Mechanism Of LASP1 Interacting Protein N-WASP Inducing Tumor Budding And Promoting Colorectal Cancer The Invasion And Metastasis

PurposeColorectal cancer(CRC)is one of the common malignant tumors in clinical diagnosis,because of its high morbidity and mortality,which has threatened human health[1].the mechanism of invasion and metastasis involves multiple stages and genes.It is a complex biological process.Tumor sprouting as an important morphological feature of malignant tumors,and its degree of sprouting can indirectly prompt the invasion and metastasis of colorectal cancer,and is related to poor prognosis[2].Studies have shown that the invasion and metastasis of cancer cells depend on the formation of invasive pseudofoot(Invadopodia)[3]and the actin assembly under the cell membrane regulates this process;so far,many genes have been involved,and ARP2/3 complex Catalysis is an important key link[4]and research has shown that it can be activated by the WASP(Wiskott-Ardrich Syndrome Protein)family[5].LASP1(LIM and SH3 protein 1)is a colorectal cancer-related metastasis protein studied by our research group[6].Previous studies have found that LASP1 can bind and activate the WASP(Wiskott-Ardrich syndrome protein)family.This study takes the high expression of WASP(Wiskott-Ardrich Syndrome Protein)family in colorectal cancer tumor tissues as the entry point,and combines clinical investigation with basic experiments to reveal the important role of WASP in the invasion and metastasis of colorectal cancer.Clinical prognosis and treatment of tumors provide new biological targets.Methods1.Immunohistochemistry(IHC)to study the expression of WASP and N-WASP in colorectal cancer tissues;the effect of the expression of N-WASP on the microenvironment of colorectal cancer and the correlation of aggressive frontiers;Expression and clinical prognosis in colorectal cancer;2.Construct stable N-WASP,LASP1 overexpression and N-WASP interference cell line RKO,Western Blot detects its expression and correlation;3.Nude mice tumor formation and abdominal mesenteric planting in situ;explotion of invasion and metastasis;4.Co-immunoprecipitation method to verify the interaction and domain between N-WASP and LASP1;5.Co-IP and immunofluorescence(IF)verify that LASP1 and SH3(LASP1 domain)promote the binding of N-WASP and ARP3.Results1.IHC experiments found that colorectal cancer tissues were low or even non-expressing WASP,and N-WASP was highly expressed;N-WASP was highly expressed in colorectal cancer tissues;which expression is related with lymph node metastasis,inflammatory response,tumor invasion pattern and tumor bud grade-;2.In situ implantation experiments of subcutaneous tumors and abdominal mesentery in nude mice,N-WASP promotes the invasion and metastasis of colorectal cancer;3.CO-IP experiment,LASP1 binds to N-WASP through its SH3 domain;4.In the CO-IP and IF experiments,compared with the control group,overexpression of LASP1 or SH3 promoted the combination of N-WASP and ARP3,and the strength was sequentially strong,medium,and weak;5.IHC proved that the expression of N-WASP and LASP1 was associated with lymph node metastasis;ARP3 was highly expressed in tumor tissues;the expression of N-WASP,LASP1 and ARP3 in colorectal cancer tissues was positively correlatedConclusion1.N-WASP is highly expressed in colorectal cancer tumour tissues;2.High expression of N-WASP is associated with microenvironment and tumour budding of colorectal cancer;3.N-WASP promotes the invasion and metastasis of colorectal cancer;4.N-WASP promotes the binding of N-WASP and ARP3 by binding to the SH3 domain of LASP1;5.the expression of N-WASP and LASP1 is related to clinical prognosis,and the expression of N-WASP,LASP1 and ARP3 is positively correlated.Innovative1.Basically connect with the clinic to verify the role and potential mechanism of N-WASP in the invasion and metastasis of colorectal cancer;2.It is suggested that the high expression of N-WASP in colorectal cancer is associated with tumor sprouting.