Expression Of DAB2IP In Cutaneous Squamous Carcinoma And Basal Cellcarcinoma And Its Mechanism Of Invasion Of Squamous Cellcarcinoma

BackgroundNonmelanoma skin cancer(NMSC)is composed of Basal Cell Carcinoma(BCC)and Squamous Cell Carcinoma(SCC).Cutaneous Squamous Cell Carcinoma(cSCC)is originated in the skin or adnexal cutin to form a kind of malignant tumor cells,accounting for about 20%-30%of all non melanoma skin cancer,which is the second most common keratinocyte cance[1-3].cSCC has an estimated incidenceof700,000casesintheUnitedStatesperyear and has displayed a sharp rise in incidence during the last 2 decades[4,5].In contrast to BCC,which has high capacity for local destruction but low risk for metastasis,cSCC is more aggressive with increased potential for regional and distant spread[6,8].Although most cases of cSCC have an excellent prognosis after surgical removal,there are particular high-risk features that are predictive of more aggressive clinical behavior[1].These features include tumor diameter,poorly or undifferentiated histology,presence of perineural invasion,invasion beyond subcutaneous fat,and certain anatomical locations.The single most important prognostic indicator for mortality in patients with SCC is the presence or development of regional lymph node metastasis[9.10].It has been reported that 3.7%to 5.2%of cSCC patients develop nodal metastasis(NM),and as a result 1.5%to 2.1%suffer disease-spific death[6.9-13].Furthermore,althoughthefatalityratefor SCC is only approximately 1%,the overall mortality figures equal or exceed those for melanoma,which is far more lethal,but less common[14].Bowen's Disease(BD)is a relatively common in situ Squamous cell carcinoma originating from epidermal la?? ated Squamous cells(SCCIS).Although the tumor cells were only confined to the epidermis and did not break through the basement membrane to immerse in the dermis,they also had a certain infiltration and invasion potentia[15].DAB2 interacting protein(DOC-2/DAB2 interactive protein,DAB2IP)is the recent discovery of the Ras-guanosine triphosphatase activator protein(Ras GTPase activating protein,Ras-GAP),a new members of the Ras-GAP family,located on chromosome 9 q33.1-q33.3.it has biological functions of regulating cell apoptosis,inhibiting angiogenesis,inhibiting tumor growth,metastasis and radiotherapy tolerance[16].DAB2IP was first found to be down-regulated in lung cancer[17].So far,prostate cancer and liver cancer have been relatively clear in the studies on the function and molecular mechanism of DAB2IP[18].It is believed that DAB2IP may be a tumor suppressor gene and has certain value for the prognosis of tumors.It has been found that the expression of Matrix MetalloProteinase(MMP-2)and Vascular Endothelial Growth Factor(VEGF)in cSCC is related to the degree of differentiation and lymph node metastasis.The expression of Nuelear Factor-B(NF-B)and Tissue Inhibitor MetalloProteinase(TIMP-2)in cSCC was related to the degree of differentiation and lymph node metastasis.Poorly differentiated squamous cell carcinoma can produce more MMP-2 to degrade the Extracellular Matrix(ECM),thus contributing to local invasion of the tumor.MMP-2 and VEGF may play a synergistic role in the invasion and metastasis of cSCC.Expression of Transforming Growth Factor(TGF-1)was independent of tumor differentiation,invasion,and lymph node metastasis.On the basis of previous studies,the expression of DAB2IP gene and protein in cSCC and Bowen's Disease was further discussed,and the relationship between them and patients,gender,age,site of onset,skin lesion size,skin lesion type,lymph node metastasis,pathological manifestations and recurrence was analyzed.The biological function of DAB2IP was further studied by in vitro experiaents.ObjectivePathological and clinical data of patients with cSCC and Bowen,s Disease were collected.The correlation analysis method was used to explore the relationship between DAB2IP gene and protein expression and clinical phenotype of patients with cSCC.The significance and value of DAB2IP in cSCC and Bowen's Disease were evaluated and clarified.Though cell culture in vitro,infection target cells by lentivirus vectors,and PCR technology,we explore the pathogenesis of cSCC by building DAB2IP-knockdown cell model,further clarify the effects of DAB2IP expression on the function and behavior of cSCC cells,providing a theoretical and practical basis for the formulation and promotion of clinical targeted therapy.Methods1)The general data and clinical features of cSCC,bowen's disease and basal cell carcinoma,as well as the recurrence of follow-up patients were analyzed retrospectively.2)The FQ-PCR and inmunohistochemical method were used respectively to detect the expression of DAB2IP gene and protein level in cSCC of the cutaneous and Baowen'S disease and skin BCC,analysis the correlation between the expression patterns of DAB2IP gene and the clinical phenotype in cSCC and Bowen's Disease and BCC tissue.The level of DAB2IP gene was detected by RT.PCR in vitro model cells.Construction of a cell model with DAB2IP knockout using RNAi technology,and then through the cell scratch experiment,Transwell experiment,gelatin enzyme spectrum experiment and CCK 8 proliferation,plate clone formation experiment,DAPI staining and Hoechst 33258 staining,cell proliferation and apoptosis induction ability were detected,and the role of DAB2IP in cSCC was observed and discussed.Results1.The expression of DAB2IP protein:in cutaneous squamous cell carcinoma was significantly higher than that in benign dermal vegetation,with a positive rate of 42.3%in benign dermal vegetation and 85.4%in cSCC,showing a statistically significant difference.2.Among the 48 patients with cSCC,34(70.8%)showed high exPression of DAB2IP protein(++?+++),and 14(29.2%)showed low expression(0?+).The expression of DAB2IP protein was positively correlated with the age,depth of infiltration(whether to break through the basement membrane)and clinical stage of patients with cSCC,(x2=4.55,4.38,10.76,respectively,P<0.05).There was no significant correlation with gender,tumor size,tumor location and clinical phenotype,x2=0.13,2.96,1.32,0.01,respectively,P>0.05)and the difference was statistically significant.3.The expression abundance of DAB2IP in SCL-1 cells was medium.4.After infection of scl-1 cells by lentiviral vector,compared with the control group,the proliferation of cells was slowed down.The number of cell clones decreased.There were fewer cells in S phase and more cells in G1 phase and G2/M phase.The number of apoptosis increased.The ability of cell metastasis,migration and invasion were decreased,and the difference was statistically significant.5.The expression of DAB2IP showed weak positive or no expression in most benign dermatitis groups(control group),and a few showed high expression.In the basal cell carcinoma group(case group),33 cases showed strong positive expression of DAB2IP.The positive expression rates of DAB2IP protein in the control group and case group were 42.3%and 93.7%,respectively,(x2?15.00,p<0.01),with statistically significant differences.6.There was no statistically significant difference in the positive rate of the expression of DAB2IP protein in basal cell carcinoma between different age groups,gender groups and large tumor groups,(x2?1.54,1.91,0.22 respectively,P>0.05),but the difference in superficial(carcinoma in situ)group was significantly lower than that in invasive group,(x2?6.47,P<0.05),with statistically significant difference.Conclusion1.the increased expression of DAB2IP in cSCC may be related to the occurrence and infiltration of cSCC.2.Knockdown the expression of DAB2IP inhibited proliferation and invasion of SCL-1 cells in vitro.These fingdings:indicated that high expression of DAB2IP contribute to tumor development and proliferation in cSCC.3.the increased expression of DAB2IP in basal cell carcinoma of the skin may be related to the occurrence and infiltration of basal cell carcinoma of the skin.