Objective: To investigate the expressions of Beta-catenin, E-cadherin and MMP-7 and their implications in ovarian epithelial tumor. Methods: Immunohistochemical staining with SP method was conducted to identify the ...Objective: To investigate the expressions of Beta-catenin, E-cadherin and MMP-7 and their implications in ovarian epithelial tumor. Methods: Immunohistochemical staining with SP method was conducted to identify the expressions of Beta-catenin, E-cadherin and MMP-7 in ovarian epithelial tumor in 66 cases. Results: The abnormal expression rate of Beta-catenin in malignant ovarian epithelial tumor was higher than those in borderline and benign epithelial tumors (P〈0.05). The positive rates of E-cadherin in benign and borderline ovarian epithelial tumors were significantly greater than that in malignant epithelial tumor. The expression rates of MMP-7 in malignant and borderline ovarian epithelial tumors were higher than that in benign epithelial tumor (P〈0.05). Conclusion: The abnormal expressions of Beta-catenin, E-cadherin and MMP-7 might be used to indicate the malignance transform of ovarian epithelial tumors, but they have no significant correlation with peritoneal dropsy invasion, caul invasion and appendant invasion in ovarian epithelial tumor.展开更多
Summary: To investigate whether the change of E-cadherin (ECD) expression plays a role in the injury and repair of airway epithelial cells (AEC) caused by smoking, porcine AECs were cultured by using an enzyme-dispers...Summary: To investigate whether the change of E-cadherin (ECD) expression plays a role in the injury and repair of airway epithelial cells (AEC) caused by smoking, porcine AECs were cultured by using an enzyme-dispersed method. After exposure of the AECs to cigarette smoke extract (CSE), the ECD expression in the cells was detected by using immunocytochemistry and in situ hybridization. The results showed that ECD was distributed on the plasma membrane at the cell junctions of AECs. After exposure to 20 % CSE, the membranous ECD expression was decreased, the cytoplasmic ECD expression was increased (P<0.01) as the exposure time went on. But the content of ECD mRNA in the AECs did not chang. It suggests that the change of ECD ex- pression is regulated at the posttranslational level and plays a role in the injury and repair of AEC caused by smoking.展开更多
AIM: To analyze the relationship between clinical features and epithelial mesenchymal transition (EMT) in retinoblastoma (RB), further to investigate whether miR-200c regulates the EMT and migration of RB cells. ...AIM: To analyze the relationship between clinical features and epithelial mesenchymal transition (EMT) in retinoblastoma (RB), further to investigate whether miR-200c regulates the EMT and migration of RB cells. METHODS: Expression of EMT-related markers and tumor- related factors were detected by immuno-histochemistry analysis in RB tissue from 29 cases. Correlations between their expression and clinical characteristics were analyzed. The regulation effects of miR-200c on EMT-related markers, tumor-related factors were observed in mRNA level and protein level by real-time polymerase chain reaction (PCR) and Western blot, respectively, in Y79 and Weri-rbl cells. Its effects on migration force of these RB cell lines were also detected with Transwell test. RESULTS: Lower expression of E-cadherin was present in the cases with malignant prognosis. MiR-200c promoted the expression of E-cadherin and decreased the expression of Vimentin and N-cadherin in Y79 and Weri-rbl cells. Migration force of RB cells could be inhibited by miR-200c. CONCLUSION: EMT might be associated with bad prognosis in RB. MiR-200c suppresses the migration of retinoblastomatous cells by reverse EMT.展开更多
The aim of this study is to reveal the regulation mechanism of the effect of Semen vaccariae and Taraxacu mogono on the cell-cell adhersion molecule, E-cadherin and β-catenin on the proliferation role and secretion f...The aim of this study is to reveal the regulation mechanism of the effect of Semen vaccariae and Taraxacu mogono on the cell-cell adhersion molecule, E-cadherin and β-catenin on the proliferation role and secretion function of bovine mammary epithelial cells cultured in vitro. Firstly, the epithelial character of bovine mammary epithelial cells was authenticated using immunofluorescence, then the cell grow curve was observed and investigated after S. vaccariae and T. mogono treatment. On the effect of S. vaccariae and T. mogono, cell adhesion molecules E-cadherin, β-catenin and CycinD1 mRNA and protein were detected by qRT-PCR and Western blotting, respectively. The results showed that the cellular keratin 18 expressed positively and proliferfated vigorously after S. vaccariae and T. mogono treament. The mRNA and protein levels of E-cadherin and CycinD1 were remarkably higher (P〈0.05) in 36 h after S. vaccariae and T. mogono treatment. The cell proliferation at 36 h was increased significantly (P〈0.05). In conclusion, S. vaccariae and T. mogono have a positive impact on the cell proliferation and an effect on the adhesion molecules E-cadherin, β-catenin and CycinD1 in the Wnt signaling pathway.展开更多
AIM:To investigate the role of suppressor of cytokine signaling 3 (SOCS3) silencing in epithelial-mesenchymal transition (EMT) involved in a human hepatocellular carcinoma MHCC97H cell line.METHODS:MHCC97H cells were ...AIM:To investigate the role of suppressor of cytokine signaling 3 (SOCS3) silencing in epithelial-mesenchymal transition (EMT) involved in a human hepatocellular carcinoma MHCC97H cell line.METHODS:MHCC97H cells were transiently transfected with SOCS3 small-interfering RNA (siRNA).Morphological changes of the transfected cells were observed under microscope.Expressions of E-cadherin,Vimentinand α-smooth muscle actin (α-SMA) were identified with immunofluorescence.Furthermore,protein expressions and mRNA levels of characteristic markers of EMT (E-cadherin,Vimentin,α-SMA and Snail) were detected by Western blotting,quantitative real-time polymerase chain reaction.Transforming growth factor-β1 (TGF-β1) levels in the supernatant were measured with enzyme-linked immunosorbent assay.RESULTS:The transfected cells with SOCS3 siRNA showed a morphological alteration from a typical cobblestone morphology to mesenchymal spindle-shaped and fusiform features.SOCS3 siRNA lessened immunofluorescent expression of E-cadherin,but elicited immunofluorescent expressions of Vimentin and α-SMA in MHCC97H cells.More importantly,compared with the negative control,depletion of SOCS3 resulted in the decrease of the epithelial marker E-cadherin (P < 0.05),and the increase of the mesenchymal markers Vimentin and α-SMA and the transcription factor Snail in MHCC97H cells (P < 0.05).Moreover,compared with the negative control,SOCS3 siRNA evidently enhanced TGF-β1 secretion in MHCC97H cells (200.20 ± 29.02 pg/mL vs 490.20 ± 92.43 pg/mL,P < 0.05).CONCLUSION:SOCS3 silencing is able to promote EMT in MHCC97H cells via changing the phenotypic characteristics and modulating the characteristic markers.展开更多
AIMTo investigate the expression of transcription factors Slug in human lens epithelial cells (HLECs) undergoing epithelial-mesenchymal transition (EMT) induced by connective tissue growth factor (CTGF).METHODSHLECs w...AIMTo investigate the expression of transcription factors Slug in human lens epithelial cells (HLECs) undergoing epithelial-mesenchymal transition (EMT) induced by connective tissue growth factor (CTGF).METHODSHLECs were treated with CTGF of different concentrations (20, 50 and 100 ng/mL) or without CTGF (control) for 24h. The morphological changes of HLECs were analysed by microscopy. The expression and cellular localization of Slug was evaluated by immumo-fluorescence. Expressions of Slug, E-cadherin and alpha smooth muscle actin (α-SMA) were further determined by Western blot analysis.RESULTSHLECs showed spidle fibrolasts-like characteristics and loosely connected each other after CTGF treatment. The immuno-fluorescence staining indicated that Slug was localized in the nuclei and its expression was induced by CTGF. The relative expressions of Slug protein were 1.64±0.11, 1.96 ±0.03, 3.12 ±0.10, and 4.08±0.14, respectively, in response to control group and treatment with CTGF of 20, 50 and 100 ng/mL (F=443.86, P<0.01). The increased Slug protein levels were correlated well with up-expression of α-SMA (0.78±0.05, 0.85±0.06, 2.17±0.15, 2.86±0.10; F=449.85, P<0.01) and down-expression of E-cadherin (2.50±0.11, 1.79±0.26, 1.05±0.14, 0.63±0.08; F=101.55, P<0.01).CONCLUSIONTranscription factor Slug may be involved in EMT of HLECs induced by CTGF in vitro.展开更多
目的:探究益肾通癃汤对前列腺癌(PCa)皮下种植瘤模型裸鼠N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)表达的影响。方法:用前列腺癌PC-3细胞悬液建立PCa皮下种植瘤模型裸鼠,造模成功后将其随机分为模型组、益肾通癃汤低剂量组、益肾通...目的:探究益肾通癃汤对前列腺癌(PCa)皮下种植瘤模型裸鼠N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)表达的影响。方法:用前列腺癌PC-3细胞悬液建立PCa皮下种植瘤模型裸鼠,造模成功后将其随机分为模型组、益肾通癃汤低剂量组、益肾通癃汤中剂量组、益肾通癃汤高剂量组,每组10只。各给药组灌胃给予相应药物,模型组灌胃给予等体积生理盐水,2次/d,连续3周。给药结束后,观察裸鼠一般状态,测量体质量及瘤体大小,称重并计算抑瘤率;运用HE染色观察各组裸鼠种植瘤病理学改变;ELISA法检测各组裸鼠血清中N-cadherin、Vimentin的水平;Western blotting检测各组裸鼠种植瘤组织中N-cadherin、Vimentin蛋白表达情况;PCR检测各组裸鼠种植瘤组织中N-cadherin m RNA、Vimentin m RNA表达情况。结果:与模型组比较,益肾通癃汤低、中、高剂量组裸鼠状态均可,瘤体质量,血清中N-cadherin、Vimentin水平,种植瘤组织中N-cadherin、Vimentin的蛋白及m RNA相对表达量均降低(P<0.05),瘤体细胞少量坏死;与益肾通癃汤低剂量组比较,益肾通癃汤中、高剂量组裸鼠一般状态可,瘤体质量,血清中N-cadherin、Vimentin水平、种植瘤组织中N-cadherin、Vimentin蛋白及mRNA相对表达量均降低(P<0.05),瘤体细胞部分坏死;与益肾通癃汤中剂量组比较,益肾通癃汤高剂量组裸鼠一般状态良好,瘤体质量,血清中N-cadherin水平,种植瘤组织中N-cadherin、Vimentin蛋白相对表达量,种植瘤组织中Vimentin mRNA相对表达量均降低(P<0.05),瘤体细胞坏死多;益肾通癃汤高剂量组裸鼠血清中Vimentin水平、种植瘤组织中N-cadherin mRNA相对表达量与益肾通癃汤中剂量组比较,差异无统计学意义(P>0.05);益肾通癃汤低、中、高剂量组裸鼠抑瘤率分别为26.67%、42.45%、66.26%。结论:益肾通癃汤对人前列腺癌PC-3细胞皮下种植瘤模型裸鼠N-cadherin、Vimentin蛋白的表达具有一定的抑制作用,其机制可能与抑制肿瘤上皮间质转化(EMT)过程相关。展开更多
AIM: To analyze the effect of three-dimensional (3D)-arrangement on the expression of epithelial-to-mesenchymal transition markers in pancreatic adenocarcinoma (PDAC) cells.METHODS: HPAF-II, HPAC, and PL45 PDAC cells ...AIM: To analyze the effect of three-dimensional (3D)-arrangement on the expression of epithelial-to-mesenchymal transition markers in pancreatic adenocarcinoma (PDAC) cells.METHODS: HPAF-II, HPAC, and PL45 PDAC cells were cultured in either 2D-monolayers or 3D-spheroids. Ultrastructure was analyzed by transmission electron microscopy. The expression of E-cadherin, β-catenin, N-cadherin, collagen type I (COL-I), vimentin, α-smooth muscle actin (αSMA), and podoplanin was assayed by confocal microscopy in cells cultured on 12-mm diameter round coverslips and in 3D-spheroids. Gene expression for E-cadherin, Snail, Slug, Twist, Zeb1, and Zeb2 was quantified by real-time PCR. E-cadherin protein level and its electrophoretic pattern were studied by Western blot in cell lysates obtained from cells grown in 2D-monolayers and 3D-spheroids.RESULTS: The E-cadherin/β-catenin complex was expressed in a similar way in plasma membrane cell boundaries in both 2D-monolayers and 3D-spheroids. E-cadherin increased in lysates obtained from 3D-spheroids, while cleavage fragments were more evident in 2D-monolayers. N-cadherin expression was observed in very few PDAC cells grown in 2D-monolayers, but was more evident in 3D-spheroids. Some cells expressing COL-I were observed in 3D-spheroids. Podoplanin, expressed in collectively migrating cells, and αSMA were similarly expressed in both experimental conditions. The concomitant maintenance of the E-cadherin/β-catenin complex at cell boundaries supports the hypothesis of a collective migration for these cells, which is consistent with podoplanin expression.CONCLUSION: We show that a 3D-cell culture model could provide deeper insight into understanding the biology of PDAC and allow for the detection of marked differences in the phenotype of PDAC cells grown in 3D-spheroids.展开更多
Breast cancer stem cells (BCSCs) are a small subpopulation of cancer cells having the ability of self-renewing and multi-lineage differentiation, which have also been termed as “tumor-initiating cells”. And in recen...Breast cancer stem cells (BCSCs) are a small subpopulation of cancer cells having the ability of self-renewing and multi-lineage differentiation, which have also been termed as “tumor-initiating cells”. And in recent years, the role of epithelial mesenchymal transition (EMT) in malignant tumors has been valued. This paper will briefly review and discuss the relationship between BCSCs and EMT.展开更多
基金the Scientific Research Start Found of Chongqing Medical University (No.QD200201)Project of Chongqing Science and Technology Committee (No.040307)
文摘Objective: To investigate the expressions of Beta-catenin, E-cadherin and MMP-7 and their implications in ovarian epithelial tumor. Methods: Immunohistochemical staining with SP method was conducted to identify the expressions of Beta-catenin, E-cadherin and MMP-7 in ovarian epithelial tumor in 66 cases. Results: The abnormal expression rate of Beta-catenin in malignant ovarian epithelial tumor was higher than those in borderline and benign epithelial tumors (P〈0.05). The positive rates of E-cadherin in benign and borderline ovarian epithelial tumors were significantly greater than that in malignant epithelial tumor. The expression rates of MMP-7 in malignant and borderline ovarian epithelial tumors were higher than that in benign epithelial tumor (P〈0.05). Conclusion: The abnormal expressions of Beta-catenin, E-cadherin and MMP-7 might be used to indicate the malignance transform of ovarian epithelial tumors, but they have no significant correlation with peritoneal dropsy invasion, caul invasion and appendant invasion in ovarian epithelial tumor.
基金This project was supported by a grant from National Natural Science Foundation of China !(39570288).
文摘Summary: To investigate whether the change of E-cadherin (ECD) expression plays a role in the injury and repair of airway epithelial cells (AEC) caused by smoking, porcine AECs were cultured by using an enzyme-dispersed method. After exposure of the AECs to cigarette smoke extract (CSE), the ECD expression in the cells was detected by using immunocytochemistry and in situ hybridization. The results showed that ECD was distributed on the plasma membrane at the cell junctions of AECs. After exposure to 20 % CSE, the membranous ECD expression was decreased, the cytoplasmic ECD expression was increased (P<0.01) as the exposure time went on. But the content of ECD mRNA in the AECs did not chang. It suggests that the change of ECD ex- pression is regulated at the posttranslational level and plays a role in the injury and repair of AEC caused by smoking.
基金Supported by the National Natural Science Foundation of China(No.81072221)National Science Foundation of Hunan Province(No.14JJ2005)
文摘AIM: To analyze the relationship between clinical features and epithelial mesenchymal transition (EMT) in retinoblastoma (RB), further to investigate whether miR-200c regulates the EMT and migration of RB cells. METHODS: Expression of EMT-related markers and tumor- related factors were detected by immuno-histochemistry analysis in RB tissue from 29 cases. Correlations between their expression and clinical characteristics were analyzed. The regulation effects of miR-200c on EMT-related markers, tumor-related factors were observed in mRNA level and protein level by real-time polymerase chain reaction (PCR) and Western blot, respectively, in Y79 and Weri-rbl cells. Its effects on migration force of these RB cell lines were also detected with Transwell test. RESULTS: Lower expression of E-cadherin was present in the cases with malignant prognosis. MiR-200c promoted the expression of E-cadherin and decreased the expression of Vimentin and N-cadherin in Y79 and Weri-rbl cells. Migration force of RB cells could be inhibited by miR-200c. CONCLUSION: EMT might be associated with bad prognosis in RB. MiR-200c suppresses the migration of retinoblastomatous cells by reverse EMT.
基金supported by the National Basic Research Program of China(2011CB100804)
文摘The aim of this study is to reveal the regulation mechanism of the effect of Semen vaccariae and Taraxacu mogono on the cell-cell adhersion molecule, E-cadherin and β-catenin on the proliferation role and secretion function of bovine mammary epithelial cells cultured in vitro. Firstly, the epithelial character of bovine mammary epithelial cells was authenticated using immunofluorescence, then the cell grow curve was observed and investigated after S. vaccariae and T. mogono treatment. On the effect of S. vaccariae and T. mogono, cell adhesion molecules E-cadherin, β-catenin and CycinD1 mRNA and protein were detected by qRT-PCR and Western blotting, respectively. The results showed that the cellular keratin 18 expressed positively and proliferfated vigorously after S. vaccariae and T. mogono treament. The mRNA and protein levels of E-cadherin and CycinD1 were remarkably higher (P〈0.05) in 36 h after S. vaccariae and T. mogono treatment. The cell proliferation at 36 h was increased significantly (P〈0.05). In conclusion, S. vaccariae and T. mogono have a positive impact on the cell proliferation and an effect on the adhesion molecules E-cadherin, β-catenin and CycinD1 in the Wnt signaling pathway.
基金Supported by Program for Changjiang Scholars and Innovative Research Team in Universities,PCSIRT No.1171National Natural Science Foundation of China,No.81201925 and No.81001588+1 种基金Specialized Research Fund of the Second Affiliated Hospital of Xi'an Jiaotong University School of Medicine of China,No.RC(XM)201108the Fundamental Research Funds for the Central Universities,No.08143048
文摘AIM:To investigate the role of suppressor of cytokine signaling 3 (SOCS3) silencing in epithelial-mesenchymal transition (EMT) involved in a human hepatocellular carcinoma MHCC97H cell line.METHODS:MHCC97H cells were transiently transfected with SOCS3 small-interfering RNA (siRNA).Morphological changes of the transfected cells were observed under microscope.Expressions of E-cadherin,Vimentinand α-smooth muscle actin (α-SMA) were identified with immunofluorescence.Furthermore,protein expressions and mRNA levels of characteristic markers of EMT (E-cadherin,Vimentin,α-SMA and Snail) were detected by Western blotting,quantitative real-time polymerase chain reaction.Transforming growth factor-β1 (TGF-β1) levels in the supernatant were measured with enzyme-linked immunosorbent assay.RESULTS:The transfected cells with SOCS3 siRNA showed a morphological alteration from a typical cobblestone morphology to mesenchymal spindle-shaped and fusiform features.SOCS3 siRNA lessened immunofluorescent expression of E-cadherin,but elicited immunofluorescent expressions of Vimentin and α-SMA in MHCC97H cells.More importantly,compared with the negative control,depletion of SOCS3 resulted in the decrease of the epithelial marker E-cadherin (P < 0.05),and the increase of the mesenchymal markers Vimentin and α-SMA and the transcription factor Snail in MHCC97H cells (P < 0.05).Moreover,compared with the negative control,SOCS3 siRNA evidently enhanced TGF-β1 secretion in MHCC97H cells (200.20 ± 29.02 pg/mL vs 490.20 ± 92.43 pg/mL,P < 0.05).CONCLUSION:SOCS3 silencing is able to promote EMT in MHCC97H cells via changing the phenotypic characteristics and modulating the characteristic markers.
基金Supported by National Natural Science Foundation of China(No.81470614,No.81460163,No.81300786)Fundamental Research Funds for the Central Universities(No.xjj2014146)+1 种基金Specialized Research Fund for the Doctoral Program of Higher Education(No.20133601120012)Key International Communication Project of Shaanxi province(No.2012KW-31)
文摘AIMTo investigate the expression of transcription factors Slug in human lens epithelial cells (HLECs) undergoing epithelial-mesenchymal transition (EMT) induced by connective tissue growth factor (CTGF).METHODSHLECs were treated with CTGF of different concentrations (20, 50 and 100 ng/mL) or without CTGF (control) for 24h. The morphological changes of HLECs were analysed by microscopy. The expression and cellular localization of Slug was evaluated by immumo-fluorescence. Expressions of Slug, E-cadherin and alpha smooth muscle actin (α-SMA) were further determined by Western blot analysis.RESULTSHLECs showed spidle fibrolasts-like characteristics and loosely connected each other after CTGF treatment. The immuno-fluorescence staining indicated that Slug was localized in the nuclei and its expression was induced by CTGF. The relative expressions of Slug protein were 1.64±0.11, 1.96 ±0.03, 3.12 ±0.10, and 4.08±0.14, respectively, in response to control group and treatment with CTGF of 20, 50 and 100 ng/mL (F=443.86, P<0.01). The increased Slug protein levels were correlated well with up-expression of α-SMA (0.78±0.05, 0.85±0.06, 2.17±0.15, 2.86±0.10; F=449.85, P<0.01) and down-expression of E-cadherin (2.50±0.11, 1.79±0.26, 1.05±0.14, 0.63±0.08; F=101.55, P<0.01).CONCLUSIONTranscription factor Slug may be involved in EMT of HLECs induced by CTGF in vitro.
文摘目的:探究益肾通癃汤对前列腺癌(PCa)皮下种植瘤模型裸鼠N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)表达的影响。方法:用前列腺癌PC-3细胞悬液建立PCa皮下种植瘤模型裸鼠,造模成功后将其随机分为模型组、益肾通癃汤低剂量组、益肾通癃汤中剂量组、益肾通癃汤高剂量组,每组10只。各给药组灌胃给予相应药物,模型组灌胃给予等体积生理盐水,2次/d,连续3周。给药结束后,观察裸鼠一般状态,测量体质量及瘤体大小,称重并计算抑瘤率;运用HE染色观察各组裸鼠种植瘤病理学改变;ELISA法检测各组裸鼠血清中N-cadherin、Vimentin的水平;Western blotting检测各组裸鼠种植瘤组织中N-cadherin、Vimentin蛋白表达情况;PCR检测各组裸鼠种植瘤组织中N-cadherin m RNA、Vimentin m RNA表达情况。结果:与模型组比较,益肾通癃汤低、中、高剂量组裸鼠状态均可,瘤体质量,血清中N-cadherin、Vimentin水平,种植瘤组织中N-cadherin、Vimentin的蛋白及m RNA相对表达量均降低(P<0.05),瘤体细胞少量坏死;与益肾通癃汤低剂量组比较,益肾通癃汤中、高剂量组裸鼠一般状态可,瘤体质量,血清中N-cadherin、Vimentin水平、种植瘤组织中N-cadherin、Vimentin蛋白及mRNA相对表达量均降低(P<0.05),瘤体细胞部分坏死;与益肾通癃汤中剂量组比较,益肾通癃汤高剂量组裸鼠一般状态良好,瘤体质量,血清中N-cadherin水平,种植瘤组织中N-cadherin、Vimentin蛋白相对表达量,种植瘤组织中Vimentin mRNA相对表达量均降低(P<0.05),瘤体细胞坏死多;益肾通癃汤高剂量组裸鼠血清中Vimentin水平、种植瘤组织中N-cadherin mRNA相对表达量与益肾通癃汤中剂量组比较,差异无统计学意义(P>0.05);益肾通癃汤低、中、高剂量组裸鼠抑瘤率分别为26.67%、42.45%、66.26%。结论:益肾通癃汤对人前列腺癌PC-3细胞皮下种植瘤模型裸鼠N-cadherin、Vimentin蛋白的表达具有一定的抑制作用,其机制可能与抑制肿瘤上皮间质转化(EMT)过程相关。
基金Supported by the University of Milan(Project B grant) and core support grant from the Wellcome Trust and MRC to the Wellcome Trust-Medical Research Council Cambridge Stem Cell Institute
文摘AIM: To analyze the effect of three-dimensional (3D)-arrangement on the expression of epithelial-to-mesenchymal transition markers in pancreatic adenocarcinoma (PDAC) cells.METHODS: HPAF-II, HPAC, and PL45 PDAC cells were cultured in either 2D-monolayers or 3D-spheroids. Ultrastructure was analyzed by transmission electron microscopy. The expression of E-cadherin, β-catenin, N-cadherin, collagen type I (COL-I), vimentin, α-smooth muscle actin (αSMA), and podoplanin was assayed by confocal microscopy in cells cultured on 12-mm diameter round coverslips and in 3D-spheroids. Gene expression for E-cadherin, Snail, Slug, Twist, Zeb1, and Zeb2 was quantified by real-time PCR. E-cadherin protein level and its electrophoretic pattern were studied by Western blot in cell lysates obtained from cells grown in 2D-monolayers and 3D-spheroids.RESULTS: The E-cadherin/β-catenin complex was expressed in a similar way in plasma membrane cell boundaries in both 2D-monolayers and 3D-spheroids. E-cadherin increased in lysates obtained from 3D-spheroids, while cleavage fragments were more evident in 2D-monolayers. N-cadherin expression was observed in very few PDAC cells grown in 2D-monolayers, but was more evident in 3D-spheroids. Some cells expressing COL-I were observed in 3D-spheroids. Podoplanin, expressed in collectively migrating cells, and αSMA were similarly expressed in both experimental conditions. The concomitant maintenance of the E-cadherin/β-catenin complex at cell boundaries supports the hypothesis of a collective migration for these cells, which is consistent with podoplanin expression.CONCLUSION: We show that a 3D-cell culture model could provide deeper insight into understanding the biology of PDAC and allow for the detection of marked differences in the phenotype of PDAC cells grown in 3D-spheroids.
文摘Breast cancer stem cells (BCSCs) are a small subpopulation of cancer cells having the ability of self-renewing and multi-lineage differentiation, which have also been termed as “tumor-initiating cells”. And in recent years, the role of epithelial mesenchymal transition (EMT) in malignant tumors has been valued. This paper will briefly review and discuss the relationship between BCSCs and EMT.