TTK/Mps1 Promotes Prostate Cancer Growth And Progression

Objective Bioinformatics is a discipline that studies the collection,processing,storage,transmission,analysis and interpretation of biological information.The research focus is mainly reflected in Genomics and Proteomics,that is,starting from nucleic acid and protein sequences,the analysis of the biological information of structural functions expressed in the sequences.We detected that the mRNA level of TTK gene through TCGA database increased with the increase of clinical Gleason score,so we speculated that TTK played an important role in the development of prostate cancer.By searching relevant literatures,it was found that TTK had little research on the progress of prostate cancer.Therefore,we verified the expression of TTK in prostate cancer cell lines through a series of experiments and explored the changes in its biological behavior.Methods According to the pathological grading of prostate cancer,immunohistochemical staining was performed on pathological sections of prostate cancer with different Gleason scores to verify TTK expression in prostate hyperplasia and prostate cancer tissues with Gleason scores of 6 and 9.MRNA and total protein of the four cell lines were extracted by cultured LNCaP,C42,DU145 and PC3 prostate cancer cell lines in vitro,and the mRNA level and protein expression of TTK in the four cell lines were verified by q-PCR and Western Blot.In order to investigate whether the deletion of TTK gene affects the proliferation,invasion and metastasis of prostate cancer,we designed specific siRNA sequences of TTK gene to knock out the expression of TTK.The migration,invasion,proliferation and apoptosis levels of PC3 cell lines after TTK gene knockout were verified by cell scratch test,Transwell test,CCK8 test and cell flow test,respectively.The lentivirus transfected with shRNA sequence was cultured with PC3 cell lines to form stable cell lines for passage,and the in vivo experiment was conducted to verify whether the knockout of TTK gene had an effect on the tumorgenesis of prostate cancer.Result The results showed that the expression of TTK gene in prostate cancer hyperplasia and prostate cancer tissues with different Gleason scores gradually increased.The expression of TTK was verified by mRNA and protein levels of 4 cell lines in vitro.It was found that the expression of TTK in metastatic cell lines such as DU145 and PC3 cell lines was significantly higher than that in non-metastatic cell lines such as LNCaP and C42.The experimental results of biological behavior further demonstrated that after TTK knockout,PC3 cells’ migration,invasion,proliferation and apoptosis degree increased,and G1 phase arrest significantly increased while G2 phase decreased.The results of in vivo experiments also showed that the rate of subcutaneous tumorigenesis of nude mice significantly decreased after TTK knockout,and there was no tumorigenesis in the co-culture time.Conclusion 1.Immunohistochemistry,q-PCR and Western Blot jointly verified that TTK expression level was related to the progression of prostate cancer.2.The migration and invasion abilities of PC3 cells were positively correlated with TTK expression 3.The results of cell flow experiment showed that after TTK knockout,G1 and S phase blocks increased,G2 phase decreased,and apoptosis level increased 4.In vivo test results showed that the tumorigenesis rate of PC3 cells transfected with TTK knockout was significantly lower than that of the control group.