Effect Of KLF4 On Ovarian Cancer Cell Behavior

BackgroundOvarian cancer is a common female reproductive system malignant tumor.For the most part,it presents with non-specific symptoms and is usually diagnosed at an advanced stage due to limited screening methods and medical treatment.Therefore,the highest mortality rate in ovarian cancer in female reproductive system disease.The main treatment options for ovarian cancer include surgery and chemotherapy,although significant advances in current treatments,the majority of patients with advanced ovarian cancer still experience recurrence and metastasis,and eventually become resistant to chemotherapy drugs.Thus,exploring the emergence of ovarian cancer,the molecular mechanism of progress and metastasis has great significance on the diagnosis and treatment of ovarian cancer.KLF4,a member of the "Krüppel" family,has a relationship with a variety of cancer.KLF4 has transcriptional activation and inhibition domains,which can positively and negatively regulate downstream targeted genes and participate in the occurrence and development of tumors.There are few studies on the expression and biological function of KLF4 in ovarian cancer.At present,there are few studies on the expression of KLF4 in ovarian cancer and its biological function,so this study uses the database to analyze the effects of KLF4 in the emergence and development of ovarian cancer,and further investigate the functional effect of KLF4 on ovarian cancer cells by cytology experiments,and initially explores the molecular mechanism of KLF4 on ovarian cancer.ObjectiveIn this study,the expression of KLF4 in ovarian cancer was analyzed by bioinformatics database,and the expression of KLF4 gene in ovarian cancer cell lines and the behavioral effects on ovarian cancer cell lines were detected by in vitro cell experiments.Provide a basis for the early diagnosis and management of ovarian cancer in the clinic.Method1.GEPIA database was used to analyze the expression of KLF4 in normal ovarian tissues and ovarian cancer tissues.For follow-up experiments,the mRNA expression of KLF4 in different human ovarian cancer cell lines were analyzed by CCLE,and the mRNA expression of KLF4 were verified by PCR.2.In order to study the biological impact of KLF4 on ovarian cancer cell lines,A2780 cell line with low KLF4 expression and HeyA8 cell line with high KLF4 expression were selected for in vitro function study.The overexpressed plasmid was transfected into A2780 cell line to obtain stable overexpressed KLF4 cell line.The shRNA was packaged with lentivirus and infected with HeyA8 cell line to obtain KLF4 knockout stable cell line.The mRNA and protein expression of KLF4 in A2780 and HeyA8 ovarian cancer cell lines were verified by Western Blot and qPCR.3.The effect of KLF4 on the in vitro proliferation ability of ovarian cancer cell lines was analyzed by CCK-8;the effect of KLF4 on the migration ability of ovarian cancer cell lines was analyzed by cell migration experiments;the effect of KLF4 on the invasion ability of ovarian cancer cell lines was analyzed by cell invasion experiments.4.The UCSC public database was used to conduct GSEA,and analyze the pathways involved in the down-regulation and up-regulation of KLF4.5.Downloading the sequencing results of KLF4 knockdown in human keratinocytes from the GEO database,looking for genes that are differentially expressed.PCR analyzes the expression of these genes in ovarian cancer cells.Result1.GEPIA analysis showed that the expression level of KLF4 gene in some common tumors is different,compared with normal ovarian tissue,KLF4 expression in cancerous tissue is low.2.Through the analysis of CCLE,the expression of KLF4 mRNA in different human ovarian cancer cells was different,and the KLF4 mRNA expression of various ovarian cancer cells was verified by PCR,and the results were consistent with the CCLE.3.The qPCR and Western Blot were used to verify that overexpression of KLF4 in A2780 cell line increased the mRNA and protein expression of KLF4.After knockdown the expression level of KLF4 in HeyA8 cell line,the mRNA and protein expression of KLF4 were decreased.4.Cell proliferation experiments show that KLF4 has the effect of inhibiting cell growth in ovarian cancer cell lines;migration experiment results show that KLF4 has the ability to inhibit the migration of ovarian cancer cells;invasion experiment results can see that KLF4 has the ability to inhibit the invasion of ovarian cancer cells.5.GSEA showed that gene sets of ETS pathway,INTEGIN pathway,p27 pathway,ATRBRCA pathway,ETC pathway and SET pathway were enriched in low-expression group and high-expression group of KLF4.6.14 differentially expressed downstream genes were obtained by GEO analysis.The PCR showed that overexpressing KLF4 in the A2780 cell line,the expression of RAB39 B and TBX2 was down-regulated and MYEOV was up-regulated,which was consistent with the results of GEO data analysis.ConclusionCompared with normal ovarian cancer tissues,the KLF4 is lowly expressed in ovarian cancer tissues and ovarian cancer cell lines.And KLF4 may affect cellular behavior in ovarian cancer by participating in cell cycle regulation related pathways and regulating the expression of downstream genes RAB39 B,TBX2 and MYEOV.