Tumor stem cell detection method and application thereof
A technology of tumor cells and detection methods, applied in the determination/testing of microorganisms, biochemical equipment and methods, measuring devices, etc.
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Embodiment 1
[0065] p53 - / - PGC-like tumor cells play a key role in metastasis in spontaneous thymic lymphoma in mice
[0066] Mechanical method (two slides) p53 - / - Tumor cells in mouse spontaneous thymic lymphoma tissue were released into PBS;
[0067] Add Stellar antibody to tumor-containing cells (1*10 6 / 100ul) of PBS solution, incubated on ice for 1 hour;
[0068] Wash three times with PBS;
[0069] Add fluorescently labeled secondary antibody such as FITC, and incubate on ice for 1 hour;
[0070] Wash three times with PBS;
[0071] Use the isotype control as a blank, and sort out Stellar by flow cytometry high Cells and Stellar low cell;
[0072] Will Stellar high Cells and Stellar low The cells were inoculated into nude mice through the tail vein; 3 weeks after the tail vein inoculation, the mice were sacrificed;
[0073] Measure the metastasis, take pictures, and use HE staining to show the tumor cells in the metastatic tissue;
[0074] Animal experiment results show: ...
Embodiment 2
[0077] In mouse breast cancer cell line 4T1 / 168FARN, the content of PGC-like tumor cells plays a key role in its metastatic ability
[0078] The mouse breast cancer cell lines 4T1 and 168FARN cells derived from the same tumor, literature reports that 4T1 has the ability to transfer but 168FARN does not have the ability to transfer; in order to verify the positive correlation between PGC-like tumor cells and the ability to transfer, the present invention first uses AP staining to show : Compared with 168FARN cultures, PGC-like tumor cells were extremely elevated in 4T1 cultures; almost no AP was detected in 168FARN cells + PGC-like tumor cells (P Image 6 a);
[0079] RT-PCR results showed that compared with 168FARN culture, PGCs-related genes were highly expressed in 4T1 culture, suggesting that the content of PGC-like tumor cells in 4T1 culture was significantly higher than that in 168FARN culture (P Image 6 b);
[0080] Immunohistochemical detection showed that PGCs-related ...
Embodiment 3
[0086]The content of PGC-like tumor cells plays a critical role in their metastatic ability in mouse bone marrow tumor cells induced by carcinogenesis
[0087] The difference in the content of PGC-like tumor cells in T2 and T7 cultures of cancerous bone marrow cells was analyzed by AP staining (P Figure 7 a);
[0088] RT-PCR showed that compared with the T2 culture of cancerous bone marrow cells, the expression of PGC-related genes in T2 culture of cancerous bone marrow cells and the expression of PGC-related genes in T7 were significantly increased (P Figure 7 b);
[0089] The cultures of cancerous bone marrow cells T2 and T7 were respectively inoculated subcutaneously in nude mice. The animal experiment results showed that compared with the culture of cancerous bone marrow cells T2, the tumor ability of the cultures of cancerous bone marrow cells T2 was significantly increased (P Figure 7 c);
[0090] Animal experiments showed that when the size of subcutaneous orthotopic tu...
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