A Preliminary Study On The Role And Mechanism Of CDKL3 In Mediating The Occurrence And Development Of Hepatocellular Carcinoma

Research purpose Tumor is a kind of cell periodic disease.If the cell initiation process and signal transduction pathway fail,the cell cycle process may be out of control,resulting in abnormal cell proliferation,and eventually may develop into tumor.Sample cycle dependent kinase protein 3(CDKL3)protein kinase dependent sample is cycle(CDKL)a member of the family.In recent years,some studies have found that CDKL3 level adjustment with osteosarcoma,esophageal squamous cell carcinoma is closely related to the cell cycle and cell proliferation,however CDKL3 in hepatocellular carcinoma(HCC)in biological and clinical significance is not reported.Hepatocellular carcinoma(HCC)is the most common histological type of liver cancer and one of the most common cancers,ranking fourth among cancer-related deaths in men in China.Given CDKL3 structure,dependence and cell cycle protein Kinase(Cyclin-dependent Kinase,CDK)and Mitogen activated protein Kinase(Mitogen-activated protein Kinase,MAPKS)of homologous sequences,and CDKL3 and some closely related to the occurrence and development of other tumors have been studies confirm,so we make assumptions CDKL3 may adjust the hepatocellular carcinoma cell cycle and drive the occurrence and development of hepatocellular carcinoma(HCC),provide new way for treatment of liver disease.Methods 1.CDKL3 m RNA expression in 3 human liver cancer cell lines and normal human liver cells was detected.2.Lentivirus infected hepatoma cell line Hep G2 and SMMC-7721,Huh-7 silenced the expression of CDKL3 in hepatoma cell line Hep G2.The infection efficiency was observed under inverted fluorescence microscope,and the knockdown efficiency of CDKL3 was detected by q RT-PCR and Western blotting.3.CCK8 cell proliferation assay,clone formation assay to detect the proliferation ability of liver cancer cell lines after CDKL3 silencing;Transwell migration assay and cell scratch assay were used to detect the migration ability of HCC cell lines after CDKL3 silencing.4.The apoptosis rate of HCC cell lines after CDKL3 silencing was detected by TUNEL one-step assay;Cell cycle assay was used to detect the cell cycle of HCC cell lines after CDKL3 silencing.Subcutaneous tumorigenesis assay in nude mice was used to detect the ability of CDKL3 to silence hepatocellular carcinoma in vivo.5.The protein expression of ERK1/2,p-ERK1/2,Akt and p-Akt after CDKL3 silencing was detected by Western blotting.Results 1.The m RNA content of CDKL3 in the three hepatoma cell lines was higher than that of normal human hepatoma cell lines.2.The infection efficiency of lentivirus in liver cancer cell lines was more than 80%.After infection,CDKL3 m RNA content and protein expression in liver cancer cell lines were decreased.3.The results of CCK8 and clone formation experiments showed that the proliferation ability of HCC cell lines was significantly reduced after CDKL3 knockdown.Transwell and scratch experiments showed that CDKL3 knockdown significantly reduced the migration ability of HCC cell lines.4.After CDKL3 knockdown,the apoptosis rate of HCC cell lines reached 90% and the cell cycle was arrested.Tumor-forming experiments in nude mice showed that the tumor-forming ability of CDKL3 knockdown hepatocellular carcinoma cells in nude mice was significantly reduced,the tumor volume was smaller,and the growth rate was slower.5.Under the condition of the same expression of ERK1/2 protein,the expression of p-ERK1/2 protein decreased after CDKL3 silencing.conclusion 1.CDKL3 can mediate the proliferation,migration,apoptosis,cycle and other biological behaviors of HCC cells.2.CDKL3 may mediate the occurrence and development of hepatocellular carcinoma through ERK pathway.