The Role Of KNTC1 On The Regulation Of The Development,Invasion And Metastasis In Cervical Carcinoma

BackgroundCervical cancer is a common malignant tumor that threatens women’s health.It ranks second in the most commonly diagnosed tumors in the female population.The incidence of cervical cancer in China accounts for about 1/3 of the total number of people in the world.It has become an important cause of cancer death in women and poses a serious threat to the health of women in China.The occurrence and development of cervical cancer is closely related to the infection of human papillomavirus(HPVs),especially,HPV 16 and HPV 18 are the two main subtypes that cause cervical cancer after most HPV infections.The early proteins they encode,E6 and E7,are key proteins that lead to malignant transformation of cervical cells.E6 can inhibit P53 and E7 degrade pRb,respectively,causing the infected cells to unlimited growth and malignant transformation.However,only a small number of cases have progressed to malignant lesions after high-risk HPV infection.HPVs infection is only a sufficient but not a necessary condition for the occurrence of cervical cancer.The invasion and metastasis of cervical cancer may not depend on E6 or E7.The mechanism of occurrence and development of cervical cancer remains to be further elucidated.Kinetochore associated 1(KNTC1),or the rough deal(ROD)gene,which is located on human chromosome 12,and its encoded protein is an important component of mitotic checkpoints.This protein participates in a variety of mechanisms to ensure the proper separation of chromosomes during mitosis.KNTC1 is highly expressed in cervical cancer,suggesting that KNTC1 may be involved in the development of cervical cancer,but its role in cancer is still unclear and needs further study.ObjectivesThis paper intends to preliminarily clarify the expression of KNTC1 in cervical cancer,the influence of the invasion and metastasis of cervical cancer and its possible regulatory mechanism.Methods1.The relative expression of KNTC1 was determined by real-time fluorescence quantitative PCR(RT-qPCR)after the total RNA was extracted and reversed to cDNA.The differences in the expression of KNTC1 gene between cervical and cervical cancer tissues and different cell lines were compared.2.Cervical cancer cell lines HeLa and SiHa were transiently transfected with siRNA to knock down the expression of KNTC1.The effect of KNTC1 on cell migration and invasion was detected by Transwell migration and invasion assay;the effect of KNTC1 on cell proliferation was detected by EdU kit and plate colony formation assay.3.After silencing the expression of KNTC1,the Western Blot method was used to detect the change of the biological markers of tumor epithelial-mesenchymal transition(EMT)related molecules at the protein level.4.The expression of KNTC1 in HeLa cells was silenced,mRNA expression microarray was used to detect differential gene expression,and KEGG pathway enrichment analysis was used to select downstream target genes that may be directly or indirectly controlled by KNTC1,in order to search for possible regulatory mechanisms.Results1.The results of qPCR showed that compared with the normal cervical tissue,the expression of KNTC1 in cervical cancer tissues was higher and KNTC1 was expressed in 3 cervical cancer cell lines.2.After silencing the expression of KNTC1,the results of Transwell showed that the migration and invasion ability of KNTC1 knockdown group were significantly increased.The results of EdU showed that the proliferation ability of KNTC1 interference group was significantly enhanced and the ability of clone formation increased significantly.3.After siRNA inhibited the expression of KNTC1,the expression of epithelial markers ZO1 in cervical cancer cells decreased significantly,and the interstitial marker Vimentin increased significantly.4.The results of mRNA microarray showed that the differentially expressed genes in the silencing group and the control group were highly enriched with mTOR,TGF-β,and homologous recombination pathways in HeLa cells.Further screening is under way.ConclusionKNTC1 is highly expressed in cervical cancer tissues and cell lines,and its abnormally high expression inhibits the occurrence and development of cervical cancer.The impaired function of KNTC1 promotes the proliferation of cervical cancer cells and enhances the migration and invasion of cervical cancer cells by promoting EMT.