KLF13 Promotes Proliferation、Invasion And Metastasis Of Hepatocellular Carcinoma Cells By Up Transcriptional Regulation HMGCS1

Objective:Krüppel-like factor(KLF)plays an important role in the occurrence,development and metabolism of cancer.We aimed to explore the role and potential molecular mechanism of KLF13,a member of Krüppel like factor family,in the growth and migration of hepatoma cells.Methods:The m RNA expression data set was downloaded from the Cancer Genome Atlas Database(TCGA),and the expression of KLF13 in hepatocellular carcinoma was analyzed by R language.RT-q PCR and Western blot were used to detect the m RNA and protein expression levels in liver cancer tissues,adjacent tissues and cell lines.Overexpression of KLF13 in SK-Hep1 and Huh7 hepatoma cell lines with lentivirus vector and knockdown of KLF13(sh Ctrl and sh KLF13)in Hep G2 and MHCC-97 H cell lines with plasmid.Cell proliferation was analyzed by cell counting kit-8(CCK-8),plate cloning,Ed U,and flow cytometry.The effects of KLF13 on the migration and invasion of hepatoma cells were analyzed.Subcutaneous xenograft tumor in nude mice to evaluate the effect of KLF13 on the growth of nude mice.The content of free cholesterol in liver cancer cells overexpressing KLF13 or vector and treated with sihmgcs1 or sicrl was measured by cholesterol indicator kit.Ch IP-seq revealed that the target gene downstream of KLF13 was hmgcs1.The double luciferase reporter gene revealed the binding relationship between KLF13 and HMGCS1 promoter.Results:KLF13 was found to be highly expressed in hepatocellular carcinoma tissues in TCGA database.RT-q PCR and Western blot analysis showed that KLF13 m RNA and protein levels were up-regulated in hepatocellular carcinoma compared with adjacent tissues.Functional experiments showed that KLF13 gene knockdown inhibited the proliferation,migration and invasion of Hep G2 and Huh7 cells.Overexpression of KLF13 promotes the proliferation,migration and invasion of SK-Hep1 and MHCC-97 H cells.In addition,KLF13 gene knockdown increased the apoptosis rate of Huh7 cells and decreased the expression of cyclin D1,CDK4 and CDK2.KLF13 knockdown reduced the transformation of Huh7 cell cycle from G1-G2 / M phase and increased the expression of anti apoptotic protein BCL2.KLF13 overexpression decreased the apoptosis rate of SK-Hep1 cells and increased the transformation of cell cycle from G1-S phase.Overexpression of KLF13 promoted the growth of subcutaneous liver cancer in nude mice.Further experiments showed that KLF13 transcription promoted the expression of HMGCS1 and cholesterol biosynthesis.Ch IP-seq and double luciferase reporter genes revealed that the transcription factor KLF13 directly binds to the promoter of the downstream target gene HMGCS1.KLF13 gene knockdown inhibits HMGCS1 mediated cholesterol biosynthesis and inhibits the growth and metastasis of hepatoma cells.On the contrary,si HMGCS1 was added to SK-Hep1 and MHCC-97 H cells overexpressing KLF13 at the same time.Functional experiments showed that si HMGCS1 reversed its promotion of proliferation and migration of hepatoma cells.Conclusions:1.Overexpression of KLF13 promotes the proliferation and migration of hepatoma cells,while low expression of KLF13 inhibits the proliferation,migration and other malignant biological behaviors of hepatoma cells.2.KLF13 promotes proliferation 、 invasion and metastasis of hepatocellular carcinoma cells by up transcriptional regulation HMGCS1...