovial sarcoma continued to increase from 0.906% to 1.548%. Synovial sarcoma occurs in the extremities, accounting for about 70% of all synovial sarcoma cases. The most common metastatic sites of synovial sarcoma are the lungs, followed by lymph nodes and bone marrow. Synovial sarcoma was originally named for its microstructural similarity to the surrounding synovial tissue when it is found in tissues near joints. However, some studies in recent years have shown that synovial sarcoma can appear in tissues lacking synovium such as the esophagus, lungs, and heart, which is contrary to this name. The exact cell origin of synovial sarcoma is still an unsolved mystery, and some scholars believe that it may come from mesenchymal stem cells. In addition, synovial sarcomas can display markers of differentiation between tumors of epithelial origin and tumors of mesenchymal origin, although synovial sarcomas have less histological similarity to tumors of both origins. At present, some scholars tend to define synovial sarcoma as a tumor of "uncertain differentiation". The World Health Organization defines synovial sarcoma as follows: a spindle cell neoplasm of mesenchymal origin with some degree of differentiation, including a glandular component. Synovial sarcoma can be histopathologically divided into: biphasic type, with varying degrees of epithelial and spindle cell components; (2) monophasic fibrous type, composed only of spindle cells; (3) monophasic type Epithelial type, composed mainly of epithelial cells, with only a few spindle cell populations present; (4) poorly differentiated type, which can resemble small round cell tumors with high-grade nuclear features, high mitotic activity and containing areas of necrosis, large cell/epithelial tumors or spindle cell tumors. Among the multi-group research data, the most common one reported is monophasic fibrous synovial sarcoma, which has a very high degree of malignancy and extremely rapid growth. Biphasic synovial sarcoma is the most characteristic type of synovial sarcoma, containing spindle cells and fibroblasts, and tumor cells formed by the combination of spindle cells and epithelioid cells migrate to each other. Some studies have suggested that the transition from monophasic to biphasic synovial sarcoma is not a jump, but a gradual transformation.

Most studies agree that: neoadjuvant chemotherapy or extensive tumor resection with radiotherapy is still the first choice for the treatment of synovial sarcoma. Even with standardized clinical treatment, the effective survival rate within 10 years is still less than 10%, only 8.9%. Therefore, improving the treatment plan of synovial sarcoma to improve the prognosis of patients has become a thorny problem.

At present, the main problem that limits the research progress of synovial sarcoma treatment is how to construct a reasonable and effective tumor model.

A cell line refers to a cell population that is propagated after the first successful passage of a primary cell culture. Especially the cells that survived the second death crisis after 40-50 divisions are called cell lines. Because the cells of the primary culture are generally passed on for about 10 generations, it is not easy to pass on, the growth of the cells will stagnate, and most of the cells will senescence and die. But there are a very small number of cells that can survive the "crisis" and continue to pass on. These surviving cells can generally pass on to 40--50 generations. This kind of passing cells is called a cell line. The genetic material of the cell line is unchanged. When the cells pass to 50 generations, there will be a "crisis" again, and they can no longer be passed on. However, the genetic material of some cells has changed and has the characteristics of cancer, which may be passed down indefinitely under culture conditions. This kind of passaged cells is called a cell line.

Cell lines capable of unlimited proliferation and stable passage can provide stable characteristics and sufficient raw materials for the research of synovial sarcoma, but currently the synovial sarcoma cell lines that can be purchased at home and abroad are very limited, among which the domestic cell bank can directly purchase There is only one synovial sarcoma cell line, SW-982. Moreover, the biological characteristics of existing synovial sarcoma cell lines are quite different from those of clinical synovial sarcoma. Therefore, for research related to synovial sarcoma cells, it is necessary to provide more new synovial sarcoma cell lines and their preparation methods and applications.

Contents of the invention

The technical problem to be solved by the present invention is to solve the low biological diversity of existing human synovial sarcoma cell lines, especially the low success rate of cell line establishment, and the biological characteristics of existing cell lines and clinical synovial sarcoma are quite different, etc. Insufficient, provide a new human synovial sarcoma cell line and its establishment method and application.

The present invention provides a synovial sarcoma cell line hSS-005R, which is preserved in China Center for Type Culture Collection, and the preservation number is CCTCC NO: C201911.

The present invention also provides a progeny cell line of the aforementioned synovial sarcoma cell line hSS-005R.

In a specific embodiment, the cell line highly expresses the c-myc oncogene.

In a specific embodiment, the SS18-SSX fusion gene is absent in said cell line.

The present invention also provides a method for constructing an in vivo mouse model of the above-mentioned cell line, comprising taking hSS-005R cells and inoculating mice subcutaneously on one side. After 1 to 3 weeks of inoculation, tumors begin to form and grow. The 50-day-old mouse is a mouse model for medical research.

In a specific embodiment, the mouse is an immunodeficiency mouse, preferably, the immunodeficiency mouse is a nude mouse or a SCID mouse.

The present invention also provides a preparation method of the synovial sarcoma cell line hSS-005R, the synovial sarcoma cell line is preserved in the China Center for Type Culture Collection, and the preservation number is CCTCC NO: C201911; the preparation method includes primary culture and subculture.

Those skilled in the art know that primary culture begins by directly growing a single layer of cells from a tissue block or dispersing the tissue into single cells by enzymatic or mechanical methods, and the culture before the first passage can be considered as primary culture. The biggest advantage of primary culture is that the tissue and cells have just been isolated, and the biological properties have not changed much, which can reflect the state in vivo to a certain extent. The definition of subculture is: after the monolayer cells formed by primary culture are confluent, they need to be separated and cultured, otherwise the cells will be depleted of nutrients due to insufficient space for generation or excessive cell density, which will affect the growth of cells. Often referred to as subculture or subculture. Usually, subculturing refers to expanding culture, that is, dividing a cell into two or three for culturing. Strictly speaking, however, transferring or transplanting cells from one flask to another, whether diluted or not, is called passaging or subculturing. Understandably, at any point in time, some cells are lost as they are inoculated from one bottle to another, so there must be some objective dilution of the cells.

In a specific embodiment, the primary culture is adherent culture or enzyme digestion culture. In the present invention, the enzymatic digestion method includes adding digestive enzymes to dissolve the tissue and release the cells. The enzymatic digestion method cultivation is a routine operation known to those skilled in the art.

In a specific embodiment, the primary culture is an adherent culture, and the method comprises cutting the surgically resected tumor specimen obtained from a patient with clinical human synovial sarcoma into thin pieces of 3-5 mm ³ , and cutting the thin pieces into thin pieces. The adherent cells are cultured on the bottom of the cell culture bottle. After 1-2 weeks of adherent culture, tumor cells gradually crawl out of the tumor tissue block, and then the primary cultured tumor cells are subcultured.

In a specific embodiment, the subculture includes discarding the culture solution in the culture bottle for primary culture, adding fresh trypsin cell digestion solution to the culture bottle, and stopping the digestion after the cells fall off, adding Shake or tap the fresh complete medium to break away from the bottle wall to form a cell suspension, resuspend by centrifugation and inoculate in a new culture bottle to continue culturing; each time when the cells grow to the entire culture bottle or grow to more than 80%, separate the bottles For subculture, each flask of cells is divided into 2 to 5 flasks during the subculture, and the offspring tumor cells are continued to be cultured in these new culture flasks.

The present invention also provides an application of the above-mentioned synovial sarcoma cell line hSS-005R in screening or preparing anti-synovial sarcoma cell drugs.

The present invention also provides an in vivo experimental method for screening candidate drugs for the treatment of synovial sarcoma. The in vivo experimental method includes the following steps: applying the test compound to a mammalian tumor model in which hSS-005R cells form tumors, and causing synovial sarcoma after administration. The test compound whose tumor volume decreases or disappears is the candidate compound for the treatment of synovial sarcoma.

In the present invention, the hSS-005R cells refer to cells from the synovial sarcoma cell line hSS-005R.

The present invention also provides an in vitro experimental method for screening candidate drugs for the treatment of synovial sarcoma. The in vitro experimental method includes the following steps: directly applying a test compound or a combination of different test compounds to hSS-005R cells at different concentrations, and according to different concentrations The inhibitory effect of the test compound or its combination on the proliferation of tumor cells was used to judge the anti-tumor ability of the test compound against synovial sarcoma.

In a specific embodiment, the external experiment method comprises the following steps: 1) seeding the hSS-005R cells in the wells of a 96-well cell culture plate at a density of 3000/well, and culturing for 24 hours; 2) The test compound was diluted into different concentrations and applied to hSS-005R cells. After 24 hours of drug action, the cell viability was measured to calculate the inhibitory ability of different concentrations of the compound to the cell proliferation to judge the anti-tumor ability of the test compound.

In a specific embodiment, the test compound includes cisplatin, doxorubicin and one or more of the c-myc inhibitors Kj-pyr-9 and 10058-F4.

The present invention also provides an application of the aforementioned synovial sarcoma cell line hSS-005R in preparing a synovial sarcoma cell model or an animal model, preferably, the animals in the animal model are immunodeficient mice.

The human synovial sarcoma cell line established by the present invention has the characteristics of high tumor formation rate, short incubation period, and good uniformity under the premise of retaining the main clinical biological characteristics, enriches the synovial sarcoma cell bank, and is based on the genetic background of the Chinese population. The research carried out provides powerful scientific research data, and also provides new test materials for the in vivo experiment of new drug preclinical research for the test of clinical antitumor drug sensitivity and drug resistance; and the human synovial sarcoma cell line has a clinical effect on synovial sarcoma The first-line drug, cisplatin resistance, is a good test material for studying the drug resistance mechanism of synovial sarcoma; it is also a set of excellent research materials for exploring new mechanisms of cisplatin resistance, which has high scientific research value and development significance.

The reagents and raw materials used in the present invention are all commercially available.

The beneficial effects of the present invention are at least:

1. The present invention establishes a new human synovial sarcoma cell line, which has stable properties and can be stably passed down for multiple times, and provides new experimental materials closer to clinical tumor biological characteristics for synovial sarcoma research.

2. The human synovial sarcoma cell line established by the present invention has the characteristics of high tumor formation rate, short incubation period, and good uniformity under the premise of retaining the main clinical biological characteristics, enriches the synovial sarcoma cell bank, and can successfully prepare synovial sarcoma cell lines. Membranous sarcoma animal model, the prepared animal model can be used for basic research and drug screening, providing powerful scientific research data for research based on the genetic background of the Chinese population, and also for preclinical research of new drugs in vivo experiments sensitive to clinical anticancer drugs Provide new test materials for testing of drug resistance and drug resistance.

3. The human synovial sarcoma cell line established by the present invention acquires drug resistance to synovial sarcoma clinical first-line chemotherapeutic drug—cisplatin, and it shows stable cisplatin resistance characteristics in vivo and in vitro experiments, which is resistant to cisplatin The exploration of the new mechanism is an excellent set of research materials with high scientific value and development significance, and it is a good test material for studying the drug resistance mechanism of synovial sarcoma.

4. The human synovial sarcoma cell line established by the present invention can be used to analyze the correlation between in vitro and in vivo drug sensitivity and drug resistance by comparing with the nude mouse in vivo passage parent tumor, and then can establish two correlations in vitro and in vivo. The drug screening platform is an ideal cell line for basic research and preclinical applications of human synovial sarcoma.

Preservation of Biological Material

The human synovial sarcoma cell line of the present invention was preserved in the China Center for Type Culture Collection (CCCTCC) (address: Wuhan University, Wuhan, China, zip code 430072) on January 15, 2019, and the culture name is: human synovial sarcoma cell hSS- 005R, the deposit number is: CCTCC NO: C201911.

Description of drawings

Figure 1 is the morphological observation of hSS-005R cells under an optical microscope. A. (100 times) B. (200 times).

Fig. 2 is the identification result of short segment repeat sequence (CSTR) of hSS-005R cells.

Figure 3 is flow cytometry analysis of hSS-005R cell cycle.

Figure 4 is the in vitro test of the inhibitory effect of multiple anticancer drugs on the proliferation of hSS-005R cells.

Figure 5 shows the tumorigenicity of hSS-005R cells and the growth curve of animal tumors in vivo. Wherein, A is the growth curve of the tumor in the nude mice, and B is the photo of the growth of the tumor in the nude mice.

Fig. 6 is the pathological tissue of primary tumor tissue and hSS-005R cells forming tumor in nude mice. Among them, A: primary tumor tissue; B: hSS-005R cells.

Detailed ways

The present invention is further illustrated below by means of examples, but the present invention is not limited to the scope of the examples. For the experimental methods that do not specify specific conditions in the following examples, select according to conventional methods and conditions, or according to the product instructions.

In the present invention, the hSS-005R cells refer to the cells preserved in the China Center for Type Culture Collection, and the preservation number is CCTCC NO: C201911, which come from the synovial sarcoma cell line hSS-005R.

Example 1

Example 1 is the preparation of the cell line hSS-005R of the present invention.

Primary culture method: Place fresh synovial sarcoma specimens resected during operation in DMEM high glucose + 10% FBS complete medium in a sterile state, and store them on ice. After rinsing with fresh PBS buffer (containing 1% double anti-penicillin, streptomycin), cut into thin slices, attach the cut tissue pieces to the bottom of the cell culture flask, add appropriate amount of DMEM high sugar +10 Complete culture with %FBS is based on 37°C cell culture incubator with 5% carbon dioxide and placed upright for 1 hour to make the tissue pieces stick firmly to the bottom of the bottle. The upright flask is then gently and slowly tipped over to submerge the adherent tissue pieces with complete medium. There is no need to change the medium, and the cells can usually be observed climbing out of the tissue block after 4-5 days. When the medium turns yellow, replace with a new medium, and when the cells are basically overgrown, carry out subculture according to the normal subculture method.

Subculture: Discard the culture medium in the culture bottle for primary culture, add fresh trypsin cell digestion solution to the culture bottle, stop the digestion after the cells fall off, add fresh complete medium to shake or tap to separate them The cell suspension is formed on the wall of the bottle, and it is centrifugally inoculated into a new culture bottle to continue the culture; each time the culture bottle is replaced, it is equivalent to subculture once; each time the cells grow to the entire culture bottle or grow to more than 80%, divide the bottle for subculture For culture, each flask of cells is divided into 2 to 5 flasks of the generation of tumor cells during subculture, and their progeny tumor cells are continued to be cultured in these new culture flasks.

In the present invention, the primary cells derived from tumor tissue were subcultured for 50 generations to obtain a cell line named hSS-005R, which was submitted for deposit with the deposit number CCTCC NO: C201911.

Example 2

Example 2 is the biological characteristics and application of hSS-005R cells.

The invention adopts DMEM high-sugar culture medium to cultivate and purify hSS-005R cells, so that the hSS-005R cells can grow and pass stably in vitro for a long time. When the cells are passed on for more than 30 generations, the cell traits are gradually stabilized, and relevant biology, genetics, and tissue source identification are carried out until the 50th generation has the same stable traits. The hSS-005R cells can form tumors in nude mice and have tumorigenicity. The corresponding relationship between the hSS-005R cells and the clinical synovial sarcoma tumor specimens derived from them, and the parental tumors passaged in nude mice can provide a new test for studying the correlation between in vitro, in vivo and clinical anticancer drug sensitivity and drug resistance Material. details as follows:

a. Morphological observation

Place the culture flask in which hSS-005R cells are cultured under an inverted microscope and observe under bright field. It can be seen that the main body of hSS-005R cells is like a paving stone, and there are a small amount of irregular cells, which show clonal growth and can appear when the density is high. Adhesive aggregation, the results are shown in Figure 1 (A100 times, B200 times).

b. Short segment repeat sequence (CSTR) identification

Short tandem repeat (short tandem repeat, STR), also known as microsatellite DNA, refers to a type of DNA sequence formed by tandem repetition on a chromosome with several base pairs as the core unit (2-6 base pairs). (The number of repetitions is more than 10-60, and the gene fragments are below 400 base pairs); the number of repetitions of each core unit will appear individual differences, thereby forming alleles with different fragment lengths. Therefore, the number of repetitions of a group of STR sequences is almost unique among different individuals, and it is an individual's genetic identity feature, and it is also the main method of cell biology to identify cell identity and origin.

Collect freshly cultured hSS-005R cells, use Tsingke’s Animal Genome Extraction Kit (Catalog No. TSP201-200) to extract the genomic DNA of the cells, perform PCR amplification with 5’ fluorescently labeled primers, sequence the obtained products, and analyze Including Amelogenin, THO1, TPOX, D13S317, vWA, D16S539, D5S818, CSF1PO, D7S820 and other STR site repeats, and the primer sequences of STR sites are shown in Table 1. The above sequence was queried and compared with the databases of ATCC, DSM2Z and other cell preservation banks, but the same genetic map was not returned. The STR sequence search in the American Type Culture Collection (CATCC) database did not find the same STR detection result. This can prove its uniqueness, and no cross-contamination with other cells occurred during the primary culture process (see Figure 2 for the test results).

It can be seen from Fig. 2 that the synovial sarcoma cell line hSS-005R provided by the present invention belongs to a brand-new human synovial sarcoma cell line.

Table 1: Primer sequences for STR loci

c. Cell cycle distribution

Collect about 10 ⁶ hSS-005R cells in a 1.5ml centrifuge tube, and discard the supernatant by centrifugation. The cell pellet was resuspended with 1 ml -20°C 75% ethanol, and fixed at room temperature for 1 hour. The supernatant was discarded by centrifugation, and 500 μl of PI staining solution was added. Mix well and incubate at room temperature for 30 minutes. Detect the number of cells per well and the total DNA content of each cell with a flow cytometer (the total DNA content of each cell is directly proportional to the total PI fluorescence intensity of the cell); and according to different cell cycles, the total DNA content of the cells Changes, the total number of cells in each cell cycle was calculated. The detected cycle distribution and the distribution ratio of each cycle are shown in Figure 3.

As known to those skilled in the art, the interphase of the cell cycle is divided into three phases, namely the early DNA synthesis phase (G1 phase), the DNA synthesis phase (S phase) and the late DNA synthesis phase (G2 phase). 1. The G1 phase (first gap) is a period from mitosis to before DNA replication, also known as the pre-synthesis period, during which RNA and ribosomes are mainly synthesized. This period is characterized by active material metabolism, rapid synthesis of RNA and protein, and a significant increase in cell volume. The main significance of this phase is to prepare material and energy for DNA replication in the next phase S phase. 2. S phase (synthesis) is the DNA synthesis phase, in which, in addition to synthesizing DNA, histones are also synthesized at the same time. The enzymes required for DNA replication are synthesized during this period. 3. The G2 phase (second gap) is the late stage of DNA synthesis and the preparation period for mitosis. During this period, DNA synthesis is terminated, and a large amount of RNA and protein are synthesized, including tubulin and maturation factors. The cell proliferation index refers to the ratio of the sum of S phase and G2 phase cells to the total number of cells, reflecting the proliferation speed of this group of cells. Increased S phase indicates active cell division.

It can be seen from FIG. 3 that the proliferative ability of the synovial sarcoma cell line hSS-005R cells in the present invention is active. The cells conform to the proliferation characteristics of tumor cells.

d. In vitro cytotoxicity test

In vitro determination of commonly used chemotherapeutic drugs for synovial sarcoma—cisplatin (Cislatin), doxorubicin (doxorubicin) and Kj-pyr-9 (c-myc inhibitor), 10058-F4 (c-myc inhibitor ) antiproliferative effect on hSS-005R cell line. The test cells were seeded in a 96-well plate at a density of 3000/well, and after 24 hours of administration of different concentrations of drugs and combined drugs, CCK8 was used to detect the cell viability at each drug concentration. Among them, the drug concentrations of the two drugs used in combination account for half each, and the total dosage is the same as that of other drugs with the same concentration. Among them, doxorubicin combined with Kj-pyr-9 (c-myc inhibitor) has the best drug effect, the results are shown in Figure 4.

e. Tumorigenicity of cells

Culture and collect hSS-005R cells in vitro, subcutaneously inoculate nude mice (purchased from Hunan Slack Jingda Experimental Animal Co., Ltd.), each animal was inoculated with 0.2ml, containing 1.0×107 cells, and ⁶ mice were inoculated on one side Inoculation, N=6, animal body weight and tumor size were monitored weekly. About 2 weeks after inoculation, tumors began to form and grow, and the tumor formation rate was basically 100%. The tumor growth curve was drawn, wherein tumor volume=(length×width×width)÷2; the results are shown in FIG. 5 . It can be seen from Figure 5 that hSS-005R cells can form tumors in immunodeficient mice and grow uniformly and rapidly.

f. After hSS-005R cell tumors were subcutaneously formed in mice, the tumors were removed, fixed and embedded in paraffin, and sections were prepared and stained with HE. Figure 6A is the tumor mass of the original tissue, and Figure 6B is the tumor mass of the hSS-005R cell mouse. The pathological diagnosis result is synovial sarcoma, and the tissue morphology is highly close to the tumor mass of the original tissue. The results are shown in Figure 6. This shows that the cell line hSS-005R of the present invention can well reflect the situation of the original tumor tissue.

g. Whole-exon gene fusion analysis

From the results of gene fusion, hSS-005R cells do not contain the phenomenon of SSX gene fusion, and the number of readable (read) supporting gene fusion is very small, which can basically be judged as background noise. Therefore, the tumor model of synovial sarcoma in this case is a clinically rare synovial sarcoma with negative expression of SS18-SSX. The results are shown in Table 2.

Table 2: Fusion Gene Results

Those skilled in the art know that most synovial sarcoma has a translocation of t(X,18; P11p,q11), which produces the most important specific fusion gene SS18-SSX in synovial sarcoma, which is translated into A fusion protein SS18-SSX was formed. And this chromosomal translocation mutation has always existed in the whole process of synovial sarcoma cell development. Therefore, some scholars believe that the fusion gene generated by chromosomal translocation in synovial sarcoma, SS18-SSX, may serve as the proto-oncogene of synovial sarcoma, driving the occurrence and development of tumors. Given that the SS18-SSX fusion gene exists in the vast majority of synovial sarcoma and is highly specific, this provides an important diagnostic idea for the specific diagnosis of synovial sarcoma: detection of the fusion gene SS18-SSX by molecular detection means The presence of synovial sarcoma is diagnosed with high specificity. Although for synovial sarcoma, the SS18-SSX fusion gene is present in more than 95% of synovial sarcoma patients, less than 5% of synovial sarcoma does not contain the SS18-SSX fusion gene.

In other words, the synovial sarcoma cell line hSS-005R provided by the present invention is a rare type, which may have a relatively higher degree of malignancy, and there may be fewer types of drugs that can treat this type, and related research will also be conducted accordingly. less.

h.MYC copy number changes

Chromosome 8 in hSS-005R cells had a large number of copy number amplifications and a small number of copy number deletions. The underlined interval in Table 3 covers the MYC gene, its CNV copy number changes up to 7, ProbCall is 0.999999998, and the accuracy is high. The results are shown in Table 3.

Table 3: MYC copy number changes

Those skilled in the art know that the c-myc oncogene is related to the occurrence and development of various tumors. The cell line of the present invention has high expression of the c-myc oncogene through gene exon sequencing expression. This sequencing demonstrated the cell immortality properties of this synovial sarcoma cell line.

The above content is a further detailed description of the present invention in combination with specific preferred embodiments, and it cannot be assumed that the specific implementation of the present invention is limited to these descriptions. For those of ordinary skill in the technical field of the present invention, without departing from the concept of the present invention, some simple deductions and substitutions can be made, which should be regarded as belonging to the protection scope of the present invention.

Claims (6)

1. a kind of synovial sarcoma cells system hSS-005R, is deposited in China typical culture collection center, deposit number is CCTCC NO:C201911.

2. the progeny cell of synovial sarcoma cells system hSS-005R as described in claim 1 a kind of is.

3. cell line according to claim 1 or claim 2, which is characterized in that the cell line c-myc oncogene height expression.

4. cell line according to claim 1 or claim 2, which is characterized in that there is no SS18-SSX to merge base in the cell line Cause.

5. a kind of construction method of the mouse in vivo models of the cell line as described in any one of Claims 1 to 4, including take Single side Mice Inoculated under hSS-005R cell skin, after inoculation 1~3 week, tumour is initially formed and is grown, 30~50 days after inoculation Mouse is medical research mouse model.

6. construction method according to claim 5, which is characterized in that the mouse is immunodeficient mouse, preferably described Immunodeficient mouse is nude mice or SCID mouse.