| Objective : To explore the relationship and mechanism between epithelial shear regulated protein 1(ESRP1)expression and epithelial mesenchymal transformation(EMT)in NSCLS.Methods:1.A total of 65 paraffin-embedded surgical specimens and fresh tissue specimens of patients with non-small cell lung cancer diagnosed by pathology in宁夏医科大学general hospital from 2015 to 2019 were collected as the lung cancer group.A total of 25 cases of paraffin-embedded surgical specimens and fresh tissue specimens from lung tissues above 5cm at the edge of tumor lesions were selected as the para-carcinoma group.A total of 20 paraffin embedded specimens and fresh lung tissue specimens from patients with carcinoma in situ and atypical hyperplasia were selected as the precancerous group.A total of 17cases of paraffin-embedded specimens and fresh hydrothorax sediment samples with metastatic lung cancer were included in the lung metastatic cancer group.At the same time,30 cases of paraffin embedded surgical specimens and fresh tissue specimens with pathologically confirmed benign lung lesions who were hospitalized for lobectomy or segectomy at the same time were selected as the control group.The expression of ESRP1,related transcription factor Snail,Twist and EMT-related markers(E-ca,N-ca,Vimentin,a-SMA)in lung tissue samples ofthe control group,precancerous lesion group,lung cancer group,paracancer group and lung metastatic cancer group were detected by immunohistochemistry and western blots,respectively.To analyze the correlation between the expression of ESRP1 protein and the clinical parameters of NSCLC:age,sex,smoking history,tumor stage,differentiation degree and lymph node metastasis,and the relationship between the expression of ESRP1 protein and the prognosis and survival of patients.2.Human lung adenocarcinoma cell line(A549)was cultured in vitro.Transforming growth factor TGF β-1 was used to treat cells at different concentrations and at different time points.The expressions of ESRP1,related transcription factor Snail,Twist and EMT-related markers(E-ca,N-ca)were detected by western blots.Results1.Study on the expression of ESRP1,EMT-related transcription factors and EMT-related marker proteins in the precancerous lesions,lung cancer,paracancer and metastatic lung cancer groups of non-small cell lung cancer1.1Comparative study on expression of emt-related marker proteins(E-ca,N-ca,Vimentin,a-SMA)in non-small cell lung cancer precancerous lesions,lung cancer,paracancer and metastatic lung cancer:Expression of EMT-related marker proteins in non-small cell lung cancer group,precancerous lesion group and control group:(1)Immunohistochemical staining and Western Blotting showed that the EMT intermediate phenotype markers in the precancerous lesion group and the non-small cell lung cancer group were higher than those in the control group.The expression of E-ca,a marker of epithelial phenotype,was lower than that of the control group,and the differences were statistically significant(P< 0.05).However,there was no significant difference in EMT expression of a-SMA and N-cabetween the precancerous lesion group and the non-small cell lung cancer group(P>0.05).(2)Expression of EMT-related proteins in non-small cell lung cancer group,paracancer group and metastatic lung cancer group:Immunohistochemical staining and western blotting results showed that the expression levels of the EMT intermediate phenotype markers a-SMA and N-ca in the lung cancer group and the lung metastatic cancer group were higher than that in the para-cancer group,and the differences were statistically significant(P<0.05).There was no significant difference in EMT expression between the lung cancer group and the lung metastatic cancer group(P>0.05).1.2.Comparative study on the expression of ESRP1 and related transcription factor Snail and Twist in the groups of non-small cell lung cancer:(1)Expression of ESRP1 and related transcription factor Snail and Twist in non-small cell lung cancer group,precancerous lesion group and control group:Immunohistochemical staining and western blotting results showed that the expressions of ESRP1 and emt-related transcription factor Snail and Twist in the precancerous lesion group and the non-small cell lung cancer group were higher than those in the control group,and the differences were statistically significant(P<0.05).(2)Expression of ESRP1 and related transcription factor Snail and Twist in non-small cell lung cancer group,paracancer group and lung metastatic cancer group:Immunohistochemical staining and western blotting results showed that ESRP1 expression in the lung cancer group was higher than that in the paracancer group and the metastatic cancer group,and the differences were statistically significant(P<0.05).There was no significant difference in ESRP1 expression between paracancer group and lung metastatic cancer group(P>0.05).The expressions of Snail and Twist in the lung cancer group and the lung metastatic cancer group were all higher than those in the para-cancer group,and thedifference was statistically significant(P<0.05),while the expressions of Snail and Twist in the lung cancer group and the lung metastatic cancer group were not statistically significant(P>0.05).1.3.Analysis of the expression of ESRP1 in lung tissues of NSCLC group and the clinical parameters of patients with lung cancer: in NSCLC tissues,the expression of ESRP1 was closely related to smoking history(P<0.05),tumor differentiation degree(P<0.05),tumor stage(P<0.05),and distant metastasis(P<0.05).Moreover,kaplan-meier curve showed that the overall survival rate of patients with high ESRP1 expression was significantly better than that of patients with low or negative ESRP1 expression(P<0.05).COX survival analysis showed that ESRP1 was an independent risk factor affecting the prognosis of patients with non-small cell lung cancer.2.Expression of EMT and EMT-related transcription factors in human lung adenocarcinoma cell A549 by TGF-β12.1Expression of EMT-related markers in A549 cells induced by TGF-β1 :TGF-β 1 stimulated A549 cells at different concentrations and time points,western blot detected EMT-related marker proteins: the expression level of N-ca protein was higher than control group(P<0.05),while the expression level of E-ca protein was lower than control group(P<0.05).2.2Expression of ESRP1,Snail and Twist in A549 cells treated by TGF-β1:TGF-β 1 stimulated A549 cells at different concentrations and time points : In western blot detection,ESRP1 expression was lower than that of control group(P<0.05),while Snail and Twist expression was higher than that of control group(P<0.05).Conclusion1.The occurrence and development of non-small cell lung cancer all havedifferent degrees of epithelial mesenchymal phenotypic transformation(EMT):The transformation of the E phenotype to the M phenotype was observed in the precancerous lesions,the cancerous tissues and the metastatic lung cancer.2.In the development and development of NSCLC,ESRP1 and related transcription factor Snail and Twist are expressed in different degrees.In other words,ESRP1 and related transcription factor Snail and Twist were highly expressed in precancerous lesions of lung cancer,cancer tissues,and ESRP1,and the expression of ESRP1 decreased in metastatic lung cancer and paracancer.The related transcription factor Snail and Twist were up-regulated in metastatic lung cancer.Moreover,the expression of ESRP1 is closely related to the degree of tumor differentiation and TNM stage,suggesting that ESRP1 and related transcription factor Snail and Twist may be involved in the occurrence and development of lung cancer through some signaling pathway.3.The positive expression of ESRP1 is closely related to the prognosis of NSCLC patients and may be a good prognostic marker.4.TGF-β1 up-regulated the expression of EMT transcription factor Snail and Twist and inhibited the expression of EMT regulatory factor ESRP1,thus inducing the emergence of EMT in human lung adenocarcinoma cells.This suggests that TGF-β 1-Snail/ Twist-ESRP1-EMT signaling pathway plays an important role in the development of EMT in human lung adenocarcinoma cells. |
PDF Full Text Request