The expressions of Tspan8 and ITGA5 in breast cancer tissues and their relationship with clinicopathological features and prognosis / 国际检验医学杂志

Objective To investigate the expressions of transmembrane protein 8(Tspan8)and integrin α5(ITGA5)in breast cancer and their relationship with clinicopathological features and prognosis.Methods A total of 136 breast cancer patients admitted to the hospital from April 2018 to April 2020 were enrolled in the study.Breast cancer tissue samples and corresponding paracancerous tissue samples were collected.The ex-pressions of Tspan8 and ITGA5 in breast cancer tissues and corresponding adjacent tissues were detected by immunohistochemistry.The relationship between the expressions of Tspan8 and ITGA5 and the clinicopatho-logical characteristics of breast cancer patients was analyzed.The discharged breast cancer patients were fol-lowed up for 36 months,and the survival status of the patients was recorded.The 3-year survival rate of breast cancer patients with different clinicopathological characteristics was compared.Multivariate Cox regression was used to analyze the influencing factors of axillary lymph node metastasis in breast cancer patients.Results The positive expression rates of Tspan8 and ITGA5 in breast cancer tissues were higher than those in adjacent tissues(P＜0.05).The positive expression rates of Tspan8 and ITGA5 in breast cancer patients with poor differentiation,tumor maximum diameter ≥3 cm,TNM stage Ⅲ-Ⅳ,axillary lymph node metastasis and other molecular subtypes were higher than those in moderate/well differentiation,tumor maximum diam-eter＜3 cm,TNM stage Ⅰ-Ⅱ,no axillary lymph node metastasis and triple negative subtype(P＜0.05).The 3-year survival rate of patients with positive expression of Tspan8 and ITGA5 was significantly lower than that of patients with negative expression of Tspan8 and ITGA5(P＜0.05).The 3-year survival rate of patients with poor differentiation,TNM stage Ⅲ-Ⅳ,axillary lymph node metastasis,other molecular sub-types and positive expressions of Tspan8 and ITGA5 were lower than those of patients with moderate/high differentiation,TNM stage Ⅰ-Ⅱ,no axillary lymph node metastasis,triple negative subtype and negative ex-pressions of Tspan8 and ITGA5(P＜0.05).TNM stage Ⅲ-Ⅳ(HR=2.289,95％CI:1.519-3.447),other molecular subtypes(HR=2.622,95％CI:1.744-3.942),Tspan8 positive expression(HR=3.622,95％CI:2.159-6.077)and ITGA5 positive expression(HR=3.142,95％CI:2.022-4.884)were risk factors for ax-illary lymph node metastasis in breast cancer patients(P＜0.05).Conclusion Tspan8 and ITGA5 are highly expressed in breast cancer patients,which are related to the clinicopathological characteristics and prognosis of patients.

Integrin α5 silencing inhibits proliferation, invasion and metastasis of human liver cancer Bel-7404 cells / 南方医科大学学报

OBJECTIVE@#To analyze the association of integrinα5 (ITGA5) with grading of liver cancer and the overall patient survival and investigate the effects of integrin α5 (ITGA5) silencing on the proliferation, invasion and migration abilities of human liver cancer Bel-7404 cells.@*METHODS@#UALCAN was used to analyze the expression of ITGA5 in liver cancer tissues and normal tissues, and expression in different grades of liver cancer tissues. GEPIA was used to analyze the relationship between ITGA5 expression and the survival of liver cancer patients through.The ITGA5 shRNA lentiviral vector was used to infect Bel-7404 cells to establish a cell line with stable ITGA5 silencing verified by Western blotting. Plate clone formation assay and Transwell assay were used to detect the proliferation, invasion and migration of Bel-7404 cells. The correlation between ITGA5 and PI3K in liver cancer tissues and control tissues was analyzed using Oncomine cancer specimen database.@*RESULTS@#The expression of ITGA5 was significantly higher in liver cancer than in normal tissues ( < 0.05). The expression of ITGA5 was significantly lower in grade 1 than in grade 2 liver cancer, and also lower in grade 2 than in grade 3 liver cancer ( < 0.05). The patients with high ITGA5 expression had a significantly lower overall survival rate than those with low ITGA5 expression ( < 0.05). Plate clone formation assay showed that the clone formation rate was significantly lowered in Bel-7404 cells with ITGA5 silencing compared with the blank and negative control cells ( < 0.05). ITGA5 silencing significantly attenuated the migration of Bel-7404 cells as shown by Transwell assay ( < 0.05). ITGA5 and PI3K were both highly expressed and showed a positive correlation in liver cancer tissues ( < 0.05).@*CONCLUSIONS@#ITGA5 is closely related to the progression of liver cancer and the patients' prognosis. ITGA5 silencing inhibits the proliferation, invasion and migration of liver cancer cells. ITGA5 promotes the liver cancer growth and metastasis possibly by regulating the PI3K signaling pathway.

The effects of mitogen-activated protein kinase on EGF-induced expression of integrin α_5 in human RPE cells / 眼科研究

Background Human retinal pigment epithelial(RPE) cells play a critical role in the pathogenesis of proliferative vitreoretinopathy(PVR) and other related ocular diseases.Research demonstrated that epidermal growth factor(EGF) stimulates activation of RPE cells and therefore causes PVR,and integrin α_5 mediates the adhesion of cells and EGF.Objective This study aims to investigate the role of mitogen-activated protein kinase(MAPK) in regulation of EGF on integrin α_5 expression in human RPE cells.MethodsHuman RPE cells strain(CRL2302) was cultured in DMEM/F12 with 10% calf serum and passaged in 0.25% trypsin.Cultured cells were divided into DMEM/F12control group,20μmol/L PD98059 +10 ng/mL EGF＋DMEM/F12(PD98059) group and 10 ng/mL EGF＋DMEM/F12(EGF) group.The expression of integrin α_5 protein in CRL2302 cells was detected by RT-PCR and calculated as α_5 mRNA/β-actin mRNA,and the expression of integrin α_5 mRNA in CRL2302 cells was detected evaluated by flow cytometry.The MAPK phosphorylation level in each group of human RPE cells was tested by Western blot.ResultsThe expression value of integrin α_5 mRNA was 0.93±0.06 in the control group,1.06±0.07 in PD98059 group and 1.97±0.09 in EGF group,showing a significant difference among the three groups(F=458.896,P<0.01).The fluorescence intensity of integrin α5 protein in CRL2302 cells after 24 hours was 1.94±0.22,4.56±0.25,2.39± 0.14 in three groups respectively with a significant difference(F=21.05,P<0.05).After 30 minutes of culture,Western blot result showed that the strongest phosphorylation levels of ERK1/2 activation in EGF group and the weakest phosphorylation levels of ERK1/2 activation in the control group;While that in PD98059 group was significantly stronger than control group and weaker than EGF group(F=143.14,P<0.01).ConclusionEGF stimulates activation of ERK1/2 pathway in human RPE cells and the expressions of integrin α_5 mRNA and protein in human RPE cells in vitro.