The Role Of CDK11^p110 In The Development Of Esophageal Squamous Cell Carcinoma

Introduction and aimsEsophageal squamous cell carcinoma（ESCC）is one of the main pathological modality of esophageal carcinoma,with high incidence,high mortality and poor prognosis,there is a lack of effective treatment in clinic.Cyclin-dependent kinase（CDK）is an important regulatory factor in cell cycle,and drug development targeting CDK has become a promising approach for tumor treatment.Previous studies have shown that CDK11^p110 plays an important role in the occurrence and development of various tumors,but its role in esophageal cancer has not been reported.This paper aims to study the mechanism of CDK11^p110 in the occurrence and development of ESCC,and to explore the potential of targeted CDK11^p110 in the treatment of ESCC,in order to provide a new direction for the treatment of ESCC.MethodsIn this study,firstly,the expression of CDK11 in esophageal carcinoma was analyzed using bioinformatics in the TCGA data set,the expression of CDK11^p110and the localization of CDK11^p110 in ESCC were detected by immunohistochemistry,and the expression of CDK11^p110 in clinical ESCC tissues and cells was detected by Western blot,and the expression of CDK11^p110 in ESCC was preliminarily analyzed;Secondly,interference with the expression of CDK11^p110 in ESCC cells by siRNA,the interference efficiency of CDK11^p110 was detected by immunofluorescence and Western blot,and effects of CDK11^p110 on proliferation,clonal formation and migration of ESCC cells were detected by CCK8,clonal formation and Transwell methods,to explore the role of CDK11^p110 in the development of ESCC;Thirdly,after interfering with CDK11^p110,Western blot was used to detect the changes in apoptosis adhesion growth immunity and other related signaling pathway proteins in ESCC cells,and flow cytometry was used to detect the changes in apoptosis and cell cycle arrest of ESCC cells,so as to further elucidate the mechanism of CDK11^p110promoting the occurrence and development of ESCC;Finally,construction of a stable ESCC cell line that overexpresses and knockdown CDK11^p110 by lentiviral vector,CCK8 was used to detect the effect of stable knockdown and overexpression of CDK11^p110 on the growth and proliferation of ESCC cells,so as to investigate the regulatory mechanism of CDK11^p110 in ESCC at subsequent in vivo levels.ResultsmRNA expressions of CDK11A and CDK11B were significantly higher in esophageal cancer tissues than in adjacent tissues,and immunohistochemical analysis showed that CDK11^p110 expression was significantly higher in ESCC tissues than in adjacent tissues,and CDK11^p110 is mainly located in the nuclei of ESCC cells,the expression of CDK11^p110 protein in human ESCC was significantly higher than that in adjacent tissues,and the expression of CDK11^p110 protein in 6 human ESCC（EC109,EC9706,KYSE150,KYSE70,TE-1 and TE-7）was significantly higher than that in normal esophageal epithelial cells（HET-1A）.The expression of CDK11^p110 in EC109 and KYSE150 cells transfected with siRNA was significantly decreased,moreover,siRNA CDK11^p110 not only significantly inhibited the growth of EC109and KYSE150 cells in a dose-dependent and time-dependent manner,but also inhibited the ability of cell clone formation and migration.Interfering with CDK11^p110 can upregulate the expression of apoptosis-related protein PARP,arrested EC109 and KYSE150 cells in the G2/M phase,and induced apoptosis of cells.The stably transfected EC109 and KYSE150 cell lines with overexpression and knockdown of CDK11^p110 were successfully constructed,the expression of CDK11^p110 in stably transfected EC109 and KYSE150 cell lines was significantly increased,while the protein level of CDK11^p110 in the two stably transfected low cell lines was significantly decreased,consistent with the previous experiments,the activity of EC109 and KYSE150 cell lines with stable knockdown of CDK11^p110 was significantly inhibited,while the proliferation of EC109 and KYSE150 cells with overexpression of CDK11^p110 was increased.ConclusionsCDK11^p110 is highly expressed in human ESCC tissues and cell lines,and CDK11^p110 is mainly located in the nuclei of ESCC cells.Interfering with the expression of CDK11^p110 can significantly inhibit the proliferation,colony formation and migration of ESCC cells,increase the expression of apoptosis-related protein PARP,and induce cell G2/M phase arrest and apoptosis.Successfully constructed stable ESCC cell lines that overexpresse and knock down CDK11^p110,and the overexpression of CDK11^p110 promoted the proliferation of ESCC cells.