Expression And Significance Of TROP2 In Lung Adenocarcinoma

The first chapter: TROP2 expression in lung adenocarcinoma tissue and its relationship with clinical pathological featuresObjective: To study the TROP2 of the expression and clinical significance in human lung adenocarcinoma cells, human lung adenocarcinoma tissue.Methods: TROP2 expression of 68 cases of lung adenocarcinoma tissues and corresponding 20 tissue adjacent to carcinoma were detected by immunohistoch-emical, to tatistical analysis of its relationship with clinical pathological featur es; RT-q PCR test specimen:38 cases of lung adenocarcinoma and tissue adjacen-t to carcinoma and lung adenocarcinoma cancer cell H1299,SPCA-1,PC-9,A549,bronchial epithelial cell line HBE TROP2 m RNA expression,Western Blot examination TROP2 protein expression in lung adenocarcinoma cancer cells.Results: 1. TROP2 in tissue adjacent to carcinoma did not or unusual expression, the expression of the main parts in the bronchial mucosa, alveolar wall, the positive expression rate was 15%(3/20). TROP2 in lung adenocarcinoma tissues expressed in the cytoplasm and membrane of tumor cells, for the size of sandy particles and the nucleus is not coloring; TROP2 positive expressionrate of 54.4%(38/68) higher than the tissue adjacent to carcinoma(P=0.0002). Lung adenocarcinoma group TROP2 positive expression rate is higher thantissue adjacent to carcinoma. Lung adenocarcinoma group TROP2 expression and tumor differentiation degree(P=0.013),lymph node metastasis(P=0.038) and the TNM staging(P=0.012) were positively correlated, but with age, sex, tumor size,Tumour Stage has nothing to do(P=0.584,0.112, 0.685, 0.414);2. Expression of m RNA TROP2 in the adjacent tissues(0.159± 0.009), while the relative expression of m RNA TROP2 in lung adenocarcinoma was(1.153±0.077),which was significantly higher than that in the adjacent tissues(P<0.005).3. RT-q PCR to detect lung adenocarcinoma cells of TROP2 m RNA expression,the results showed that the expression of TROP2 quantity order from high to low is: PC-9>SPCA-1>H1299>A549>HBE. Western Blot detection TROP2 pro-tein expression, the results showed: the expression of TROP2 amount from high to low in turn for PC-9>SPCA-1>H1299>A549>HBE.Conclusion: 1. TROP2 in lung adenocarcinoma tissue increased, there were significantly different compared with the expression of tissue adjacent to carcinoma(P=0.0002); 2.The expression of TROP2 and tumor differentiation degree(P =0.013),lymph node metastasis(P=0.038) and the TNM staging(P=0.012) were positively correlated, but with age, sex, tumor size, Tumour Stage has nothing to do; 3. TROP2 high expression in lung adenocarcinoma cancer cells, and higher in the PC-9 cells.The second chapter: TROP2 gene expression plasmid and TROP2-sh RNA plasmids Build and screening of lung adenocarcinoma cancer cellObjective: To research on the mechanisms of bortezomib in lung adenocarcinoma,we build TROP2 gene expression plasmid pc DNA3.1-TROP2 and interference plasmid sh RNA-TROP2 plasmid, and build TROP2 high/low expression of lung adenocarcinoma cancer cell model.Methods: 1. Through gene cloning people TROP2 gene cDNA sequence, the cloning,enzyme digestion, connecting carrier pc DNA3.1, build TROP2 gene expression plasmid pc DNA3.1-TROP2, and build express TROP2 lung adenocarcinoma cell line A549 steady turn, using RT-q PCR and Western Blot method appraisal expressproduct;2. For people TROP2 gene PLKO.1 carrier construction three of TROP2-sh RNA plasmids, express product identified by Western Blot, transfectionliposome transfection technique used in PC-9 cells and RT-q PCR and Western Blot detecting expression product, select sh RNA interference effect is best. And Build the lower expression of TROP2 lung adenocarcinoma cancer cell model, using Western Blot method to identify expression product.Results: 1. The successful build pcDNA3.1-TROP2 expression plasmid; 2. On the basis of A549 cells successfully constructed stable expression of TROP2 lung adenocarcinoma cancer cell system model; 3. The successful construction of plasmid TROP2 TROP2 interference-sh RNA, by Western Blot and RT-q PCR appraisal show TROP2-sh RNA-2 the interference effect of the best; 4. Will TROP2-sh RNA-2transfection into PC-9 lung adenocarcinoma cancer cell, successful build the lower expression of TROP2 lung adenocarcinoma cancer cell model.Conclusion: Successful construction of plasmid pcDNA3.1-TROP2 expressionplasmid and TROP2-sh RNA interference, and the transient transfection afteridentified the best sh RNA interference effect. And build TROP2 gene expression and the lower expression of lung adenocarcinoma cancer cell model.The third chapter: TROP2 effects on lung adenocarcinoma cancer cell biological featureObjective: To study the TROP2 effects on lung adenocarcinoma cancer cell biological feature.Methods: TROP2 expression plasmid pcDNA3.1-TROP2, construct the overexp-ression of TROP2 lung adenocarcinoma A549 cell model; Using plasmid TRO-P2-sh RNA TROP2 interference, low expression of TROP2 lung adenocarcinoma cancer cell model was constructed. CCK-8 method is used to test, scratch test, cell invasion experiment, cell apoptosis, in vitro observation TROP2 expr-ession after the change, the lung adenocarcinoma cancer cell proliferation, mig-ration, invasion, and apoptosis biological feature.Results: Confirm overexpression of TROP2 CCK-8 method to promote A549 cell proliferation, inhibition of TROP2 expression after the inhibition of proliferation of lung gland cancer cells PC-9; Scratch the experiment demonstrated that expression TROP2 could accelerate the A549 cell migration ability, inhibit TROP2 expression to their PC-9 cells after migration ability; Cell invasion experiment(Transwell Chambers) suggest that TROP2 after expression canpromote effectively the invasive ability of A549 cells, interfere with TROP2 expression can effectively restrain the PC-9 cells after attacking ability; Apoptosis results: after TROP2 expression can effectively inhibit the apoptosis of A549 cells, interference can effectively promote the PC-9 cell after TROP2 expression apoptosis;Conclusions: TROP2 promote lung adenocarcinoma cancer cell proliferation,migration and invasion, and effectively suppress the lung adenocarcinoma cancer cell apoptosis.