MiRNA-128-3p of targeting Galectin-3 gene and use of miRNA-128-3p to resistance of pancreatic cancer
A pancreatic cancer, targeted technology, applied in the direction of DNA / RNA fragments, gene therapy, genetic engineering, etc., can solve the problems of cancer recurrence and specific killing of cancer cells by cancer stem cells
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Embodiment 1
[0123] Example 1 Galectin-3 expression and upstream miRNA prediction in pancreatic cancer cell lines
[0124] 1. Use Western Blot and qRT-PCR to confirm the expression of Galectin-3 in normal pancreatic ductal cells and pancreatic cancer cell lines. Western Blot and qRT-PCR were used to detect the expression of Galectin-3 in normal pancreatic ductal cells and pancreatic cancer cell lines.
[0125] Bioinformatic analysis of the relationship between Galectin-3 and pancreatic cancer using the GEPIA platform
[0126] It was found in the platform that by comparing the expression of Galectin-3 in 179 cases of pancreatic cancer specimens and 171 cases of normal specimens, the statistical analysis found that the expression of Galectin-3 in pancreatic cancer was significantly higher than that in normal people ( figure 1 A). Moreover, among pancreatic cancer patients, the survival time of the Galectin-3 low expression group (89 cases) was longer than that of the Galectin-3 high ex...
Embodiment 2
[0149] Example 2 Effect of miRNA-128-3p mimic on PANC-1 cells
[0150] The transfection method was used to transfect miRNA-128-3p mimic into PANC-1 cells, and Western Blot and qRT-PCR methods were used to detect the effect on the expression of Galectin-3 in PANC-1 cells; Methods, Transwell cell invasion assay and CCK-8 cell proliferation assay to verify the effect of miRNA-128-3p mimic on the migration, invasion and proliferation of pancreatic cancer PANC-1 cells.
[0151] 2.1 Transfection of miRNA-128-3p mimic into PANC-1 cells
[0152] Purchase miRNA-128-3p mimic, miRNA-128-3p mimic-NC control, miRNA-128-3p inhibitor for inhibiting miRNA-128-3p, miRNA-128-3p inhibitor-NC control from Shanghai Ruibo Biological Company, Among them, miRNA-128-3p mimic has red fluorescence, and the above four substances were transfected into PANC-1 cells using transfection reagent Lipofectamine2000, and the red fluorescence in PANC-1 cells was photographed with an inverted fluorescence micr...
Embodiment 3
[0176] Example 3 Collecting HUCMSCs and extracting EXO
[0177] Fresh umbilical cords of healthy newborns were collected, and HUCMSCs were obtained by tissue-adherence method; miRNA-128-3pmimic was transfected into HUCMSCs, and exosomes derived from HUCMSCs were extracted; HUCMSCs and exosomes derived from HUCMSCs were detected by qRT-PCR. The content of miRNA-128-3p mimic in the body.
[0178] 3.1 Collecting HUCMSCs and Observing Morphology
[0179] Fresh umbilical cords of healthy infants with a length of 10-12 cm and normal gestational age were collected. The puerpera had no infectious diseases such as hepatitis, syphilis, and AIDS. The consent of the puerpera and family members was obtained. Collected by tissue-adherent method
[0180] HUCMSCs were cultured for about (Figure 6A, 6B). Under the microscope, the cells were in the shape of spindle-shaped strips with large nucleoplasm (Figure 6C); when the cells were collected and subcultured, the cells were in a spiral s...
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