Study On The Mechanism And Roles Of Melk Signaling Pathway On Hep-G2 And Hep-3B Cells In Hepatocellular Carcinoma

Background and aims Hepatocellular carcinoma(HCC)is the sixth most common cancer in the world.In recent years,medical concepts,diagnosis and treatment techniques have been continuously innovated,and surgical treatments,local ablation,hepatic vascular embolization,radiotherapy,local chemotherapy,and targeted therapy have all made progress,showing a comprehensive treatment plan based on surgery.So far,although MELK as the main target inhibitor has not been approved by the FDA,various MELK inhibitors have been actively researched in recent years.Perhaps because the function and mechanism of MELK in tumor formation have not been fully revealed.And A variety of genetic variation factors and environmental factors may involve in the development of tumors,so that MELK inhibitors have not been effectively applied to various clinical trials.Therefore,a comprehensive understanding of the functional characteristics of MELK on malignant tumor-associated carcinogenic kinases can promote the research of small molecule kinase inhibitors targeted therapy for malignant tumors,which will be the research direction in the next few years.Materials and methods1.Oncomine and Kaplan-Meier plotter database for deep excavation of hepatoma MELK gene information2.Expression of MELK in different tumors3.mRNA expression and detection of MELK co-expressing genes in different tumors4.Verification of MELK interference efficiency5.Effect of MELK siRNA interference on cell proliferation6.Effect of OTPSSP167 inhibitor on proliferation,inhibition,viability and IC50 value of hepatoma cells7.Effect of OTPSSP167 inhibitor and SiRNA interference on tumor cell invasion ability8.Effect of OTSSP167 inhibitor and SiRNA interference on tumor cell clonal proliferation9.Effect of OTSSP167 inhibitor and SiRNA interference on apoptosis of hepatocellular carcinoma Hep-3B Results1.MELK gene is highly expressed in various tumors,including hepatocellular carcinoma,breast cancer,colorectal cancer,etc.gene expression is related to the survival state of patients.genes co-expressed by MELK include: TOP2 A,RACGAP1,GINS1,NDC80,etc.2.The expression of Hep-3BMELK mRNA in hepatoma cells is higher than that in PANC-1 cell line,HGC27 and Hep-G2 cell lines.3.HEP-3B,MCG-27,PANC-1 cell line MELK co-expression genes are GINS1,NDC80,RACGAP1.4.The expression of MELK SiRNA1,MELK SiRNA2,MELK SiRNA3,and MELK mRNA decreased after SiRNA interference.5.There was no significant difference in proliferation ability between SiRNA1 group,SiRNA2 group,SiRNA3 group,GAPDH group and Mock Transfection group compared with the control group.6.With the increase of the concentration of OTSSP167 inhibitors,the proliferation and viability of cell lines decreased gradually,and the inhibition rate of cell lines gradually increased.The IC50 values of Hep-3B and Hep-G2 cell lines were 222 nM and 101.3nM,respectively,and the 95% confidence intervals were:(198-249.8)nM,(74.42-141.1)nM7.OTSSP167 inhibitors had significant effects on the invasive ability of Hep-3B and Hep-G2,and the cell invasion ability at 48 hours and 72 hours decreased compared with the control group(P<0.001).SiRNA interference had no effect on the invasive ability of hepatocellular carcinoma Hep-3B and Hep-G2,and the cell invasive ability at 48 hours and 72 hours did not change significantly compared with the control group(P>0.05).8.After the expression of MELK was disturbed,the proliferation ability of hepatoma cells in vitro was not significantly decreased(P>0.05),while the OTSSP167 inhibitor group significantly inhibited the in vitro colonization of hepatoma cells(P<0.001).9.Compared with the control group,no early apoptotic cells were observed in the SiRNA interference group;the number of early apoptotic cells increased significantly in the OTSSP167 group with the increase of inhibitor concentration.Conclusions(1)The high expression of MELK gene in Hep-G2 and Hep-3B cells is related tobiological functions such as apoptosis,proliferation and migration of tumor cells.(2)SiRNA interferes with MELK gene expression,and the biological functions ofHep-G2 and Hep-3B cells are not inhibited.(3)OTSSP167 inhibitors can inhibit the proliferation and migration of Hep-3B andHep-G2 clones,and reduce the anti-apoptotic ability of tumor cells.With the increase ofinhibitor concentration,OTSSP167 inhibitors can inhibit the biological function of tumor cells.