Application of regulating EIF4A3 expression to regulate and control proliferation capacity of hepatoma carcinoma cells
A liver cancer cell, proliferation ability technology, applied in the field of bioinformatics, can solve problems such as unclear role
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Embodiment 1
[0069] like figure 1 As shown, the present invention provides a flowchart of the molecular mechanism and application of regulating the expression of EIF4A3 to affect the proliferation of liver cancer cells. Four types of liver cancer data were obtained from the TCGA database. After differential expression analysis, genes related to EIF4A3 expression were identified, and modules related to liver cancer were explored through weighted gene co-expression network analysis. After functional enrichment analysis, EIF4A3 was constructed to participate in liver cancer cells. An integrated regulatory network for proliferation.
[0070] like figure 1 As shown, the present invention provides a method for constructing a comprehensive regulatory network that regulates the expression of EIF4A3 to affect the proliferation of liver cancer cells, comprising the following steps:
[0071] (1) Data processing:
[0072] Download liver cancer data from the Cancer Genome Atlas (TCGA) database to obta...
Embodiment 2
[0091] (1) Construction of a cell line that knocks out EIF4A3:
[0092] Prepare recombinant EIF4A3-shRNA lentivirus and negative control (NC) lentivirus; infect LM3 cells with a lentiviral vector (EIF4A3-KD) that knocks out the EIF4A3 gene, and obtain stably infected knockout EIF4A3 cell lines and control cell lines. like Figure 5 A shows the transfection efficiency after knocking out EIF4A3, indicating that the liver cancer cell line LM3 has indeed knocked out EIF4A3. The specific process is as follows:
[0093] qRT-PCR was used for detection, and the sequencing results were compared with the sequence of EIF4A3. Log in to the Public TRCPortal website, search for EIF4A3 and select the verified and high-scoring shRNA interference sequence 5′-GGATGAAGCTGATGAAATG-3′ of EIF4A3 from the results, and compare it by BLAST homology. The synthetic shRNA template (nucleotide sequence as shown in SED ID NO: 1, specifically GGATGAAGCTGATGAAATG; the specific synthesis process is aimed a...
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