| Research objectives:1.To detect the expression of RAGE,S100 P,miR-21 and STAT3/Pim1 pathway in non-small cell lung cancer tissues and adjacent normal lung tissues.2.To analyze the relationship between RAGE,S100 P and the clinicopathological characteristics of non-small cell lung cancer patients;to discuss the role of RAGE,S100 P play in the non-small cell lung cancer.3.To analyze the correlation between RAGE,S100 P and miR-21,STAT3/Pim-1 signaling pathway.To investigate the possible mechanisms of the involvement and interactions between mentioned-above factors in non-small cell lung cancer.Research materials and methods:1.Clinical tissue specimens: 40 patients with non-small cell lung cancer were from the affiliated hospital of Qingdao university.Fresh tissue samples were obtained from each patient after thoracic surgery,including cancer tissues and paired non-tumor tissues.2.The expression of RAGE,S100 P in 10 cases of cancer tissues and adjacent normal lung tissues were detected by immunohistochemical technique,their location and expression in cells were also analyzed.3.RAGE,S100 P,STAT3 and Pim1 protein in 12 cases of cancer tissues and paired non-tumor tissues were then measured by Western blot.4.Corrspondingly,in 40 cases,the mRNA expression levels of RAGE,S100 P,miR-21,STAT3 and Pim1 in cancer tissues and paired normal tissues were measured by Real-time quantitative PCR(RT-qPCR).5.Statistical analyses using Graphpad Prism 6: T-test was used to analyze the differentces of RAGE,S100 P,miR-21,STAT3 and Pim1 mRNA,as well as to analyze the differences of the expression of RAGE,S100 P between different clinicophalogical parameters of patients,including age,gender,smoking status,degree of differentiation,lymph node metastasis and TNM staging of non-smallcell lung cancer.;Pearson correlations was used to analyze the correlations between RAGE,S100 P and miR-21,STAT3/Pim1 pathway.P < 0.05 was considered to be of statistical significance.Research results:1.Immunochemical detected the the expression of RAGE,S100 P in 10 paired non-small cell lung cancer tissues and normal tissues.RAGE,S100 P displayed expression location in cancer tissues normal tissues,more specifically,a majority of RAGE was situated in the cytoplasm with a few in the cell membrane,a majority of S100 P was situated in the cytoplasm with a few in the nucleus and cell membrane.2.Western blot detected the expression of RAGE,S100 P,STAT3 and Pim1 protein in 12 paired cancer tissues and normal tissues.Compared with the ajacent normal tissues,the expression of RAGE was significantly decreased in cancer tissues;however,the expression of S100 P,STAT3 and Pim1 was significantly increased in cacer tissues.3.RT-qPCR detected the expression of RAGE,S100 P,miR-21,STAT3 and Pim1 mRNA in 40 paired cancer tissues and normal tissues.Compared with the ajacent normal tissues,the expression of RAGEmRNA was significantly decreased in cancer tissues;however,the expression of S100 P,miR-21,STAT3 and Pim1 mRNA was significantly increased in cacer tissues.4.The expression of RAGE,S100 P is not associated with age,gender,and family history of lung cancer patients.Both of them were associated with smoking status,the level of RAGE in patients with smoking history was significantly lower than that in patients with no smoking history,the level of S100 P in patients with smoking history was significantly higher than that in patients with no smoking history.The expression of RAGE and S100 P were related to the anatomical classification,histopathological type,the degree of differentiation of lung cancer,lymph node metastasis.The level of RAGE in central lung cancer tissues is significantly lower than that in peripheral lung cancer tissues,and lower in squamous tissues than that in adenocarcinoma cell carcinoma tissues;the level of RAGE in poorly differentiated cancer tissues was significantly lower than that in high-grade differentiated cancer tissues;its level in cancer tissues with lymph node metastasis was significantly lower than that in cancer tissues without metastasis;moreover,the level of RAGE in cancer tissues of TNM stage II was lower than that in TNM stage I cancer tissues.The level of S100 P in central lung cancer tissues is significantly higher than that in peripheral lung cancer tissues,and higher in adenocarcinoma tissues than that in squamous cell carcinoma tissues;the level of S100 P in cancer tissues with higher differentiation was signinificantly higher in that in less differentiated cancer tissues,and higher in cancer tissues with lymph node metastasis than that in cancer tissues without lymph node metastasis;5.In non-small cell lung cancer tissues,the expression level of S100 P was negatively correlated with RAGE;the expression level of STAT3 was negatively correlated with RAGE;the expression level of miR-21 was negatively correlated with RAGE;the expression level of STAT3 was positively correlated with S100P;the expression level of Pim1 was positively correlated with S100P;the expression level of miR-21 was positively correlated with S100P;the expression level of Pim1 was positively correlated with STAT3;the expression level of miR-21 was positively correlated with Pim1.Conclusions:1.In non-small cell lung cancer tissues,the expression level of RAGE was down-regulated but the expression level of S100 P,miR-21 and STAT3/Pim1 signaling pathways was significantly up-regulated.2.The expression of RAGE,S100 P was related to the clinical and pathological features of patients with non-small cell lung cancer,both of which may play an important role in the occurrence,invasion and metastasis of lung cancer.3.The expression of RAGE,S100 P was related to the expression of miR-21 and STAT3/Pim1 signaling pathways,suggesting that the functions of RAGE and S100 P may be controled or mediated through miR-21 regulated by STAT3/Pim1 pathway,all of them participate in the development of lung cancer. |
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