Studies Of SIRT3 Influences The Invasion Of Cervical Cancer Cells By Regulating The Key Enzymes Of Fatty Acid Metabolism

Objective:This study was to investigate the effect of Sirtuin protein 3(SIRT3)on fatty acid metabolism in cervical cancer and its mechanism.Method:(1)To detect the transcription level of SIRT3 in cervical squamous cell carcinoma(CSCC),cervical intraepithelial neoplasia(CIN),chronic cervicitis by qRT-PCR;(2)To detect SIRT3 protein in the above three groups And expression levels in lymph node metastasis tissues through immunohistochemical method.(3)Lentiviral transfection technique up-regulates the expression of SIRT3 in cervical cancer SiHa and C33a cells,and verifies the transfection efficiency by qRT-PCR and Western blotting;acetyl-CoA carboxylase is detected by immunoprecipitation and Western blotting.Protein and acetylation levels of acetyl-CoA carboxylation(ACC1)and Carnitine palmitoyltransferase(CPT1?).(4)Cell healing test,Transwell and nude mouse xenograft modeling to detect the effect of SIRT3 expression changes on tumor cell migration,invasion,proliferation and metastasis.Result:(1)The mRNA expression of SIRT3 in CIN and CSCC tissues was significantly higher than that in chronic cervicitis group(P<0.001).(2)The protein expression of SIRT3 showed an increasing trend in chronic cervicitis,CIN and CSCC tissues,especially in tumor tissues with lymph node metastasis(?~2=56.63,P<0.001).The change of protein expression level was positively correlated with the pathological grade of cervical cancer(?~2=5.776,P=0.039),clinical stage(?~2=11.897,P=0.002)and lymph node metastasis(?~2=12.303,P=0.002).(3)After up-regulating the expression of SIRT3,the ACC1 and CPT1?acetylation levels were decreased,ACC1 protein was decreased and CPT1?protein expression was decreased,and the lipid content in SiHa cells was promoted(P<0.001).After SIRT3 expression was down-regulated,ACC1 and ACC1 were enhanced.The level of acetylation of CPT1?decreased the lipid content in C33a cells(P<0.001).(4)Increased expression of SIRT3 promoted migration,invasion,proliferation and metastasis of cervical cancer cells(P<0.001);decreased expression of SIRT3 and inhibited migration,invasion,proliferation and metastasis of cancer cells(P<0.001).Conclusion:(1)The expression of SIRT3 mRNA and protein in cervical cancer tissues is significantly increased,which is associated with poor prognosis parameters.(2)SIRT3 is highly expressed in cervical cancer cells,which can reduce the acetylation level of ACC1 and CPT1?,enhance fatty acid synthesis,inhibit the?oxidation of fatty acid,increase the lipid content in tumor cells,promote the proliferation and migration of tumor cells,and potentially potential for cervical cancer,to be the experimental basis for the potential therapeutic target research of cervical cancer.