Whole Transcriptome Analysis Of Sorafenib On Gene Expression Of Human Hepatoma Cells(HepG2)

Hepatocellular carcinoma(HCC)is a very common malignant tumor in China,with a high incidence and low survival rate.Sorafenib is a targeted chemotherapy drug,administered orally,for the treatment of patients with advanced liver cancer.Clinical data show that patients with advanced HCC taking sorafenib have a much longer overall survival than conservative palliative care.Treatment,but there are some limitations in the treatment process.Therefore,an in-depth understanding of the mechanism of action of sorafenib on liver cancer is of great significance in improving the therapeutic effect of sorafenib.Objective: The objective of this study was to explore the effects of sorafenib on the expression of human hepatocellular carcinoma(HepG2)mRNA,miRNA,LncRNA and CircRNA based on high-throughput sequencing technology,and to initially excavate the role of sorafenib from the level of whole transcriptome.Closely related response genes and signaling pathways to explore the molecular mechanism of sorafenib treatment of hepatocellular carcinoma,and provide scientific basis for elucidating the mechanism of action of sorafenib.Methods: 1.CCK-8 kit was used to detect the effect of sorafenib on proliferation of hepatocellular carcinoma cells;Flow cytometry was used to detect the effect of sorafenib on apoptosis and cell cycle of hepatoma cells;2.Whole transcriptome sequencing was used separately.Sorafenib stimulated hepatocellular carcinoma cells for 24h(HepG2-24h)and 48h(HepG2-48h),extracted control group cells(HepG2-0h)and drug-treated hepatoma cells Hep-G2-24 h and HepG2-48 h total RNA.Sample RNA was detected in quality,unrelated RNA was removed,followed by reverse transcription,PCR amplification and purification,and finally on-machine high-throughput sequencing,systematic analysis of the sequencing results.Results: 1.Sorafenib inhibited the proliferation of human hepatocellular carcinoma cell HepG2,promoted the apoptosis of human hepatocellular carcinoma cell HepG2,mainly in the late stage of apoptosis;sorafenib could inhibit the cell cycle of human hepatoma cell HepG2 in G1 phase;A total of 631 differentially expressed genes were detected in this study: HepG2-24 h group had a total of 631 differentially expressed genes,of which248 were up-regulated and 383 were down-regulated;1069 genes were differentially expressed in Hep-G2-48 h,of which,There were 457 increased expression levels and612 decreased expression levels;only 380 non-coding RNAs(miRNA: 172,CircRNA:155,lncRNA: 53)and 468 encoding RNAs were differentially expressed in the HepG2-24 h group.Only 547 non-coding RNAs(miRNA: 187,Circ RNA: 239,lncRNA: 121)differentially expressed in HepG2-48 h and 797 encoding RNAs;differentially expressed in HepG2-24 h and HepG2-48 h A total of 547 non-coding RNAs(miRNA: 109,CircRNA: 47,and lncRNA: 33)were used for GO analysis of differentially expressed genes shared by the HepG2-24 h and HepG2-48 h groups of cells: most genes were partially enriched for molecular function.Binding site(288)including protein binding 220),catalytic activity(121),ion binding(89),etc.;cell components are mainly concentrated in the cell(320),including cell part(319),intracellular(269),intracellular part(263);biological process Some of them mainly deal with single biological processes(305),single biological cellular processes(275),cellular processes(310),etc.;KEGG signaling pathway analysis found that genes with the same differential expression in HepG2-24 h and HepG2-48 h groups are mainly enriched In: Metabolic Pathways(52),Metabolism of xenobiotics by cytochrome P450(11),Drug metabolism-cytochrome P450(10),HTLV-I infection(10),Chemical carcinogenesis(10),MAPK signaling pathway(10),etc..Conclusion:1.Sorafenib can inhibit the proliferation of HepG2 cells,promote apoptosis,and arrest cell cycle in G1 phase.2.Sorafenib can significantly influence the expression of human hepatocellular carcinoma(HepG2)mRNA,and influence the biological behaviors of HepG2 cells such as cell proliferation,apoptosis and cell cycle by regulating the expression of tumor-associated genes.3.Sorafenib induced abnormal expression of miRNAs.These abnormally expressed miRNAs regulate theproliferation,apoptosis,migration,and invasion of HepG2 cells by inhibiting the target mRNA.4.Sorafenib affects the expression of LncRNA,which affects the biological behavior of HepG2 cells by regulating mRNA.5.Sorafenib regulates CircRNA,and CircRNA acts as a miRNA sponge,which binds a large number of miRNAs competitively with mRNA,thereby inhibiting the negative control of miRNAs on their target mRNAs and thus affecting the function of HepG2.