Tumor cell sugar uptake inhibitor and application thereof
A tumor cell and anti-tumor cell technology, applied in the field of genetic medicine, can solve the problems of unclear long-chain non-coding RNA related mechanism, cell cycle arrest, etc.
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Embodiment 1
[0030] Embodiment one 18 Fluoro-deoxyglucose uptake rate detection
[0031] Cloning of long-chain non-coding RNA (sequence as SEQ ID NO: 1) loaded on lentiviral vectors, directly infecting K562 cells, constructing stable cell lines with high expression of long-chain non-coding RNA, and empty virus-infected cells as controls; two cell lines The expression results see figure 1 .
[0032] The radiation dose (F) and cell radiation dose (B) of the culture supernatant of the above two kinds of cells were detected respectively by the γ-ray measurement detector, and the calculation was carried out according to the formula 18 Fluoro-deoxyglucose uptake rate (%)=B / (B+F)×100%. The leukemia K562 stable cell line with high expression of long non-coding RNA, compared with the non-high expression group, 18 Fluorine-deoxyglucose uptake rate decreased significantly (see appendix figure 2 ).
Embodiment 2
[0033] Example 2 Detection of GLUT10 gene level
[0034] According to the method of Example 1, the leukemia K562 stable cell lines with high expression and non-high expression of long-chain non-coding RNA were constructed, and after low-glucose induction treatment, the cells were collected, RNA was extracted by Trizol one-step method, and GLUT10 gene was detected by qRT-PCR method Level. Compared with the blank group, the level of GLUT10 gene decreased in the long non-coding RNA high expression group (see attached image 3 ).
Embodiment 3
[0035] Example 3 Detection of GLUT10 protein level
[0036] According to the method of Example 1, the leukemia K562 stable cell line with high expression and non-high expression of long-chain non-coding RNA was constructed, and after low-glucose induction treatment, cultured for 24 hours, collected cells, treated with PI and Western protein lysate, and extracted protein , using Western Blot method to detect GLUT10 protein level. In the leukemia cell line K562, the GLUT10 gene level decreased in the long non-coding RNA high expression group compared with the control group (see appendix Figure 4 ).
[0037] The invention packs long-chain non-coding RNA into a lentiviral vector to construct a sugar uptake inhibitor, detects the level of sugar uptake, and detects the GLUT10 gene and protein levels of tumor cells. By inhibiting its expression through gene drug technology, it can inhibit the sugar uptake of tumor cells and reverse the energy source advantage of tumor cells, there...
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