The Study Of The Effect Of NAC1 On Proliferation,migration And Invasion Of Human Cervical Cancer HeLa Cell And Its Mechanisms

Objective:The nucleus accumbens-associated protein-1(NAC-1),a nuclear transcription coregu Lator belonging to the BTB/POZ gene family,is encoded by NACC1 gene.The N-terminal of NAC1 contains the BTB domain,which can mediate its binding with other molecu Les to form a NAC1 transcription complex,which is involved in the regu Lation of a variety of biological functions and is closely related to the proliferation,migration and invasion of a variety of tumor cells.However,there are few studies on the relationship between Na C1 and biological behavior of cervical cancer cells.This study aims to study the impact of Na C1 knockdown on proliferation,migration and invasion of cervical cancer HeLa cells,so as to provide an important theoretical basis for targeted therapy of cervical cancer.Method:1.Immunohistochemistry assay was used to determine the protein expression of NAC1 in normal cervical tissues and cervical cancer tissues.2.Four short hairpin RNAs(sh RNA)targeting human NACC1 gene and negative control(NC)were constructed and transfected into HeLa cells to observe the gene silencing effect.RT-q PCR and Western blot were used to screen the plasmids with the highest knockdown efficiency for subsequent experiments.3.CCK8 and clone formation experiments were designed to determine the influence of NAC1 knockdown on the proliferation of cervical cancer HeLa cells.4.Transwell and wound healing were designed to detect the effect of NAC1 knockdown on invasion and migration of HeLa cells.5.Western Blot assay was designed to detect the effects of knockdown of NAC1 on tumor suppressor proteins p53 and p21 of cervical cancer HeLa cells and the changes of EMT-related proteins(E-cadherin,Snail,N-cadherin,Vimentin).To preliminarily explore whether NAC1 promotes the proliferation,invasion and metastasis of cervical cancer cells by regu Lating the mechanisms of p53,p21 and EMT.Resu Lts:1.Immunohistochemistry for NAC1 in some cervical cancer sections produced strong staining of the nuclei,but not in normal cervical tissues.2.RT-q PCR and Western Blot resu Lts showed that NACC1-Sh4 had the highest knockdown efficiency and was used in subsequent cell transfection experiments.3.After NAC1 knockdown,the proliferation and colony forming ability of HeLa cells decreased.4.After NAC1 knockdown,the migration and invasion ability of HeLa cells sepressed.5.After NAC1 knockdown,the expression levels of p53 and p21 elevated,the expression levels of EMT related protein E-cadherin elevated,and the expression levels of N-cadherin,Vimentin and Snail in HeLa cells declined.Conclusion:1.After NAC1 knockdown,the proliferation,colony formation,invasion and migration of HeLa cells decreased significantly.2.After NAC1 knockdown,the expression of p53 and p21 protein in HeLa cells increased,and EMT related molecu Lar indicators changed,suggesting that NAC1 may participate in the occurrence and development of cervical cancer through regu Lating p53,p21 and EMT.