A method for detecting the ability and efficiency of tumors to produce primordial germ cells and its use

Abstract

The present invention belongs to the field of biological and medical testing technology, and relates to a method for detecting the ability and efficiency of a tumor to produce PGC-like cells, and specifically relates to a method for detecting the ability of a tumor to produce primordial germ cells (PGCs)-like tumor cells. The present invention uses tumor tissue or tumor tissue culture to detect the presence and / or formation efficiency of primordial germ cells (PGCs)-like tumor cells therein, and uses the detection of changes in the expression of genes related to the activation of PGC-like tumor cell formation in tumor tissue and tumor tissue culture to further predict the metastatic ability and metastasis risk of the tumor.

Description

Technical Field

[0001] The present invention belongs to the field of biological and medical testing technology, and relates to a method for detecting the ability and efficiency of a tumor to produce PGC-like cells, and specifically relates to a method for detecting the ability of a tumor to produce primordial germ cells (PGCs)-like tumor cells, which is further used to predict the metastatic ability of a tumor. Background Art

[0002] Studies have reported that tumor cells are heterogeneous, with only a small subset of cells playing a decisive role in tumor metastasis. Clinical practice has shown that circulating tumor cells (CTCs) are present in the body fluids of some tumor patients during surgical resection, but only approximately 10% of these patients will experience metastasis. Therefore, there is an urgent need in clinical practice to be able to promptly identify cell types that indicate a tumor's propensity to metastasize. Some researchers have speculated that cells with metastatic potential in tumors of different types and tissue origins may share certain common biological characteristics.

[0003] Studies have shown that there is a high degree of similarity between tumor metastasis and PGCs migration [1,2] ; They are highly similar in both behavior and mechanisms related to metastasis / migration ability, such as both require CXCR4-CXCL12 interaction, etc.

[0004] Based on the research foundation of existing technologies and the previous research findings of this research team [3-5] : "PGC-like tumor cells" exist among various types of tumor cells. The inventors of this application intend to provide a method for detecting the ability and efficiency of tumors to produce primordial germ cells (PGC-like tumor cells) and to activate the specific gene expression products formed by these cells as a means to predict tumor metastasis ability.

[0005] References related to the present invention include:

[0006] 1. Simpson, AJ, Caballero, OL, Jungbluth, A., Chen, YT & Old, LJ Cancer / testis antigens, gametogenesis and cancer. Nat. Rev. Cancer 5, 615–625 (2005).

[0007] Sun,

[0008] 3.Ma,Z et al.Spontaneous generation of germline characteristics inmouse fibrosarcoma cells.Sci.Rep.2,743(2012).

[0009] 4. Liu, C. et al. Activation of the germ-cell potential of human bonemarrowderived cells by a chemical carcinogen. Sci. Rep. 4, 5564 (2014).

[0010] 5. Liu, C et al. Abnormal gametogenesis induced by p53 deficiencypromotes tumor progression and drug resistance. Cell discovery 4, 54, doi:10.1038 / s41421-018-0054-x (2018).

[0011] 6. Aslakson CJ, Miller FR. Selective events in the metastatic process defined by analysis of the sequential dissemination of subpopulations of a mousemammary tumor. Cancer research; 52:1399-1405 (1992). Summary of the Invention

[0012] The purpose of the present invention is to provide a method and use for detecting the ability and efficiency of tumors to produce primordial germ cells based on the research foundation and current status of the existing technology, especially a method for detecting the ability and efficiency of tumors to produce primordial germ cells (PGC-like tumor cells) and the use of activating specific gene expression products formed by these cells as a predictor of tumor metastasis ability.

[0013] The present invention provides a method for detecting the ability and efficiency of a tumor to produce primordial germ cells, and predicting the ability of a tumor to metastasize by using the ability of a tumor to produce primordial germ cells (PGCs)-like tumor cells, which includes:

[0014] 1) Using patient-derived tumor tissue for primary and subculture, the ability to form PGC-like cells and the efficiency of formation can be used to predict whether the tumor has metastatic ability and the extent of its metastatic ability;

[0015] 2) Predicting tumor metastasis ability by detecting changes in the expression of a single gene or multiple genes associated with the activation of PGC-like cell formation in tumor tissue or tumor tissue culture.

[0016] In the present invention, the generation of PGC-like cells in primary and subcultured cells derived from patient's tumor tissue indicates the risk of tumor metastasis. The higher the content and generation efficiency of PGC-like cells, the higher the risk of tumor metastasis.

[0017] The present invention detects the presence and / or formation efficiency of primordial germ cells (PGCs)-like tumor cells in tumor tissues or tumor tissue cultures, and detects changes in the expression of genes related to the activation of PGC-like tumor cell formation in tumor tissues and tumor tissue cultures, which are further used to predict the risk of tumor metastasis.

[0018] The present invention conducted animal and cell experiments, and the results showed that the ability and efficiency of tumors to produce PGC-like cells play a decisive role in their metastatic ability.

[0019] In the present invention, different types of tumor cells are used: - / - PGC-like tumor cells are present in spontaneous thymic lymphoma of mice, subcutaneous tumors formed by mouse breast cancer cell 4T1, and liver micrometastases of subcutaneous tumors formed by mouse melanoma (e.g. Figure 1 shown).

[0020] The present invention experiments showed that compared with the original tumor, the liver micrometastasis had an enrichment of PGC-like tumor cells (such as Figure 2 a, b), in the implementation of the present invention, subcutaneous tumors formed by mouse breast cancer cells 4T1 were used as a model. The experimental results showed that the content of PGC-like tumor cells in the tumor cells appearing in the liver in the earliest stage of metastasis was as high as 99%; as the mouse tumor-bearing time prolonged, the content of PGC-like tumor cells in liver metastasis gradually decreased (as shown in FIG. Figure 2 c).

[0021] The present invention experiments show that PGC-like cells play a key role in liver metastasis. AP staining shows that although the content of PGC-like tumor cells (~4%) in 4T1 cells cultured in vitro is much lower than that of somatic cell-like tumor cells (~96%) (e.g. Figure 3 a), but the content of PGC-like tumor cells in suspended cells (a migration ability) was significantly higher than that of somatic cell-like tumor cells (as Figure 3 Although the content of PGC-like cells in 4T1 culture is much lower than that of somatic cell-like tumor cells, more than 99% of the metastatic cells in liver micrometastasis are PGC-like tumor cells after 4T1 cells are inoculated into the mouse tail vein (as shown in Figure 2). Figure 3 b);

[0022] In the embodiment of the present invention, p53 enriched in PGC-like tumor cells - / - Spontaneous thymic lymphoma cells and PGC-like tumor cells in mice contain very low levels of p53 - / - Mouse spontaneous myeloma was inoculated into nude mice via tail vein. The results of animal experiments showed that p53 - / - Spontaneous metastasis of thymic lymphoma cells in mice to the ovary, kidney, and liver and the relationship between p53 - / - Spontaneous myeloma in mice was significantly increased compared to that in controls (e.g. Figure 3 c, d), and the markers c-Kit and CXCR4, which are highly correlated with tumor metastasis, are mainly highly expressed in PGC-like cells (as shown in Figure 3 e), the results showed that PGC-like tumor cells play a key role in tumor metastasis.

[0023] The present invention experiments show that: inhibiting the formation of PGC-like tumor cells and thus inhibiting the formation and metastasis of tumors; after using Crispr-Cas9 technology to knock out the "embryonic / germ cell development-specific genes Oct4, Nanog, Prdm14, Vasa or DDX4", the ability of the knockout 4T1 cells to form "germ cell-like tumor cells" is extremely reduced compared with the control group cells, and thus their ability to form subcutaneous tumors is extremely reduced. In addition, their tumor metastasis ability is extremely reduced regardless of whether they are inoculated subcutaneously or by tail vein (such as Figure 4 The results confirmed that PGC-like tumor cells are highly consistent with natural PGCs in gene regulation and play a key role in tumor metastasis.

[0024] Based on literature reports: Different subtypes of mouse breast cancer cell lines 4T1, 168FARN, and 67NR from the same breast cancer have different metastatic abilities [6]Based on the results of previous studies, it is speculated that the three cell lines have different abilities to generate PGC-like tumor cells. The experimental results of the present invention show that 4T1 has the ability to generate AP+PGC-like tumor cells, 168FARN has only a trace amount of AP+PGC cells, and 67NR cells have almost no ability to generate AP+PGC-like cells. AP staining can well demonstrate the differences in the ability of the three cells to generate PGC-like cells.

[0025] Since the tumors produced by 67NR cells are benign and have no invasive growth, it is easy to judge from pathology that their metastatic ability is poor, while both 168FARN and 4T1 cells are invasive, and it is difficult to define the difference in metastatic ability between the two from a pathological perspective. The present invention uses RT-PCR to detect the expression of a series of genes related to germ cell development in 168FARN and 4T1 cell cultures. The RT-PCR results show that compared with 168FARN cells, 4T1 cells highly express Octamer-binding transcription factor 4 (Oct4), Nanog, Stellar / Stella, Nanos Homolog 3 (NANOS3), Fragilis (also known as mouse interferon-induced protein like gene-1, mil-1 / interferon-induced transmembrane protein 3, Ifitm3), Blimp1 (also as PRDM1), Sox17, Synaptonemal complex protein 3 (Scp3), Synaptonemal complex protein 1 (Scp1), PRDM14, and Deletedin azoospermia-like (DAZL), Mvh (also known as Vasa), AP; while low expression of Sox2 (such as Figure 5 As shown), further, the present invention uses the ratio of the RNA expression of a series of genes related to reproductive development to the RNA expression of the Sox2 gene to calculate the relative content of the two. Compared with 168FARN and 67NR, the ratio of the RNA expression of genes related to reproductive development to the RNA expression of the Sox2 gene in 4T1 cell culture showed a significant increase (p<0.001), as shown in FIG. Figure 5 shown);

[0026] Experiments have shown that the metastatic ability of tumors can be predicted by detecting single or combined detection of Oct4, Nanog, NANOS3, Fragilis (Ifitm3), Blimp1, Sox17, Stellar, Scp3, Scp1, PRDM14, DAZL, Mvh, AP, etc.; gene products include proteins and RNA.

[0027] The test of the present invention also shows that the ratio of one or more of Oct4, Nanog, Nanos3, Fragilis (Ifitm3), Blimp1, Sox17, Stellar, Scp3, Scp1, PRDM14, DAZL, Mvh (Vasa, DDX4), AP and the like to the gene product of Sox2 has higher specificity and sensitivity for predicting the metastatic ability of tumor cells; the gene product includes protein or RNA; for example, using Blimp1 / Sox2, Sox One or any combination of the 17 / Sox2, Stellar / Sox2, Fragilis / Sox2, PRDM14 / Sox2, Nanos3 / Sox2, Nanog / Sox2, AP / Sox2, Scp3 / Sox2, Scp1 / Sox2, DAZL / Sox2, and Mvh (Vasa, DDX4) / Sox2 ratios has higher sensitivity and specificity for predicting the metastatic ability of tumors; when Sox2 is also expressed, the larger the ratio, the stronger the metastatic ability.

[0028] In the present invention, the difference in the ratio of the expression of genes related to reproductive development to the expression of the Sox2 gene (including RNA and protein) is objectively present, and its calculation method includes all feasible formulas. The formula used to calculate the difference in RNA expression in this experiment is: 2 -[(目标基因QRT-PCR Ct值-内参QRT-PCR Ct值)-(Sox2 QRT-PCR Ct值-内参QRT-PCR Ct值) ]

[0029] In the present invention, the expression level difference of the above genes includes various forms such as comparison with the internal reference itself, comparison with the internal reference itself and then comparison with adjacent cancer tissue or normal tissue.

[0030] In the present invention, PGC-like tumor cells play a key role in tumor metastasis; therefore, the detection of the content and formation efficiency of PGC-like tumor cells in body fluids, tumor tissues or tumor cell cultures can be used as an effective method for predicting tumor metastasis.

[0031] The present invention relates to the detection of the above gene expression products including proteins and RNA.

[0032] The biological samples used for analysis in the present invention include but are not limited to body fluids, tumor tissues, and tumor cells in culture.

[0033] The body fluids involved in the present invention include blood, urine, pleural effusion, peritoneal effusion, cerebrospinal fluid, digestive fluid, semen, tumor puncture fluid, lymph fluid, etc.

[0034] The blood involved in the present invention includes whole blood, serum, plasma, separated nucleated cells, circulating tumor cells in the blood, etc.

[0035] The method of the present invention can be used for a wide spectrum of tumors, including all tumor types such as lung cancer, liver cancer, pancreatic cancer, liver cancer, gastric cancer, esophageal cancer, skin cancer, lymphoma, leukemia, kidney cancer, ovarian cancer, testicular cancer, prostate cancer, and nervous system tumors.

[0036] The detection of AP involved in the present invention includes immunohistochemistry, immunofluorescence and direct AP chemical staining to detect the presence of PGC-like tumor cells.

[0037] The RT-PCR involved in the present invention detects the RNA products of the above genes. After reverse transcription, the PCR involved includes: conventional PCR, fluorescent quantitative PCR, nested PCR, multiplex PCR, digital PCR, etc.

[0038] The present invention provides a method for detecting the ability and efficiency of a tumor to produce primordial germ cells. The method uses tumor tissue or tumor tissue culture to detect the presence and / or formation efficiency of primordial germ cells (PGCs)-like tumor cells therein, and uses the method to detect changes in the expression of genes related to the activation of PGC-like tumor cell formation in tumor tissue and tumor tissue culture, which is further used to predict the metastatic ability and metastasis risk of a tumor. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 Different types of tumor cells were shown. They metastasized from the primary tumor to the liver. PGC-like tumor cells were present in the earliest micrometastasis of the liver.

[0040] (a) Derived from p53 - / - PGC-like tumor cells appeared in liver micrometastases of spontaneous thymic lymphoma in mice. These cells were similar to primordial germ cells in morphology and gene expression; (b) p53 - / -Spontaneous thymic lymphoma in mice expresses Oct4, a marker of PGCs; (c) After subcutaneous inoculation of mouse breast cancer cells 4T1 and mouse melanoma B16-BL6 to form tumors, PGC-like tumor cells appear in liver micrometastases, which are similar to primordial germ cells in morphology and gene expression; (d) A series of PGCs markers are expressed in PGC-like tumor cells; Scale bar: 20 μm.

[0041] Figure 2 showed that PGC-like tumor cells were enriched in early liver metastases, among which

[0042] (a) and p53 - / - Compared with mouse spontaneous thymic lymphoma, mouse breast cancer cell 4T1 subcutaneous tumor, and mouse melanocytoma B16-BL6 subcutaneous tumor, PGC-like tumor cells are more abundant in early liver metastasis caused by these tumors; (b) HE staining shows: p53 - / - Macrometastases formed in the liver from spontaneous thymic lymphoma and mouse breast cancer cell 4T1 subcutaneous tumors; (c) Changes in PGC-like tumor cells in liver metastases formed from mouse breast cancer cell 4T1 subcutaneous tumors at different time points; the content of PGC-like tumor cells in the earliest liver micrometastases is as high as over 99%; Scale bar: 20 μm.

[0043] Figure 3 It was shown that PGC-like cells play a key role in liver metastasis, among which,

[0044] (a) AP staining characteristics of adherent and suspended PGC-like tumor cells in 4T1 culture. The bar graph shows the content of AP-positive PGC-like tumor cells in adherent and suspended cultures, respectively; (b) HE staining shows the results of Oct4 immunohistochemistry after 4T1 cells were inoculated via the tail vein. The bar graph shows the content of PGC-like tumor cells in inoculated tumor cells and early liver metastases; (c) p53 - / - Spontaneous thymic lymphoma in mice (high PGC content) and p53 - / - Spontaneous mouse sarcomas were inoculated into nude mice through the tail vein and metastasized to different organs; the sections were HE-stained for liver metastasis; (d) The bar graph shows: p53 - / - Spontaneous thymic lymphoma in mice (high PGC content) and p53 - / - The content of metastatic tumor cells in the liver after spontaneous sarcoma inoculation in mice via the tail vein; (e) Expression of the detected proteins in the orthotopic tumor and its liver metastasis tissue 3 weeks after subcutaneous inoculation; (f) Immunofluorescence analysis of the expression of the detected proteins in 4T1 and B16-BL6 cultures; **p < 0.01, Scale bar = 20 μm (a, b, g), 50 μm (d, f), 100 μm (c).

[0045] Figure 4 , inhibiting the formation of PGC cell-like tumor cells and inhibiting tumor metastasis, among which,

[0046] (a) AP staining in the control group and in 4T1 cells with germ cell-associated gene knockout; (b) The percentage of AP-positive cells in the control group and in 4T1 cells with germ cell-associated gene knockout; (c) Flow cytometry analysis: The percentage of Stellar+ cells in the control group and in 4T1 cells with germ cell-associated gene knockout; (d) Real-time fluorescence quantitative PCR analysis showed the expression of multiple PGC-associated genes in the control group and in 4T1 cells with germ cell-associated gene knockout; (e) HE staining showed the expression of PGC-associated genes in the control group and in 4T1 cells with germ cell-associated gene knockout. After the 4T1 cells with the knockout genome were inoculated into the subcutaneous tissues of mice to form tumors, the metastasis to the liver was observed in each group; (f) HE staining showed that after the control group and the 4T1 cells with the knockout genome were inoculated into the tail vein of mice, the metastasis to the liver was observed in each group; (g) The control group and the 4T1 cells with the knockout genome were inoculated into the subcutaneous tissues of mice to form tumors, and the weight of the subcutaneous tumors and the ability to metastasize to the liver were analyzed in each group; (h) The control group and the 4T1 cells with the knockout genome were inoculated into the tail vein 15 days later, and the ability of each group to metastasize to the liver was analyzed; Scale bar = 50 μm.

[0047] Figure 5 The ability to form PGC-like tumor cells plays a key role in the metastasis of breast cancer cell lines. (a) Differences in the metastatic ability of 4T1, 168FARN, and 67NR cells; (b) AP staining and morphological analysis of the content of PGC-like cells in 4T1, 168FARN, and 67NR; (c) RT-PCR analysis of the expression differences of a series of PGCs development-related genes in 4T1, 168FARN, and 67NR; (d) Differences in the ratios of mRNA expression of a series of PGCs development-related genes to their Sox2 mRNA expression in 4T1, 168FARN, and 67NR; ** P<0.001.

[0048] Figure 6 The ability to form PGC-like tumor cells plays a key role in the metastasis of cancerous bone marrow cells. (a) AP staining and morphological analysis of the content of PGC-like cells in cancerous bone marrow cells T2 and T7; (b) RT-PCR analysis showing differences in the expression of a series of PGC development-related genes in cancerous bone marrow cells T2 and T7; (c) The difference in the ratio of mRNA expression of a series of PGC development-related genes to Sox2 mRNA expression in cancerous bone marrow cells T2 and T7; (d) HE staining showing differences in the ability of cancerous bone marrow cells T2 and T7 to metastasize from subcutaneous in situ tumors to the liver.* P<0.05, ** P < 0.001. Scale bar = 50 μm.

[0049] Figure 7 The ability of PGC-like tumor cells to form plays a key role in the metastasis of human liver cancer cell line HL7702. (a) AP staining and morphological analysis of the content of PGC-like cells in HL7702-1 and HL7702-2; (b) RT-PCR showed the difference in the expression of a series of PGC development-related genes in HL7702-1 and HL7702-2; (c) The difference in the ratio of the mRNA expression of a series of PGC development-related genes to their Sox2 mRNA expression in HL7702-1 and HL7702-2; (d) HE staining showed the metastatic ability of HL7702-1 and HL7702-2 in the liver, respectively. * P<0.05, ** P < 0.001. Scale bar = 50 μm. DETAILED DESCRIPTION

[0050] Example 1

[0051] In mouse bone marrow tumor cells induced to become cancerous in mice, the content of PGC-like tumor cells plays a key role in their metastatic ability.

[0052] AP staining was used to analyze the differences in the content of PGC-like tumor cells in T2 and T7 cultures of cancerous bone marrow cells (e.g. Figure 6 shown);

[0053] RT-PCR showed that compared with T7 cultures of cancerous bone marrow cells, a series of germ cell development-related genes were highly expressed in T2 cultures of cancerous bone marrow cells; Sox2 was more highly expressed in T7 (e.g. Figure 6 shown);

[0054] The RNA expression levels of multiple germ cell development-related genes in T2 and T7 cultures were compared with the Sox2 RNA expression level using formula 2 -[(目标基因QRT-PCRCt值-内参QRT-PCRCt值)-(Sox2QRT-PCRCt值-内参QRT-PCRCt值)] The results showed that the ratio of a series of germ cell development-related genes to Sox2 in T2 culture was significantly higher than that in T7 (such as Figure 6 , as shown in Table 1);

[0055] The cancerous mouse bone marrow cell cultures of T2 and T7 were inoculated subcutaneously into nude mice. The animal experiment results showed that when the subcutaneous in situ tumor size (diameter 1.5 cm) was similar, the tumor-bearing mice inoculated with cancerous bone marrow cell T2 cells showed obvious metastasis; while the tumor-bearing mice inoculated with cancerous bone marrow cell T7 cells did not show any metastasis (such as Figure 6 shown);

[0056] The experimental results show that in the bone marrow tumor cells of mice induced to become cancerous, the content of PGC-like tumor cells is highly correlated with the tumor metastasis ability; AP staining and the change pattern of germ cell-specific related genes can be further used to predict the metastasis ability of mouse cancerous bone marrow cells.

[0057]

[0058]

[0059] Example 2

[0060] In the patient's liver cancer cell line HL7702, the ability to form PGC-like tumor cells plays a key role in its metastatic ability.

[0061] The liver cancer cell line HL7721 was obtained from the patient, and single clones were picked in 96-well plates. Cell subpopulations with different PGC-like production abilities were selected by AP staining, and two cell subpopulations, HL7721-1 (with a strong ability to produce PGC-like tumor cells) and HL7721-2 (with a weak ability to produce PGC-like tumor cells), were obtained (e.g. Figure 7 shown);

[0062] The expression of PGC formation-related genes in HL7721-1 and HL7721-2 cell lines was detected by QRT-PCR. The results showed that, compared with HL7721-2, a series of germ cell development-related genes except Sox2 were highly expressed in HL7721-1 culture; Sox2 was more highly expressed in HL7721-2 (e.g. Figure 7 shown);

[0063] The RNA expression levels of multiple germ cell development-related genes in HL7721-1 and HL7721-2 cultures were compared with the Sox2 RNA expression level using formula 2 -[(目标基因QRT-PCR Ct值-内参QRT-PCR Ct值)-(Sox2 QRT-PCR Ct值-内参QRT-PCR Ct值)] The results showed that the ratio of a series of germ cell development-related genes to Sox2 in HL7721-1 culture was significantly higher than that in HL7721-2 (such as Figure 7 , as shown in Table 2);

[0064] HL7721-1 and HL7721-2 cells were inoculated into 5 nude mice via the tail vein. The results of animal experiments showed that 3 weeks after inoculation, significant metastasis occurred in the nude mice inoculated with HL7721-1 cells, while no metastasis was observed in the nude mice inoculated with HL7721-2 cells (e.g. Figure 7 shown);

[0065] The experimental results show that: in human liver cancer cells, the content of PGC-like tumor cells is highly correlated with the tumor metastasis ability; AP staining and the change pattern of germ cell-specific related genes can be further used to predict the metastasis ability of human liver cancer.

[0066]

Claims

1. Use of a reagent for detecting A gene and Sox2 gene in the preparation of a kit for predicting tumor metastasis ability; wherein, 1) Using the ability and efficiency of PGC-like cell formation in tumor tissue to predict whether a tumor has metastatic ability and the extent of its metastatic ability; 2) Detect changes in the expression of a single A gene or multiple A genes associated with the activation of PGC-like cell formation in tumor tissue or tumor tissue culture to predict tumor metastasis ability; Wherein, the A gene is Oct4, Nanog, Stellar, Nanos3, Blimp1, PRDM14, IFITM3, Sox17, SCP1, SCP3 or AP; Wherein, the tumor is thymic lymphoma, breast cancer or melanoma.

2. The use according to claim 1, characterized in that The ratio of the gene to the Sox2 gene product is used as a prediction of tumor metastasis ability, including: Blimp1 / Sox2 ratio, Sox17 / Sox2 ratio, AP / Sox2 ratio, Nanos3 / Sox2 ratio, Stellar / Sox2 ratio, PRDM14 / Sox2, SCP1 / Sox2 ratio, Oct4 / SOX2, SCP3 / SOX2 protein or RNA expression ratio; the larger the ratio, the stronger the metastasis ability and the greater the credibility of the metastasis prediction; the protein or RNA detection method includes WB, immunohistochemistry, immunofluorescence, flow cytometry, and RT-PCR.

3. The use according to claim 1, characterized in that The changes in the expression of a single A gene or multiple A genes associated with the activation of PGC-like cell formation include changes in the single and combined expression levels of Blimp1 and SOX17 for predicting tumor metastasis ability, and also include combined use with other indicators to predict tumor metastasis ability.

4. The use according to claim 1 or 2, characterized in that The formula used to calculate the difference in RNA expression levels was: 2-[(target gene QRT-PCR Ct value-internal reference QRT-PCR Ct value)-(Sox2 QRT-PCR Ct value-internal reference QRT-PCR Ct value)].

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