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@#[Abstract] Objective: To investigate the effect of RG108 on the proliferation and apoptosis of human non-small cell lung cancer (NSCLC) cell lines (A549, H1299) and explore its molecular mechanism. Methods: A549 and H1299 cells were cultured in vitro and treated with different concentrations of RG108. The cell proliferation, cell cycle and apoptosis were detected by MTT assay and Flow cytometry, respectively. qPCR and Western blotting (WB) were used to detect the TFPI-2 mRNA and protein expressions as well as the expression of TMPRSS4 in cells. Meanwhile, the methylation status and degree of TFPI-2 promoter in cells were detected with Methylation-specific PCR (MSP) and colorimetry. Finally, siRNA-TFPI-2 and pcDNA3.0-TMPRSS4 plasmids were used to silence TFPI-2 or overexpress TMPRSS4, and then the changes in cell proliferation and apoptosis were detected. Results: After treatment with RG108, the proliferation rate of A549 and H1299 cells were significantly decreased (all P<0.05), while the apoptosis rate were significantly increased(P<0.05), the cell cycle were arrested in G1/S phase (P<0.05), and the intracellular mRNA and protein expressions of TFPI-2 were significantly increased (P<0.01 or P<0.05). Meanwhile, the methylase degree in TFPI-2 promoter region and the expression of TMPRSS4 in cells were all significanly decreased ( all P<0.05). After TFPI-2 silence, the proliferation levels of A549 and H1299 cells were significantly increased(all P<0.05); however, the apoptosis rate of A549 and H1299 cells were significantly reduced after transfection with pcDNA3.0-TMPRSS4(all P<0.05). Conclusion: RG108 can inhibit proliferation of A549 and H1299 cells and promote apoptosis by inhibiting the methylation of TFPI-2 and negatively regulates the expression of TMPRSS4.
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Objective To explore a newly discovered transmembrane protease serine 4 (TMPRSS4) isoforms and its molecular charcteristics in transfected colon cancer cells. Methods The named T4-1A and T4-1B of the TMPRSS4-isoforms were authenticated by the RT-PCR and Western blot,and then transfected to human colon cancer cells (DLD-1). Those stable transfected cells of migration and invasion were illustrated using wound healing assays and matrigel invasion assays. Results Successfully constructed T4-1A and T4-1B recombinant vectors,and obtained T4-1A and T4-1B transfected cell lines,and their T4-1A and T4-1B were highly expressed in stable DLD-1. Compared to cells transfected with empty vector of pcDNA6,the transfected DLD-1 with T4-1A enhanced migration and invasion were statistical significance (P < 0. 05). However,compared to pcDNA6 no significant1 difference was found for T4-1B. Moreover,the biological characteristics of T4-1A and TMPRSS4 were very similar. Conclusion The T4-1A and T4-1B is newly TMPRSS4-isoforms,and protease domain included to a T4-1A has further facilitated the migration and invasion of colon cancer cells,and further studies provide a theoretic base in the molecular biomedical characteristics of TMPRSS4.
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Objective To investigate the relationship between the expressions of TMPRSS4 and clinical pathologi-cal parameters in gastric carcinoma. To explore its significance in judging the prognosis of patients. To analyze the expression of TMPRSS4 in the epithelial-mesenchymal transition. Methods The expressions of TMPRSS4, E-cad-herin and Vimentin were detected by immunohistochemistry ( SABC) in 100 cases of gastric carcinoma specimens and corresponding adjacent normal tissue. Results In gastric carcinoma, TMPRSS4, E-cadherin and Vimentin positive rate was 47%,45%,37%. In adjacent tissues,their positive rate was 9%,30%,4%. The differences be-tween tumor tissues and adjacent tissues had statistical significance ( P0.05). The 5-year survival rate of patients with high TMPRSS4 expression was significantly lower than that in patients with low expression. The high TMPRSS4 expression was significantly correlated in gastric carcinoma accompanied by low expression of E-cadherin (rs = -0.207,P=0.038) and high Vimentin expression (rs=0.233,P=0.020). Conclusion The expression of TMPRSS4 is closely related to the biological characteristics in gastric carcinoma,detection the expression of TMPRSS4 is valuable in predicting tumor prognosis,invasion and metastasis. TMPRSS4 may promote invasion, metastasis of human gastric carcinoma through epithelial-mesenchymal transition by reduce the expession of E-cadherin.
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The epithelial-mesenchymal transition (EMT) is one mechanism by which cells with mesenchymal features can be generated and is a fundamental event in morphogenesis. Recently, invasion and metastasis of cancer cells from the primary tumor are now thought to be initiated by the developmental process termed the EMT, whereby epithelial cells lose cell polarity and cell-cell interactions, and gain mesenchymal phenotypes with increased migratory and invasive properties. The EMT is believed to be an important step in metastasis and is implicated in cancer progression, although the influence of the EMT in clinical specimens has been debated. This review presents the recent results of two cell surface proteins, the functions and underlying mechanisms of which have recently begun to be demonstrated, as novel regulators of the molecular networks that induce the EMT and cancer progression.
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Polaridad Celular , Células Epiteliales , Transición Epitelial-Mesenquimal , Proteínas de la Membrana , Membranas , Morfogénesis , Metástasis de la Neoplasia , FenotipoRESUMEN
Type Ⅱ transmembrane serine proteases 4 (TMPRSS4) is a novel type Ⅱ transmembrane serine protease.Present study showed that its expression was related with tumor invasion and metastasis,although its oncogenic significance and molecular mechanisms are still unknown.In this review,the author try to introduce its structure,biological function and mechanism in tumor invasion and metastasis.
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Objective To investigate the expression of TMPRSS4 mRNA,protein in human pancreatic cancer tissues and to explore the relationship between the expression of TMPRSS4 protein and the clinicopathologic parameters.Methods Real-time PCR and Western blotting were used to detect the expressions of TMPRSS4 mRNA and protein in 16 samples of pancreatic cancer tissues and adjacent normal pancreatic tissues.The expression of TMPRSS4 protein in 61 samples of pancreatic cancer tissues and 26 samples of adjacent pancreatic tissues and 4 samples of normal pancreatic tissues was detected by using immunohistochemistry and its relationship with clinicopathological features was analyzed.Results The expression of TMPRSS4 mRNA and protein of pancreatic cancer tissues were significantly higher than those in adjacent pancreatic tissues (9.09 ± 7.01 vs.1.27 ± 0.72; 1.223 ± 0.125 vs.0.667 ± 0.106,P ＜ 0.01 ) ;the expression rate of TMPRSS4 protein of pancreatic cancer tissues was 67.2％ (41/61),which were significantly higher than that in adjacent pancreatic tissues[3.8％ (1/26),P ＜ 0.01].There was no TMPRSS4 protein expression in normal pancreatic tissues.There was no significant correlation between the expression of TMPRSS4 protein and the age,gender,tumor location or tumor size was found.There was significant correlation between the expression of TMPRSS4 protein and the degree of differentiation,lymph node metastasis,and clinical staging (P ＜ 0.05 ).Conclusions TMPRSS4 protein is highly expressed in pancreatic cancer tissues,and the expression of TMPRSS4 is associated with the degree of malignancy of pancreatic cancer.
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Objective To study the influence of the small interfering RNA (siRNA) interference TMPRSS4 expression on human pancreatic cancer SW1990 cell's proliferation and invasion. Methods The four eukaryotic expression vector of TMPRSS4 gene were synthesized in vitro and were transfected transiently into human pancreatic cancer SW1990 cells. TMPRSS4 mRNA expression of transfected cells was detected by real-time RT-PCR. The most efficient eukaryotic expression vector was used to be transfected into SW1990 cells. By using G418, cell strain that can silence TMPRSS4 gene stably was screened. The TMPRSS4 mRNA expression of the stable cell strain was detected by real time PCR TMPRSS4 protein expression was detected by western blot. The proliferation ability of transfected SW1990 cells was detected by CCK-8 method. By Transwell, the invasion change of SW1990 cell was detected. Results A stable cell strain, SW1990/psi TMPRSS4, was successfully constructed, in which the expression level of TMPRSS4 could be reduced stably by RNA interference. Cell transfection efficiency was 82.9%. Compared with the control group, the TMPRSS4 mRNA and protein levels were reduced by 80.1% and 60% ,and number of penetrating cells was 118.6 ±13.4 in SW1990/psi TMPRSS4 group, which was significantly lower than those in the negative control group (157.4 ± 12.9) and control group (157.0±9.5, P <0.01). Cells invasion inhibitory rate was 24.5% in SW1990/psi TMPRSS4 group. The cell proliferation was not significantly different among all the groups. Conclusions A stable cell strain is screened successfully in which the expression level of TMPRSS4 can be reduced stably. The down-regulation of TMPRSS4 gene expression level can inhibit the invasion of SW1990 cells, but has no effect on cell proliferation.