Application of RNA helicase DHX33 inhibitor in preparation of medicine for treating liver cancer
An RNA helicase, a technology for the treatment of liver cancer, applied in the field of biomedicine, can solve the problems of lack of targets and inhibitors for liver cancer, difficulties, and the increase of human liver cancer research and development targets, and achieve important medical development value and the effect of treating liver cancer
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[0054] The present invention also provides a method for preparing the lentivirus, comprising the following steps:
[0055] Use Lipofectamine 2000 (Life Technologies) liposome introduction method to transfect the plasmid mixture in 293T cells: the specific operation method is to use a cell culture dish with a diameter of 10 cm for transfection, and when the 293T cells grow to 90% confluence, the The following plasmids were mixed, including pLKO.1-shRNA (that is, the above lentiviral plasmid), pCMV-VSV-G, pCMV-dR8.2 dvpr, and the ratio of plasmid mixing was 9:8:1, so that the total DNA amount reached each Petri dish 12μg. Among them, the vectors pLKO.1, pCMV-VSV-G, pCMV-dR8.2 dvpr were purchased from Biovector Inc.
[0056] After 16-18h, it was changed to medium containing antibiotics (penicillin and streptomycin), and after another 24h or 48h, the cell culture medium was collected with a sterile pipette, centrifuged at 1000rpm for two minutes, and the virus was repacked in 5mL...
Embodiment 1
[0150] Example 1 DHX33 protein plays an important role in hepatoma cells, and gene silencing of DHX33 significantly inhibits the growth of human hepatoma cells in vitro and in vivo
[0151] 1. Immunohistochemical analysis of the expression of DHX33 protein in human hepatocellular carcinoma tissues
[0152] Microarrays of paraffin tissue sections of human liver cancer tissues were purchased from US BIOMAX Company, including 40 different liver cancer tissues in total, and three normal liver tissues were used as normal tissue controls. Paraffin-embedded tissue chips were first incubated in a 60°C oven for 30 minutes, then rapidly deparaffinized in xylene and gradually decreased at a range of ethanol concentrations (100%, 95%, 70%, 50%, and 25%). The solution was gradually hydrated (5 minutes of gentle shaking each time, and the treatment of each concentration of ethanol solution was repeated once), and the hydration was continued in distilled water for 10 minutes. Antigens were ...
Embodiment 2
[0162] Example 2 DHX33 regulates de novo cholesterol production in human hepatoma cells by regulating the transcription of various genes in the mevalonate pathway
[0163] Cell growth is inseparable from the synthesis of cell membranes, especially in cancer cells, the efficiency of membrane formation is significantly improved. Studies have shown that in liver cancer cells, the mevalonate pathway is abnormally active. The mevalonate pathway is an important biosynthetic route to generate cholesterol. It not only provides an important membrane component-cholesterol, but also can synthesize a variety of intermediates such as Endoyl lipid molecules, etc., promote the activation of various oncogenes, such as the farendylation of RAS protein, a lipid coupling reaction can activate RAS in the inner membrane. Therefore, the mevalonate pathway is an important factor in promoting the occurrence and development of cancer. If the important genes of this pathway are inhibited, studies have ...
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