The Role Of Notch Signaling Pathway In Migrate And Epithelial-mesenchymal Transition Of Cervical Cancer Cell

Cervical cancer is one of the most common gynecologic malignancies and the second most common cancer in women.More than 500,000 new cases are reported each year,and most cases occur in developing countries,especially in these rural areas.Among them,Central America and South America,the Caribbean,sub-Saharan Africa and South Asia are the areas with the highest incidence of cervical cancer,and female patients are often between the ages of 15 and 44.In some areas,cervical cancer has even surpassed the first breast cancer in women with malignant tumors.In recent years,with the development and promotion of early screening,detection techniques and treatment methods,the diagnosis and treatment of cervical cancer has made remarkable progress,but due to geographical,economic and social and cultural barriers and other reasons,Early screening is limited in many developing countries.In India,the diagnosis of cervical cancer is usually late,with more than 80%of Indian women suffering from locally advanced(IIB-IVa),and less than 50%in industrialized countries.The prognosis of early cervical cancer is generally good;however,the 5-year survival rate of locally advanced cervical cancer is gradually reduced,from 58%in stage IIB,35%in stage IIIA,32%in stage IIIB,and even 16%in stage IVa.Overall,the prognosis of cervical cancer is extremely poor,and the 1-year survival rate of patients with recurrent or advanced cervical cancer is only 10%-20%.Cervical cancer treatment is based on the clinical stage of the International Federation of Obstetricians and Gynecologists(FIGO),which includes surgery,radiation,and chemotherapy alone or in combination.These treatments are generally effective,and residual tumors are still observed in 40-50%of patients with advanced cancer.The occurrence of chemotherapy drug tolerance,tumor invasion and metastasis leads to treatment failure and is an important cause of death in most patients.Human papillary virus(HPV)is a member of the family Papilloviridae,which has a non-encapsulated double-stranded circular dna(dscdna)genome with approximately 8kb.It is highly predisposed to skin and mucosal epithelial cells and can lead to various tumors,including vulva,vagina,penis,anus,esophagus.Cancer of multiple organs,such as the head and neck.Up to now,there are more than 100 kinds of hpv,which can be classified into skin type and mucosal type according to tissue tendency,high risk type(HR-HPV)and low risk type(LR-HPV)according to carcinogenic potential.Worldwide,viral infections account for about 15-20 percent of all human cancers,of which 5 percent are caused by hpv infections.Numerous epidemiological,clinical and experimental evidence studies have found that cervical cancer is closely related to HPV infection,especially high-risk human papillomavirus(HR-HPV),which occupies an very important position in cervical carcinogenesis,including 13 Types of HR-HPV(HPV types 16,18,31,33,35,39,et al)are thought to be associated with precancerous cervical lesions and invasive cervical cancer,16 and Type 18 HPV is the most common and highly carcinogenic type that can cause 70%to 80%of cervical cancer.On the other hand,there are 12 different LR-HPV types(6,11,40,42,43,44,53,54,61,72,73 and 81)associated with genital warts and benign cervical lesions,such as sharp Wet warts,etc.,in which HPV 6 and 11 mainly cause more than 90%of genital warts.In addition,early sexual exposure,multiple sexual partners,smoking,long-term oral use of contraceptive and co-infection with herpes simplex virus 2 or human immunodeficiency virus(hiv)may increase the risk of the disease.The oncoprotein in HPV can maintain cell proliferation signals,evade the regulation of growth inhibitory factors,continuously replicate cells,resist apoptosis,sustain angiogenesis,invade and metastasize,lead to the development of precancerous lesions,and eventually develop into invasive cancer.These processes are regulated by a variety of signaling pathways,and work together to regulate the occurrence and development of tumors.Therefore,the role of tumor-associated signaling pathways in cervical cancer cells has gradually attracted the attention of scholars.These altered signaling pathways play a crucial role in the initiation and maintenance of HPV-associated cancers.The signaling pathway provides a targeted treatment for cervical cancer.Current research indicates that persistent "high-risk" human papillomavirus(HR-HPV)infection is a prerequisite for cervical cancer,and HPV infection is found in up to 95%of diagnosed cervical cancer cases.The early(E)region encoding proteins of HPV include E1,E2,E4,E5,E6 and E7,which are required for viral replication and transcription;the late(L)region encodes viral assembly and requires two structural capsid proteins,L1 and L2.In the process of HPV-related carcinogenesis,over-expressed E6 and E7 oncoproteins are the main pro-cancer factors for cervical epithelial cells to transform into tumors,maintaining the transformed phenotype of cancer cells,thus epithelial-mesenchymal transition(EMT)during cervical carcinogenesis.)participated in it and played an important role.EMT refers to the physiological process of obtaining the biological characteristics of some mesenchymal cells from differentiated and mature epithelial cells,which are closely related in tumorigenesis,development and tumor invasion,metastasis and recurrence.Most of human cancers are associated with mouse solid tumor by epithelial solid tumor signal.The(mmtv)integration test of mouse mammary tumor virus is the earliest.Decades ago,intracellular notch 1 was detected for the first time in human cervical cancer,prompting research into the possible role of this pathway in cancer.However,there is a complex interaction between notch signal and hpv.Some experiments have reported that the complex interaction of their transcriptional regulation has dose-dependent effect,and some data have proved that the deregulated notch signal can promote epithelial transformation.In the process of carcinogenesis associated with hpv,many signal pathways and protein molecules play a regulatory role in the carcinogenesis and metastasis of cervical cancer cells,in which emt enhances the invasion of cervical cancer cells.Metastasis plays a key role.EMT not only regulates the occurrence and development of various tumor cells,but also is regulated by complex signaling pathways and molecules involved in the maintenance and regulation of emt.However,the function of notch signaling pathway in the biological process of cervical cancer emt is unclear.Therefore,by studying the role and mechanism of notch signaling pathway in the formation of cervical cancer cells,drug resistance and emt transformation,specific targeted therapy was carried out on the biological processes such as proliferation and drug resistance of cervical cancer cells.It will be of great significance to improve the therapeutic effect of cervical cancer.In this study,we confirmed the effect of notch signaling pathway on the proliferation,drug resistance and emt of cervical cancer cells.We found that notch signaling pathway plays an important role in the regulation of cervical cancer proliferation and emt.The specific inhibition of notch signaling pathway can effectively inhibit the proliferation,migration and tumorigenesis of cervical cancer cells,and provide a reliable experimental basis for the combination and targeted therapy of cervical cancer.Part-I The role of Notch signaling pathway in cervical cancer cellsObjectives:The regulatory mechanism of notch signaling pathway on tumor genesis and development is very complex,but in the field of cervical cancer,the specific mechanism of notch signaling pathway has not been clarified.In this paper,we will examine the expression of notch signaling pathway and downstream molecules in cervical cancer cells,and study the effect of notch signaling pathway on the proliferation,drug resistance and metastasis of cervical cancer cells.To provide experimental evidence for clarifying the mechanism of notch signaling pathway in cervical cancer cells.Methods:Flow cytometry was used to detect the expression of notch gene in hpv-16-immortalized cervical epithelial cell line and cervical cancer cell line.The transcriptional and expression differences of notch signaling pathway related receptor notchl and its downstream target gene hes-1 in the two groups were detected.Plate clone formation assay and serum-free suspension culture were used to evaluate the effect of notch signal pathway inhibitor on cervical cancer cell tumorigenesis in vitro.The effect of drug resistance.Results:1.Notch2 mRNA and protein expression was highest in the metastatic cervical cancer cell line Caski cells,and second in the non-metastatic cervical cancer cell line Hela cells,and lowest in the normal cervical epithelial cell line CRL2614 cells;2.Notch signaling pathway inhibitor(RO4929097)can significantly reduce the proliferation,migration and invasion of cervical cancer cell lines;3.The sensitivity of RO4929097-treated cervical cancer cells to cisplatin-induced cytotoxicity is increased.Conclusions:Notch signaling pathway and target genes are up-regulated in cervical cancer cell lines,and its signaling pathway inhibitor(RO4929097)blocks Notch pathway and inhibits proliferation,migration and invasion of cervical cancer cell lines,and inhibits Notch signaling pathway to increase cervical cancer The sensitivity of cells to chemotherapeutic drugs plays an important role in the occurrence and metastasis of cervical cancer.Part-II The role of Notch signaling pathway in cervical cancer cell line EMTObjectives:The activation of emt is a key step for tumor cells to obtain invasion and metastasis.As one of the signal regulatory pathways in the process of emt,notch signaling pathway also plays an important role in the process of tumor resistance and metastasis.But in cervical cancer cells,there is no literature to confirm whether there is emt transformation in cervical cancer cells,and it is not clear how the notch signaling pathway regulates the emt of cervical cancer cells.In this chapter,we will explore the expression of emt in cervical cancer cells,verify the mechanism of notch signaling pathway on emt,and provide a new idea for the targeted therapy and combined drug therapy of cervical cancer.Methods:Western blotting was used to detect the transcription level of emt markers in cervical cancer cells,and the migration and infiltration ability of low cervical cancer cells was detected by cell scratch repair test and transwell cell migration assay.The effect of notch signaling pathway inhibitor on emt in cervical cancer cells was detected.Results:1.Inhibition of notch signal transduction pathway can reduce the expression of emt specific markers in cervical cancer cell line.2 the inhibitor of notch signal pathway can reduce the migration and invasion ability of cervical cancer cells.Conclusions:Cervical cancer cells undergo emt transformation and notch signaling pathway plays a synergistic role in the process of cervical cancer invasion and metastasis,and plays a synergistic role in the development of emt in cervical cancer cells.