多聚腺苷酸结合相互作用蛋白1在乳腺癌组织中的表达及其对乳腺癌细胞增殖、迁移和糖酵解的影响作用研究OA

Background Current studies indicate that poly A‑binding‑protein‑interacting protein 1(PAIP1)is highly expressed in various tumors,yet research on its role in breast cancer remains scarce,and the underlying mechanisms are not yet fully understood.Objective To investigate the expression of PAIP1 protein in breast cancer and its relationship with clinicopathological characteristics,and to explore the role of PAIP1 in the biological functions of breast cancer cell proliferation,invasion and glycolysis.Methods Collect data from the GEO database(841 cases of breast cancer tissue and 631 cases of adjacent tissue)and the TCGA database(1 060 cases of breast cancer tissue and 111 cases of adjacent tissue,including different clinical stages of breast cancer:stage Ⅰ,351 cases;stage Ⅱ,379 cases;stage Ⅲ,364 cases;stage Ⅳ,77 cases)to analyze PAIP1 mRNA expression.Enroll 159 patients with invasive breast cancer who were treated in the Breast Surgery Department of Yanbian University Hospital from February 2012 to March 2013 as the study subjects.Collect surgical specimens of cancerous and adjacent tissues,and detect PAIP1 expression using immunohistochemistry.Transfect small hairpin RNA(sh-RNA)to silence PAIP1 expression in two breast cancer cell lines,SK-BR-3 and MCF-7,and divide the cells into three groups:the Control group,the PAIP1-shRNA group,and the PI3K activator(740Y-P)+PAIP1-shRNA group.Use the cell counting kit-8(CCK8)method and plate cloning experiments to assess the proliferative capacity of the two breast cancer cell lines.Conduct scratch assays and cell migration experiments to evaluate the migration ability of the two cell lines.Employ Western blotting to examine the expression of proteins related to the phosphoinositide 3-kinase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR)signaling pathway,as well as E-cadherin and vimentin.Measure ATP,lactate,and glucose levels in the two breast cancer cell lines using colorimetric methods.Results According to the immune histochemical analysis results from the GEO and TCGA databases and clinical collected case specimens,the PAIP1 mRNA expression levels in adjacent tissues were consistently lower than those in breast cancer tissues(P<0.05).Additionally,TCGA data analysis revealed that PAIP1 mRNA expression levels in clinical stage Ⅱwere lower than those in stages Ⅲ-Ⅳ(P<0.05).CCK8 and colony formation assays demonstrated that in both breast cancer cell lines,the PAIP1-shRNA group exhibited lower proliferation capacity and colony formation counts compared to the Control group(P<0.05),while the 740Y-P+PAIP1-shRNA group showed higher values than the PAIP1-shRNA group(P<0.05).Scratch and cell migration assays indicated that in both breast cancer cell lines,the PAIP1-shRNA and 740Y-P+PAIP1-shRNA groups displayed reduced lateral and longitudinal migration capabilities compared to the Control group(P<0.05),with the 740Y-P+PAIP1-shRNA group showing higher migration than the PAIP1-shRNA group(P<0.05).Western blot results revealed that in both breast cancer cell lines,the PAIP1-shRNA group exhibited lower protein expression levels of phosphorylated PI3K(p-PI3K),phosphorylated AKT(p-AKT),phosphorylated mTOR(p-mTOR),Vimentin,hexokinase 2(HK2),glucose transporter 2(GLUT2),glucose transporter 3(GLUT3),and lactate dehydrogenase(LDH)compared to the Control group(P<0.05),while E-Cadherin levels were higher(P<0.05).The 740Y-P+PAIP1-shRNA group showed higher protein expression levels of p-PI3K,p-AKT,p-mTOR,Vimentin,HK2,GLUT2,GLUT3,and LDH than the PAIP1-shRNA group(P<0.05),but lower E-Cadherin levels(P<0.01).Additionally,HK2,GLUT2,GLUT3,and LDH expression levels were lower than those in the Control group(P<0.05).Glucose,lactate,and ATP detection results showed that in both breast cancer cell lines,the PAIP1-shRNA and 740Y-P+PAIP1-shRNA groups exhibited lower ATP,glucose,and lactate levels than the Control group(P<0.05),with the 740Y-P+PAIP 1-shRNA group showing higher levels than the PAIP1-shRNA group(P<0.01).Conclusion PAIP1 protein is highly expressed in breast cancer tissue.Silencing PAIP1 inhibits the proliferation,migration and glycolysis of breast cancer cells via inhibiting the PI3K/AKT/mTOR signaling pathway.