The Mechanism Of Hsa＿circ＿0008621 Regulating EMT In Salivary Adenoid Cystic Carcinoma

Objective:Salivary adenoid cystic carcinoma(SACC)of the salivary gland is one of the common malignant tumors in the head and neck,which is prone to the invasion and metastasis of autophagic nerve.Surgical treatment is mainly adopted in clinical practice.However,postoperative recurrence and distant metastasis are still the main causes of death for patients with SACC.According to the pathogenesis of cancer,in the process of early cancer formation,tumor cells are in an epithelial-like state.With the development of tumor,cells with mesenchymal characteristics are gradually formed.Epithelial-mesenchymal transition(EMT)is considered to be an important part of the deterioration of various tumors.Tumor invasion and metastasis are closely related to epithelial-mesenchymal transition(EMT),and are the main causes of SACC metastasis.Circ RNA(mi RNA)is another endogenous non-coding RNA discovered after micro-RNA and long-chain non-coding RNA,which can regulate and drive tumor genes and immunotherapy targets.Circ RNA has been confirmed to play an important role in the occurrence,development of tumors and immunotherapy.However,it is not clear whether hsa＿circ＿0008621 can affect the occurrence and development of SACC through EMT.Therefore,the main purpose of this study is to explore the potential regulatory mechanism of hsa-circ-0008621 in EMT of salivary adenoid cystic carcinoma,so as to provide new targets for clinical diagnosis and treatments.Methods: 1.The expression levels of EMT markers Vimentin,E-cadherin,Snail,Slug,Zeb1,and Twist in 20 SACC clinical tissue samples were detected by immunohistochemistry;The expression of EMT markers in tumor tissues and normal tissues was detected by Western blot.2.Screen the ideal differentially expressed circRNA in clinical tissue samples of SACC by qRT-PCR.3.Use small interfering RNA technology to silence the hsa＿circ＿0008621 and interfere with its expression in SACC-83 and SACC-LM cells.qRT-PCR was used to detect the transfection rate.Western blot was used to detect the expression of EMT markers,effect on SACC-83 and SACC-LM cell migration and invasion ability.4.Transwell cell invasion and migration assay was used to detect the changes in invasion and migration ability of two tumor cell lines before and after interference with circRNA.5.Comparison of animal experiments with viruses that interfered with hsa＿circ＿0008621 in SACC-LM cells to form a stable expression cell line.SACC-LM and SACC-LM that interfered with hsa＿circ＿0008621 were implanted into the submandibular tumors of nude mice.The expression of EMT markers E-cadherin and Vimentin were observedResults: 1.Results of immunohistochemical experiments showed that EMT-related transcription factors were positively expressed in SACC tissues.Western blot showed that compared with the normal glandular gland of the control group,the expression of the epithelial cell marker E-cadherin decreased,the expression of the mesenchymal cell marker Vimentin increased,and the expression of EMT-related transcription factors Snail,Slug,twist,zeb1 increased.2.The expression of hsa＿circ＿0008621 in SACC was detected by qRT-PCR,and it was found to be highly expressed in SACC.3.Culture SACC-83 and SACC-LM,and use small interfering RNA of hsa＿circ＿0008621 to interfere with their expression in both cells.qRT-PCR detected low expression after transfection.Western blot detection of EMT marker expression showed that E-cadherin expression increased,vimentin expression decreased,and EMT-related transcription factor Zeb-1 expression decreased.4.Transwell and other methods found that the invasion and migration ability of the two cells were reduced.5.Tumor growth was inhibited in si＿circ＿0008621 group compared with control.Conclusions: Hsa＿circ＿0008621 promotes migration,invasion of SACC cells by regulating EMT process.