| With the increase of smoking and the deterioration of the living environment in modern society, incidence of lung cancer increased significantly. It has occupied the first overall cancer incidence, threaten human health and life and become one of the largest cancer. In recent decades increasingly younger age of on set, and male lung cancer incidence and mortality of malignant tumor, female morbidity, mortality ranks second after breast cancer, a serious threat to human health.Purpose: Pulmonary sarcomatoid carcinoma and squamous cell carcinoma of transforming growth factor-β1, the expression of EMT related proteins analyzed their relevance. And to explore the relationship between lung sarcomatoid carcinoma of TGF-β1 and vasculogenic mimicry for clinical survival, influence and prognosis. EMT phenotypic changes and affect cell movement ability of lung squamous cells, and describes EMT increase in lung cancer cell invasion and migration by in vitro TGF-β1-induced, VM formation mechanism.Methods: 1)Analysis of the Tianjin Medical University Cancer Hospital from January 1995 to December 2010 before surgery did not receive chemotherapy treatment of pulmonary sarcomatoid carcinoma in 41 patients, lung squamous cell carcinoma 79 cases of clinical data, summarize the follow-up results. Respectively, lung cancer and pulmonary sarcomatoid squamous cell carcinoma were TGF-β1 and EMT related indicators E-cadherin, Vimentin, Twist1 immunohistochemical staining to analyze the relationship between the two groups in the TGF-β1 and EMT related proteins. And human lung tissue sections sarcomatoid cancer cases were 41 cases of CD31 and PAS double staining analysis prognosis of TGF-β1 and VM between sarcomatoid carcinoma of the lung. Finally, VE-cadherin, MMP-2 protein immunohistochemical analysis of the mechanisms of pulmonary sarcomatoid carcinoma in VM formation. 2)By culturing SK-MES-1 lung cancer cell lines, and the use of TGF-β1 induced squamous cell carcinoma of the lung. Observe whether EMT occurs, while promoting the formation of VM. Some methods use cell function studies were observed after lung cancer cell invasion ability of induced changes in migration speed changes, as well as 3D Pipes forming ability. 3)Apply Western Blot, RT-PCR, immunofluorescence experiments from the protein and m RNA levels detected E-cadherin and Vimentin expression. Explore EMT occurs after induction of lung squamous cell carcinoma from the mechanism, thus promoting the formation of VM. 4) Western Blot detect differential expressions of key proteins to form VM VE-cadherin induced by TGF-β1. 5)Gelatin zymography analysis through TGF-β1 inducing MMP-2 and MMP-9 activity of matrix metalloproteinases.Results:1)the expression of EMT associated protein and TGF-β1 in pulmonary sarcomatoid carcinoma and lung squamous cell carcinomaBased on 41 cases of pulmonary sarcomatoid carcinoma, 79 cases of squamous cell carcinoma EMT-associated protein immunohistochemical staining, found that compared with lung squamous cell carcinoma, lung sarcomatoid carcinoma decreased expression of E-cadherin, Vimentin expression was significantly enhanced between the two groups Compare the difference was statistically significant. Followed by TGF-β1, Twist1 immunohistochemical staining found in the lung sarcomatoid carcinoma and squamous cell carcinoma of the lung in both groups, TGF-β1 and the difference was statistically significant Twist1 of.2) Relationship between TGF-β1 and VM formation41 cases of lung sarcomatoid cancer patient of TGF-β1 expression was 34.1%(14/41). VM positive expression rate of TGF-β1 group 70%(7/10); VM positive expression of TGF-β1-negative group was 22.6%(7/31). TGF-β1 has a statistically significant difference between the VM-positive group and the positive expression rate of VM-negative group. Within the TGF-β1 protein was located in the cytoplasm of lung sarcomatoid carcinoma, emerge a brown granules. Survival analysis showed that, TGF-β1 positive group overall survival time shorter than the negative group.3)TGF-β1-induced lung squamous cell carcinoma of the three-dimensional culture in vitro confirmed that it promoted the VM formation of lung squamous cell angiogenesis, enhanced invasion and migration of lung squamous carcinoma cells.After TGF-β1-induced l lung squamous carcinoma cells after the Matrigel able to form a three-dimensional culture models VM, and the pipeline structure is more complicated. Transwell chamber in vitro invasion assay found that different concentrations of TGF-β1-induced lung squamous carcinoma cells at different times, compared with the control group, high concentration and inducing a long time, the number of cells in the experimental group are the more penetrating, statistics showed a statistically significant difference. Migration experiment found that, compared with the control group, after TGF-β1-induced lung squamous carcinoma cells at the same time, the number of migrating cells are more, statistically significant difference. Scratch experiment also found that, TGF-β1-induced 10 d, 15 d lung squamous cell groups in the 48 h wound healed completely. The wound healing rate were higher than control group, the difference was statistically significant. Further evidence of lung squamous cell induction after its invasion and mobility increased significantly compared with the control group.4)In vitro TGF-β1-induced lung squamous cells EMT, promote the formation of VM Immunofluorescence experiments confirmed that SK-MES-1 cells not only in morphology occurred changes after TGF-β1-induced, but also occurred in EMT, there have been changes in antigenic phenotype, the transition from epithelial to mesenchymal phenotype type. RT-PCR and Western Blot detect protein and m RNA expression after induction confirmed squamous cell lung was significantly E-cadherin downregulated and Vimentin upregulated in protein and m RNA levels after induction. These results further confirm the occurrence of lung squamous cell plasticity EMT-like change, thus contributing to the formation of vascular mimicry.5)A key protein of VM, VE-cadherin and matrix metalloproteinase MMP-2 in the induction of TGF-β1 expression was significantly increasedIn the human lung sarcomatoid carcinoma paraffin has initially confirmed cases of lung sarcomatoid carcinoma have VE-cadherin, MMP-2 protein expression in VM, including high expression of MMP-2 illustrates the strong invasion of pulmonary sarcomatoid carcinoma, poor prognosis, 5-year survival rate characteristics. Western Blot detected lung cancer cell by TGF-β1-induced key protein of vascular mimicry, VE-cadherin, showed high expression; zymography experiments confirmed MMP-2 after 15 d induction of SK-MES-1 cells compared with the control group has increased activity.Conclusions: 1)Pulmonary sarcomatoid carcinoma occurs EMT, EMT phenotype which may be related to TGF-β activation of the transcription factor Twist1 promote EMT occurrence. 2) TGF-β1 and vascular mimicry related to the promotion of the formation of pulmonary sarcomatoid carcinoma VM. 3) Through TGF-β1-induced, aggressive and athletic ability of SK-MES-1 is enhanced. 4)TGF-β1 promotes VE-cadherin, the critical protein of VM, upregulated. 5)TGF-β1-induced EMT in lung cancer cells could occur and form VM. |
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