| Prostate cancer is a common malignant tumor of urinary system,with a rapid increase in morbidity and mortality.Its occurrence and development are affected by a variety of complex factors,but its pathogenesis has not been clearly defined.Androgen deprivation therapy(ADT)is the main means of treating advanced prostate cancer,which plays a role in alleviating the progression of the tumor.However,drug resistance eventually develops into castration-resistant prostate cancer,which is the main cause of poor prognosis and death of patients.In recent years,gene targeting therapy has played an obvious role in tumor therapy.More and more studies have shown that KIF2 C plays an important role in the occurrence and development of various tumors,but it has not been reported in prostate cancer.This study aims to find out the expression characteristics,role and mechanism of KIF2 C in prostate cancer through bioinformatics analysis,and further explore its influence on survival and prognosis,as well as study the influence of KIF2 C on biological function of prostate cancer cells through cell experiments.Objective(s):To comprehensively study the characteristics of KIF2 C in prostate cancer and its effects on the proliferation and migration of prostate cancer cellsMethods:1.Bioinformatics analysis: Combined with large-scale transcriptome data,including databases such as the Cancer Genome Atlas,cancer cell line Encyclopedia,genotypic tissue expression,c Bio Portal,and cancer drug sensitivity genomics,The association of KIF2 C with prognosis,clinicopathological features,gene mutation,DNA methylation,tumor microenvironment,immune checkpoint,immune cell infiltration,and drug resistance of prostate cancer was analyzed by a series of bioinformatics methods.2.Validation of clinical specimens: Cancer tissues and corresponding paracancer tissue specimens of prostate cancer patients were collected for RT-qPCR analysis to verify the expression level of KIF2 C in prostate cancer tissue specimens.3.Validation of clinicopathologic sections: The pathological sections of cancer tissues and corresponding paracancer tissues of prostate cancer patients were collected for IHC analysis to evaluate the expression level of KIF2 C in prostate cancer tissues.4.Cell experiment verification: human prostate cancer cells PC3,LNCAP and human normal prostate epithelial cells RWPE-1 were cultured,and RT-qPCR and WB experiments were used to evaluate the difference in the expression of KIF2 C in prostate cancer cells and normal prostate cells.5.The transfection cells were divided into the following groups:(1)Transfection group: PC3 and LNCAP cell lines transfected with KIF2 C knockdown low plasmid.(2)Idle group: PC3 and LNCAP cell lines transfected with KIF2 C no-load plasmid.6.Effect of KIF2 C on biological function of prostate cancer cells: We established the above transfection group cell lines and idle group cell lines for routine culture.The OD450 values of the two groups of cell lines at 0h,24 h,48h and 72 h were measured by CCK8 method to detect the proliferation.Ed U method was used to measure the fluorescence quantity and positive rate of the two groups of cell lines to detect the difference in proliferation.The Transwell method was used to compare the number of cell lines in the two groups that migrated through Transwell compartments to the subpore-membrane cells to detect migration.Results:1.The expression level of KIF2 C in human tissues was evaluated with GTEx database,and it was concluded that KIF2 C was highly expressed in bone marrow and testis tissues,while generally low in other tissues.KIF2 C was highly expressed in various tumor tissues using CCLE database.The TCGA database was further used to analyze the pan-cancer data.Among the 33 tumors,the expression level of KIF2 C in28 tumors was higher than that in the corresponding adjacent tissues.2.The results of clinicopathological analysis of KIF2 C and prostate cancer indicated that the expression level of KIF2 C in 52 pairs of tumor samples was higher than that in corresponding adjacent tissues,and the expression level of KIF2 C was also increasing with the continuous increase of T stage,lymph node metastasis,distant metastasis,PSA and Gleason score.Meanwhile,KIF2 C levels were relatively high in prostate cancer patients aged 60 years or older.Analysis of TCGA-PRAD cohort data showed that the expression level of KIF2 C was correlated with T stage,lymph node metastasis,PSA and Gleason score.3.The results of prognostic analysis of KIF2 C and prostate cancer indicated that the expression of KIF2 C was significantly correlated with overall survival(OS),disease-free interval(DFI),disease-specific survival(DSS)and progression-free interval(PFI),and was a high-risk gene.Kaplan-Meier analysis showed that increased KIF2 C expression level was highly correlated with poor OS,DFI,DSS and PFI.Meanwhile,ROC curve analysis results showed that KIF2 C could be used as an effective prognostic marker for prostate cancer,and the accuracy of prognostic analysis was higher than PSA.4.The expression level of KIF2 C was negatively correlated with promoter methylation in many mutant genes.With different expression levels of KIF2 C in prostate cancer,the types of mutated genes were significantly different.The results of functional enrichment analysis of KIF2 C in prostate cancer indicate that KIF2 C is involved in cell cycle,and plays an important role in megakaryocyte development,platelet generation,cell stress,HIV infection,and adaptation to the immune system.5.Correlation analysis between KIF2 C expression level and tumor microenvironment showed that high expression level of KIF2 C was significantly correlated with immune-related features and mismatch repair related features.At the same time,KIF2 C expression level was positively correlated with immune score and estimated score,and negatively correlated with tumor purity.6.KIF2 C has positive or negative correlation with immunoinfiltration of many tumor cells.In prostate cancer,KIF2 C is positively associated with almost all immune-related genes and is strongly associated with multiple other immune checkpoint members.Meanwhile,high expression of KIF2 C is involved in drug resistance of various MAPK signaling pathway inhibitors.7.In prostate cancer tissues,the results of RT-qPCR and immunohistochemical detection showed that the expression level of KIF2 C was significantly higher than that of paracancer tissues(P<0.01).In prostate cancer cells,RT-qPCR and WB detection showed that KIF2 C expression level was higher than that of human normal prostate epithelial cell lines(P<0.01).8.The results of CCK8 and Ed U suggested that compared with the idle group,the cells transfected with KIF2 C knockdown low plasmid significantly inhibited the proliferation of PC3 and LNCAP prostate cancer cells(P < 0.01).The Transwell results suggested that the cells transfected with KIF2 C knockdown low plasmid inhibited the migration of PC3 and LNCAP prostate cancer cells(P<0.001).Conclusion(s):In this study,it was confirmed that KIF2 C can be specifically expressed in prostate cancer,and the expression level of KIF2 C in prostate cancer tissues is higher than that in adjacent tissues.KIF2 C expression level in prostate cancer was positively correlated with age,stage,grade,PSA,Gleason score,lymph node metastasis and distant metastasis.The higher KIF2 C expression level,the worse the prognosis.KIF2 C expression level was negatively correlated with promoter DNA methylation,positively correlated with immune score and estimated score,and closely correlated with a variety of immune cell infiltration,and correlated with tumor microenvironment,immune infiltration and immune checkpoint genes.In addition,the high expression of KIF2 C led to increased drug resistance of multiple MAPK signaling pathway inhibitors.The level of KIF2 C expression in prostate cancer cells is higher than that in normal prostate epithelial cells,and the proliferation and migration of prostate cancer cells can be inhibited by targeting KIF2 C knockdown. |
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