Effect Of CNTN1 Gene Silencing On Metastasis Of Canine Mammary Gland Tumer Cell Line

CNTN1 gene is a newly discovered cancer-related gene in recent years.Early studies have shown that CNTN1 gene is involved in the development and progression of synaptogenesis,and until 2006,CNNT1 c DNA whole genome sequencing was used by Su et al.for the first time,found which was associated with invasion and metastasis of lung cancer,so the role of CNTN1 gene in the tumor was the researchers attention,the mechanism of CNTN1 gene in tumor is still unclear until now.Therefore,canine mammary gland tumor cell CHMm was used as an experimental material to study the mechanism of CNTN1 gene on metastasis and invasion in canine mammary gland tumorThe design and synthesis of CNTN1 sh RNA plasmid was commissioned by Gemma,and which was transfected into the mammary gland tumor cell line CHMm cells by using liposome 2000 transient transfection method.The cell proliferation was detected by CCK-8 and cell plate clone assay,apoptosis was detected by Annexin V-ACP/7-AAD apoptotic kit,cell migration ability was detected by cell wound scratches.and western blot was used to detect the changes of Akt signaling pathway proteins to investigate the effect of CNTN1 gene silencing on CHMm cells and Akt signaling pathway in breast cancer.Result:1.A large number of green fluorescence was observed in CHMm cells transfected with CNTN1 sh RNA plasmid under fluorescence microscopy,the expression of CNTN1 protein in CHMm cells transfected with CNTN1 gene was significantly lower than that in the control group analyzed by Western blot(p<0.05).2.Proliferation and apoptosis of canine breast cancer cell lines was not affected by transfection of CNTN1 sh RNA plasmid(p>0.05).3.The expression of PI3K、Akt(Ser473)and mTOR protein in the cell group transfected with CNTN1 sh RNA plasmid was significantly lower than that in the control group(p<0.05).However,the ratio of m TOR/p-m TOR protein expression was not significantly different from that of the control group(p>0.05).4.The expression of Bax,caspase-3 and caspase-9 protein in the CNTN1 gene group was significantly higher than that in the control group(p<0.05).The expression of Bcl-2 protein was significantly lower than that of the control group(p<0.05).5.The expression of Cyclin D1 protein in the cell group transfected with CNTN1 gene was significantly lower than that in control group(p<0.05).The expression of P21 and P27 kip protein was significantly higher than that of control group(p<0.05).6.The expression of e NOS protein was significantly lower than that of the control group(p<0.05).The expression of E-cadherin protein was significantly higher than that of the control group(p <0.05).CNTN1 gene was successfully silenced in CHMm cell,and the silencing efficiency could reach 60%-70%.Cell proliferation was not attected by CNTN1 silencing,Akt signaling pathway could be regulated by CNTN1 silencing,which regulates the expression of downstream proteins in this pathway,providing a new target for breast cancer-based therapy.