| Objective:To detect the expression of Notch1,NICD1 and Hey1 in various lung cancers and adjacent normal lung tissues,and to compare the differential expression of various indicators in different types of lung cancer,and to analyze:(1)The correlation between each index and clinicopathological features and prognosis of various types of lung cancer patients;(2)The relationship between Notch1 expression and Notch1 activation,Notch1 activation and Hey1 expression in various types lung cancer;(3)The correlation between the relative expression of Hey1 mRNA and protein expression in non-small cell lung cancer.It provides a theoretical basis for the early diagnosis,prognosis and treatment of lung cancer.Methods:1.Tissue microarray was used to detect the expression of Notch1,NICD1 and Hey1 in 118 cases of lung squamous cell carcinoma,128 cases of lung adenocarcinoma,69 cases of small cell lung cancer and 117 cases of adjacent normal lung tissue paraffin specimens by immunohistochemical En Vision method.The results were interpreted by immunoreactive integral method and H-score scoring method.The expressions of Notch1,NICD1 and Hey1 in various lung cancers and adjacent normal lung tissues were compared,and the relationship between each index and the clinicopathological features and prognosis of various lung cancer patients was analyzed.2.Real-time quantitative PCR(qRT-PCR)and immunohistochemical En Vision method were used to detect the relative expression of Hey1 mRNA and protein expression in 34 cases of non-small cell lung cancer(28 cases of lung adenocarcinoma,6 cases of lung squamous cell carcinoma)and 21 cases of adjacent normal lung tissues,and the correlation between Hey1 protein expression and relative expression of mRNA was analyzed.Results:1.The interpretation of the results of immunohistochemistry(IHC)in this experiment was performed using the immune response integration method and the H-score scoring method.In lung squamous cell carcinoma,both methods showed that the positive expression of Notch1,NICD1 and Hey1 was significantly higher than that of adjacent normal lung tissues(P<0.05).In lung adenocarcinoma tissues,the immune response integration method showed that the positive expression rate of Notch1 was not significantly different from that of normal lung tissues(P>0.05);H-score score showed that the positive expression of Notch1 was significantly lower than that of adjacent normal lung tissues(P<0.05);both methods showed that the positive expression of NICD1 and Hey1 was not significantly different from that of adjacent normal lung tissues(P>0.05).In small cell lung cancer,both methods showed that the positive expression of Notch1 was not significantly different from that of adjacent normal lung tissues(P>0.05);The positive expression of NICD1 and Hey1 was significantly higher than that of adjacent normal lung tissues(P<0.05).2.The immunohistochemical results of different interpretation methods were analyzed by the corresponding test methods.The results of the immune response integration method were analyzed by chi-square test,and the results of H-score scoring were analyzed by non-parametric Mann-Whitney U test.Chi-square test showed that the expression of Notch1 in lung squamous cell carcinoma was positively correlated with TNM stage and lymph node metastasis.The positive expression of NICD1 was not associated with clinicopathological features.The positive expression of Hey1 was positively correlated with TNM stage and negatively correlated with tumor differentiation.The positive expression of Notch1,NICD1 and Hey1 in lung adenocarcinoma patients was not associated with any clinicopathological features.The non-parametric Mann-Whitney U test showed that the positive expression of Notch1 in lung squamous cell carcinoma was positively correlated with TNM stage and lymph node metastasis.The positive expression of NICD1 was not associated with clinicopathological features.The positive expression of Hey1 was positive with TNM stage and lymph node metastasis.The positive expression of Notch1,NICD1 and Hey1 in lung adenocarcinoma patients was not associated with any clinicopathological features.Both tests showed that the expression of Notch1,NICD1 and Hey1 in small cell lung cancer patients was not associated with any clinicopathological features.3.Spearman correlation analysis,lung squamous cell carcinoma,both methods of interpretation showed that Notch1,NICD1 and Hey1 were positively correlated;while in lung adenocarcinoma tissues,the immune response integration method showed that Hey1 was positively correlated with Notch1 and NICD1,Notch1 not related to NICD1,Notch1 was highly expressed compared with NICD1;H-score scoring showed that Notch1,NICD1 and Hey1 were positively correlated.In small cell lung cancer tissues,the immune response integration method showed that NICD1 was positively correlated with Notch1 and Hey1,Notch1 was not associated with Hey1;H-score scoring method showed that Notch1 was positively correlated with Hey1,and NICD1 was not correlated with Notch1 and Hey1,and NICD1 was highly expressed compared to Notch1..4.Both interpretation methods and survival analysis showed that the expressions of Notch1,NICD1 and Hey1 in lung squamous cell carcinoma,lung adenocarcinoma and small cell lung cancer were not associated with overall survival.5.The relative expression of Hey1 mRNA in lung adenocarcinoma patients was significantly lower than that in adjacent normal lung tissues,which was not related to clinicopathological features.The relative expression of Hey1 mRNA in lung squamous cell carcinoma patients was not significantly different from that in adjacent normal lung tissues(P>0.05),Because the number of cases of lung squamous cell carcinoma is too small,the relationship between the relative expression of Hey1 mRNA and the clinicopathological features of patients has no statistical value.6.The relative expression of Hey1 mRNA in lung squamous cell carcinoma and lung adenocarcinoma was inconsistent with protein expression.Conclusion:1.Immunological integration and H-score scoring were used to interpret the results of immunohistochemistry,and different results may be obtained.But the overall trend is the same.The H-score scoring method has a small subjective error and is more scientific and rigorous.2.The expression of Notch1,NICD1 and Hey1 is different in different types of lung cancer.Hey1 is an important factor in the occurrence of lung squamous cell carcinoma and small cell lung cancer,which may promote the occurrence of lung squamous cell carcinoma and small cell lung cancer.3.Notch1 and Hey1 can be used as auxiliary indicators to determine the pathological grade and malignancy of lung squamous cell carcinoma,and guide the choice of clinical treatment options.4.The activation mechanism of Notch1 may be different in different types of lung cancer.The expression of Hey1 in lung squamous cell carcinoma and lung adenocarcinoma may be induced by activation of Notch1,whereas in small cell lung cancer,Hey1 expression may be activated by other signaling pathways.5.The expression of Hey1 protein in non-small cell lung cancer may be regulated by mRNA level and translation level,resulting in the inconsistent expression of Hey1 mRNA and protein expression. |
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