Mechanism Of Interaction Between TM4SF1 And Jak2-Stat3 Signaling Pathway In Lung Cancer Cells And Expression Analysis In Non-small Cell Lung Cancer

Objective To explore the expression of transmembrane 4 superfamily member 1(TM4SF1)in non-small cell lung cancer(NSCLC)and try to clarify its inherent relationship with clinicopathological characteristics of NSCLC patients contact.Build a lentiviral expression vector that overexpresses TM4SF1,use it to transfect lung cancer A549 cells,and explore the role of its migration function with lung cancer cell A549 and the effect of JAK2-STAT3 signaling pathway on downstream Sox2 expression level in lung cancer A549 cell line.Methods A total of 61 cases of cancer tissues,corresponding tissues adjacent to cancer,and normal lung tissues removed due to benign lung disease were collected from 61 patients with NSCLC(June 2018-December 2018).Extract the mRNA in cancer tissue,adjacent tissue and benign lung tissue respectively,and then use real-time immunofluorescence quantitative PCR(qRT-PCR)method to determine the expression of TM4SF1 at the mRNA level after the synthesis of c DNA,and explore the expression of RNA in TM4SF1 in different fresh specimens Differences and the relationship between different expression levels and clinicopathology.Western blot was used to determine the expression of TM4SF1 at the protein level in cancer tissues,adjacent tissues,and benign lung tissues,and to explore the significance of the difference in expression.The TM4SF1 gene was overexpressed in lung cancer A549 cells using lentiviral transfection.Real-time fluorescence quantitative PCR(qRT-PCR)and Western Blot experiments were used to determine the successful expression of the target gene TM4SF1.The high expression of TM4SF1 caused changes in the migration ability of A549 by the Transwell method.After adding JAK2-STAT3 signal pathway interference reagent AG490,real-time immunofluorescence quantitative PCR(qRT-PCR)method was used to analyze the transfection group,blank group,negative control group and their corresponding treatment groups TM4SF1,JAK2-STAT3 signal pathway Stat3 and downstream Sox2 The RNA expression level of the gene.At the same time analyze the correlation of these three indicators.Results The study on the expression of TM4SF1 in fresh specimens showed that the mRNA level of TM4SF1 in cancer tissues was higher than that of the corresponding adjacent tissues.The expression level of TM4SF1 in cancer tissues with mRNA levels was 4.90 times that of adjacent tissues.The difference in expression levels was statistically significant Academic significance(t =5.752,P=(0.000<0.05);and significantly higher than normal lung tissue specimens,5.17 times that of normal lung tissue(t =3.541,P = 0.001 < 0.05),while mRNA-level adjacent tissues There was no significant difference in the expression of TM4SF1 from benign lung tissue(P>0.05).The expression of TM4SF1 at the mRNA level is closely related to the degree of differentiation,tumor size,positive rate of lymph nodes,and clinical stage(P <0.05).The high expression of TM4SF1 mRNA may show a higher tendency and positive rate of lymph node positive Late clinical staging.However,the expression of TM4SF1 at the mRNA level has not shown a significant correlation with the patient’s age and gender(P> 0.05).TM4SF1 was successfully overexpressed in lung cancer cell A549.Real-time fluorescence quantitative PCR(qRT-PCR)and Western-blot method were used to detect the expression level of TM4SF1 in the transfection group.The mRNA expression of TM4SF1,Sox2 and Stat3 in the transfection group was significantly higher than that in the negative control group(P <0.01)and the blank group(P <0.01),and the expression level of TM4SF1 protein in the transfection group was significantly higher than that of the negative control group and the blank group(P = 0.001 <0.01).After adding JAK2-STAT3 signaling pathway interference reagent AG490 to the medium,the three groups of cells were continuously cultured The expression levels of Stat3 and downstream Sox2 in the drug interference group were significantly lower than those in the untreated group(P <0.05),indicating that JAK2-STAT3 may be a signaling pathway for TM4SF1 to regulate its downstream Sox2 expression level.After overexpressing TM4SF1,Transwell was used to verify the enhanced movement of lung cancer A549 cells,and the results showed that its migration level was significantly improved.The expression of TM4SF1,Stat3 and Sox2 at the mRNA level showed a positive correlation trend(P <0.01).Conclusion The expression of TM4SF1 in NSCLC is high,and the larger the mass,the lower the degree of differentiation.The corresponding positive rate of lymph nodes and the later clinical stage.The expression of TM4SF1 at the mRNA level is high,which makes us malignant to NSCLC.There is a new standard for grading.After overexpression of this gene,it can significantly increase the migration level of lung cancer A549 cells,and may affect or even upregulate the expression of Sox2 below through the JAK2-STAT3 pathway.The expression of TM4SF1,Stat3 and Sox2 was positively correlated in A549 cells.This pathway may be a key pathway for TM4SF1 to promote the occurrence and development of NSCLC and distant metastasis,indicating that TM4SF1 may play an important role in the occurrence and development of NSCLC and hopefully serve as a new target for cancer treatment.