Functions and applications of beta 1 protein in hepatoma cells
A liver cancer cell and protein technology, applied in the field of biomedicine, can solve the problems of enhanced cell adhesion and reduced metastasis
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Embodiment 1
[0035] Example 1: Expression of protein RNA and protein in liver cancer cells Hep3B and HepG2, and their correlation with cell invasion ability
[0036] This example describes the expression of β1 protein in liver cancer cells and its correlation with cell invasion ability. The expression of β1 protein is high, and the ability of cell invasion is weak; the expression of β1 protein is low, and the ability of cell invasion is strong.
[0037] This example describes a method for detecting the expression of β1 protein in liver cancer by using techniques such as PCR or Western Blotting to detect the mRNA or protein level of β1 protein.
[0038] Human liver cancer cell lines Hep3B and HepG2 were inoculated in RPMI1640 medium containing 10% newborn bovine serum, and normal liver cell line HL-7702 was inoculated in RPMI1640 medium containing 10% fetal bovine serum. 2 cultured in a 5% carbon dioxide incubator.
[0039] When the cell confluency in the cell flask reaches 90%-100%, disc...
Embodiment 2
[0043] Example 2: The method of RNA interference was used to interfere with the expression of β1 protein, the expression level was down-regulated after the interference, and the migration and invasion ability of HepG2 cells were significantly increased
[0044] This example describes how the expression level of β1 protein affects the ability of cell invasion and migration. The expression of β1 protein was down-regulated, and the ability of cell migration and invasion increased.
[0045] Culture HepG2 cells, when the cell coverage rate is 30%~50%, according to the operation instructions of the Lipofectamine 2000 transfection kit, transfect the cells with interfering RNA to interfere with the expression of β1, use the random sequence as a negative control, and detect β1 according to the method in Example 1 mRNA, protein level expression and cell migration and invasion ability.
[0046] Experimental results such as figure 2 As shown, the expression of β1 mRNA and protein in He...
Embodiment 3
[0047] Example 3: After transfection of β1 protein gene (nucleic acid) in Hep3B cells, the migration and invasion abilities of the cells were significantly reduced
[0048] This example describes nucleic acid substances as examples, targeting the β1 protein to reduce the ability of cell migration and invasion.
[0049] Culture Hep3B cells, when the cell coverage (confluence) is 80%-90%, follow the instructions of the Lipofectamine 2000 transfection kit, transfect the cells with pcDNA3.0-hSCN1B, and use the empty plasmid as a negative control, according to Example 1 Methods The expression of β1 protein and the ability of cell migration and invasion were detected.
[0050] The experimental results showed that the β1 gene was expressed in Hep3B cells after transfection ( Figure 4 ), and the migration and invasion ability of cells transfected with β1 in Hep3B cells decreased (*P Figure 5 ).
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