Expression Of SPC24 In Prostate Cancer And Its Significance In Combination With BUB1 And NDC80 In Diagnosis Of Prostate Cancer

Objective: SPC24 is a subunit of the Nuclear Division Cycle 80(NDC80)complex,which is involved in the formation of the NDC80 complex with three other proteins(NUF2,NDC80,and SPC25).With the help of the NDC80-NUF2 protein complex,the NDC80 complex mediates microtubule binding.Further,the NDC80 complex is anchored onto the inner kinetochore via the SPC24-SPC25 protein complex.In addition,SPC24-SPC25 protein complex mediates dynamic interactions between the nuclear spindle microtubules and kinetochores,which ensures faithful and accurate chromosomal segregation during mitosis At present,a large number of studies have found that SPC24 is highly expressed in various cancers,but its effect in prostatitis,Prostatic Carcinoma(PCa)and Benign Prostatic Hyperplasia(BPH)has not been reported.This research group intends to study the expression of SPC24 gene in prostatitis,PCa,and BPH and explore the diagnostic value of SPC24 and its interaction model to PCa.We will search for new biomarkers for the diagnosis and prognosis of PCa in the future.Methods: 1.RNA-Seq data from The Cancer Genome Atlas(TCGA)database were used to analyze SPC24 messenger RNA(m RNA)levels in PCa tissues and normal tissues.And we discussed the SPC24 m RNA expression,clinicopathologic characteristics analysis and survival analysis.2.Collect clinical tissue samples,extract tissue RNA and tissue protein respectively;use real-time quantitative polymerase chain reaction(RT-q PCR)and Western blot(WB)to detect the expression of SPC24 in PCa tissue,adjacent tissue and BPH tissue.The expression of SPC24 in PCa and adjacent tissues,BPH tissue,prostatitis and normal tissues was detected by immunohistochemistry(IHC),and the association between SPC24 expression and clinicopathological characteristics of PCa /BPH / prostatitis patients were assessed.3.SPC24 interactors were retrieved by string online analysis software(http://string-db.org/),and SPC24 and its interactors(NDC80,BUB1)were analyzed by Gene ontology(GO)analyses and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway.RT-q PCR was used to verify the m RNA expression levels of BUB1 and NDC80 in PCa and adjacent tumors.Based on m RNA data(quantified using RSEM)in the TCGA database,two combined models(SPC24 + BUB1,SPC24 + NDC80)were established.Using binary logistic regression,the ROC curve was used to analyze the diagnostic value of two combined models for PCa.Results: 1.m RNA expression data of 550 prostate samples downloaded from TCGA database,including 498 PCa samples and 52 normal prostate samples were included in the study.This data were analyzed by independent sample t test,and the results showed that compared with normal prostate tissue,the m RNA expression of SPC24 in PCa tissue was significantly higher than that in normal prostate tissue(P <0.0001).And a number of indicators are of clinical significance.Among them,PCa patients with higher Gleason scores(8-10)and higher PSA value(> 0.1)were found to have high expression of SPC24.Patients with higher SPC24 expression have shorter OS in survival analysis.And the AUC value was 0.821(p<0.05).2.IHC results showed that in the PCa / adjacent group,37.14%(13/35)of PCa patients had high expression of SPC24,and the rest showed low / negative expression(P<0.0001);in the BPH / adjacent group,27.5%(11/ 40)of BPH patients had high expression of SPC24,and the rest showed low / negative expression(P=0.001);in the prostatitis / normal group,41.67%(5/ 12)of patients with prostatitis showed high expression of SPC24,and the rest showed low / negative expression(P=0.045);and SPC24 was in the adjacent tissues(35/35)and normal tissues(9/9)both showed low / negative expression(P<0.05);There was no difference between the expression of SPC24 in the PCa / BPH group(P>0.05).The high expression of SPC24 in patients with prostate cancer was related to Gleason stage(Ⅳ+Ⅴ)(P <0.05);The expression of SPC24 in patients with BPH was related to nationality and Cr Cl(P <0.05).There was no association between SPC24 expression and other clinical factors in patients with prostatitis.WB results showed that the protein expression of SPC24 in PCa tissues was significantly higher than that in adjacent tissues(P<0.05).RT-q PCR analysis showed that SPC24 m RNA level was significantly higher in PCa than AT(8.43 ± 3.00 vs 1.22 ± 0.25,P <0.05);it was significantly higher in PCa than BPH(8.43 ± 3.00 vs 4.09 ± 1.72,P <0.05);in BPH it was higher than AT Significantly increased(4.09 ± 1.72 vs 1.22 ± 0.25,P <0.05).That is,the expression level of SPC24 m RNA was highest in PCa,second in BPH,and lowest in AT,and the difference was statistically significant.3.The STRING database performed GO gene annotation and KEGG pathway enrichment analysis on SPC24 and its interacting proteins(BUB1,NDC80).The GO analysis results showed that SPC24 and its interacting proteins are mainly involved in the cell cycle,cell division process,and important groups of cells.In KEGG pathway analysis,SPC24 and its interacting proteins are involved in the separation of sister chromatids,MAD2 pathway,cell cycle checkpoints,small GTPase activity Activate formins,and signal transduction.RT-q PCR results showed that the expressions of BUB1 and NDC80 were higher in PCa tissues than in adjacent tissues.The results indicate that the m RNA expression of SPC24 and its interactors is positively correlated with the prevalence of PCa.The ROC curve of combined model of SPC24 and BUB1 was generated,and the sensitivity and specificity were 88.2% and 80.8%,respectively.The AUC was0.894.The ROC curve of combined model of SPC24 and NDC80 had a AUC of0.905,and the sensitivity and specificity were 84.7% and 96.2%,respectively.The AUC of the two combined models(SPC24 + BUB1,SPC24 +NDC80)in the ROC curve were>0.89(P <0.05).Conclusions: 1.TCGA database and tissue verification show that SPC24 is highly expressed in prostate cancer tissues.2.The high expression of SPC24 is associated with the low survival of prostate cancer patients.3.The high expression of SPC24 in prostate cancer is related to Gleason score and t PSA;the high expression in prostate hyperplasia samples is related to race and Cr Cl.4.Combined diagnosis models have higher diagnostic significance in prostate cancer.