Preliminary Study On The Mechanism Of Inhibiting Element L Silencing Transcription Factor(REST) In The Reg?lation Of KCa3.1 Expression In Cervical Squamous Cell Carcinoma

Objective:Conductance in calcium activated potassium channels(KCa3.1)expression in cervical tissue and the occurrence and development of cervical squamous cell carcinoma has a close relationship,discusses component l silence transcription factor(REST)is also called silence neurons restrictive factor(neuron restrictive silencing factor,NRSF)expression in cervical squamous cell carcinoma tissue and the conductivity of calcium activated potassium channels(KCa3.1),the influence of the initial study KCa3.1 raised in cervical squamous cell carcinoma of the mechanism.Methods:By cons?lting and organizing databases(NCBI,UCSC,Bio GPS),the expression of REST in different tissues was understood,and the expression of REST in different tissues and cervical tissues was analyzed.REST and KCa3.1mRNA expression levels in different tissues were detected by real-time fluorescence quantitative PCR(qrt-pcr),and REST protein expression levels in different tissues were detected by immunohistochemical methods.Chromatin immunoprecipitation(ChIP)and quantitative PCR assays were used to determine whether REST can be enriched at the KCa3.1 promoter inhibitory element 1(RE1)site and to what extent.Res?lts: Through analysis and collation of the database,it is found that REST is highly expressed in normal non-neuraltissues and highly expressed in normal cervical tissues.The relative expression levels of RESTmRNA in cervical squamous cell carcinoma,adjacent carcinoma and normal cervical tissue were 0.46 0.06,0.89 0.13 and 1.00 0.12,respectively.The relative expression of RESTmRNA in cervical squamous cell carcinoma decreased by 48% and 54%,respectively,compared with that in paracancerous tissue and normal cervical tissue.The expressions of KCa3.1mRNA in cervical squamous cell carcinoma,paracancerous tissue and normal cervical tissue were 2.14 0.35,1.18 0.21 and 1.00 0.13,respectively.KCa3.1mRNA expression in cervical squamous cell carcinoma increased by 81% and 114%,respectively,compared with that in paracancerous tissue and normal cervical tissue.RESTmRNA expression in cervical squamous cell carcinoma was significantly lower than that in adjacent and normal cervical tissues(P < 0.05),and KCa3.1mRNA expression in cervical squamous cell carcinoma was significantly higher than that in adjacent and normal cervical tissues(P < 0.05).Immunohistochemical found in the cytoplasm and nucleus can REST protein positive reaction was observed,and the REST were detected in normal cervical tissue protein expression of different level,the positive rate was 100%,the REST in cervical squamous cell carcinoma tissue protein positive rate was 40%,the REST quantity of protein expression in cervical squamous cell carcinoma tissues from normal cervical tissue,significantly decreased(P < 0.05).Chip-qpcr showed that REST co?ld bind at RE1 site in the KCa3.1 promoter region,and the enrichment of REST at RE1 site in normal cervical tissue,paracancer tissue and cervical squamous cell carcinoma tissuewas 0.41 0.03,0.36 0.041 and 0.16 0.008,respectively.REST concentration in cervical squamous cell carcinoma decreased by about 60.1% compared to normal cervical tissue and 56% compared to paracancer tissue.REST concentration in cervical squamous cell carcinoma was significantly lower than that in normal cervical tissue and paracancer tissue(P<0.05).Conclusion:REST expression in cervical squamous cell carcinoma was significantly lower than that in normal cervical tissue and adjacent tissue,and REST co?ld negatively reg?late the expression of KCa3.1 by binding to the KCa3.1 promoter RE1 site.Early res?lts show KCa3.1 increased expression in cervical cancer and promote the growth of cervical cancer cells,increase the ability of invasion and metastasis of cervical cancer cells,so the cervical squamous cell carcinoma tissue REST cut lead to its inhibition of the expression of KCa3.1 gene expression is abate,lead to increased KCa3.1 gene expression and enhance KCa3.1 activity of potassium channels,promote cervical squamous cell cancer cell growth,invasion and metastasis.