Journal: Journal of Ovarian Research

Article Title: MicroRNA-200c overexpression inhibits tumorigenicity and metastasis of CD117 + CD44 + ovarian cancer stem cells by regulating epithelial-mesenchymal transition

doi: 10.1186/1757-2215-6-50

Figure Lengend Snippet: Identification, selection and detection of the miR-200c expression in the CD44 + CD117 + CSCs. A . The CD117 + CD44 + CSCs isolated from the human EOC SKOV3 cell line were identified by FCM; B . The miR-200c expression differences between the non CD117 + CD44 + CSCs and the CD117 + CD44 + CSCs were detected by qRT-PCR; C . The top and bottom panels show the morphological structures of the CD44 + CD117 + CSCs with the stable miR-200c overexpression under a fluorescence microscope and a light microscope, respectively. The CD44 + CD117 + CSCs transduced with lentivirus-mock (left) or lentivirus miR-200c (right) in the stem cell culture medium; D . miR-200c expression differences among the CD44 + CD117 + CSCs transduced with lentivirus-mock, the lentivirus miR-200c and without lentivirus infection were detected by qRT-PCR. E . E-cadherin was detected by IHC assay in tumors of mice injected with the different cells. The labels ‘WT, mock and miR-200c’ denote the CD44 + CD117 + CSCs, the CD44 + CD117 + CSCs transduced with lentivirus-mock, and the lentivirus miR-200c, respectively. These labels are also used in Figures , , below.

Article Snippet: Briefly, the CD44 + subsets were isolated by using the mouse antihuman CD44 antibody coupled to magnetic microbeads (Miltenyi Biotec., Germany); the resulting cells were then depleted of CD117 − subsets by using the mouse antihuman CD117 antibody coupled to magnetic microbeads (Miltenyi Biotec., Germany).

Techniques: Selection, Expressing, Isolation, Quantitative RT-PCR, Over Expression, Fluorescence, Microscopy, Light Microscopy, Transduction, Stem Cell Culture, Infection, Injection

Journal: Journal of Ovarian Research

Article Title: MicroRNA-200c overexpression inhibits tumorigenicity and metastasis of CD117 + CD44 + ovarian cancer stem cells by regulating epithelial-mesenchymal transition

doi: 10.1186/1757-2215-6-50

Figure Lengend Snippet: The CD44 + CD117 + CSCs transduced with lentivirus miR-200c reduced the ability of colony forming, cell migration, invasion and cell proliferation in vitro . A . The plate colony forming assay; B . The wound healing assay; C . The transwell invasive assay; D - F . The results of the statistical analysis for the plate colony forming, the wound healing and the transwell invasion, respectively. G : The cell proliferation ability of the different cells in vitro was detected by MTT assay. * p < 0.05 and ** p < 0.01.

Article Snippet: Briefly, the CD44 + subsets were isolated by using the mouse antihuman CD44 antibody coupled to magnetic microbeads (Miltenyi Biotec., Germany); the resulting cells were then depleted of CD117 − subsets by using the mouse antihuman CD117 antibody coupled to magnetic microbeads (Miltenyi Biotec., Germany).

Techniques: Transduction, Migration, In Vitro, Wound Healing Assay, MTT Assay

Journal: Journal of Ovarian Research

Article Title: MicroRNA-200c overexpression inhibits tumorigenicity and metastasis of CD117 + CD44 + ovarian cancer stem cells by regulating epithelial-mesenchymal transition

doi: 10.1186/1757-2215-6-50

Figure Lengend Snippet: The miR-200c overexpression decreased tumor progression of CD44 + CD117 + CSCs in xenograft mice. A . 5 × 10 4 CD44 + CD117 + CSCs transduced with lentivirus miR-200c or lentivirus-mock or without infection were subcutaneously injected in the nude mice ( n = 6/group). The figure shows the tumor growth dynamic state in the mice injected with the different CSCs (line, mean ± SE). B . Images of the tumor tissues dissected from the mice on day 56 after injection. C . Tumor-free mice on day 56 after injection(line, mean ± SE). * p < 0.01 and ** p < 0.01.

Article Snippet: Briefly, the CD44 + subsets were isolated by using the mouse antihuman CD44 antibody coupled to magnetic microbeads (Miltenyi Biotec., Germany); the resulting cells were then depleted of CD117 − subsets by using the mouse antihuman CD117 antibody coupled to magnetic microbeads (Miltenyi Biotec., Germany).

Techniques: Over Expression, Transduction, Infection, Injection

Journal: Journal of Ovarian Research

Article Title: MicroRNA-200c overexpression inhibits tumorigenicity and metastasis of CD117 + CD44 + ovarian cancer stem cells by regulating epithelial-mesenchymal transition

doi: 10.1186/1757-2215-6-50

Figure Lengend Snippet: Impacts of the overexpression of miR-200c on ZEB1, E-Cadherin, and the Vimentin expressions in tumors, and tumor cell metastasis in mouse lungs. A - C . The qRT-PCR analysis results of the mRNA expression of ZEB1, Vimentin and E-Cadherin in the tumor tissues of EOC-bearing nude mice after injection with CD44 + CD117 + CSCs. D . The protein expression of ZEB1, E-Cadherin and Vimentin from the Western blotting analysis. E - G . The protein expression results from the gradation scanning analysis. H . The tumor metastasis in mouse lungs (stained in hematoxylin and eosin). I . The statistically significant differences are indicated by asterisk for ** p < 0.01.

Article Snippet: Briefly, the CD44 + subsets were isolated by using the mouse antihuman CD44 antibody coupled to magnetic microbeads (Miltenyi Biotec., Germany); the resulting cells were then depleted of CD117 − subsets by using the mouse antihuman CD117 antibody coupled to magnetic microbeads (Miltenyi Biotec., Germany).

Techniques: Over Expression, Quantitative RT-PCR, Expressing, Injection, Western Blot, Staining

Journal: Journal of Ovarian Research

Article Title: MicroRNA-200c overexpression inhibits tumorigenicity and metastasis of CD117 + CD44 + ovarian cancer stem cells by regulating epithelial-mesenchymal transition

doi: 10.1186/1757-2215-6-50

Figure Lengend Snippet: Down-regulation of ZEB1 inhibited the colony forming, cell migration and invasion in vitro . A - C . The representative images from the results of the plate clone forming assay, the wound healing assay, and the transwell invasive assay, respectively; D - F give the between-group statistical differences in the plate colony forming ratio, in the healing degree, and in the invasive cells, respectively. The labels ‘WT, scramble and shZEB1’ denote the CD44 + CD117 + CSCs, the CD44 + CD117 + CSCs stably transfected with scramble shRNA, and the CD44 + CD117 + CSCs stably transfected with shZEB1, respectively. * p < 0.05 and ** p < 0.01. These labels are also used in Figure below.

Article Snippet: Briefly, the CD44 + subsets were isolated by using the mouse antihuman CD44 antibody coupled to magnetic microbeads (Miltenyi Biotec., Germany); the resulting cells were then depleted of CD117 − subsets by using the mouse antihuman CD117 antibody coupled to magnetic microbeads (Miltenyi Biotec., Germany).

Techniques: Migration, In Vitro, Wound Healing Assay, Stable Transfection, Transfection, shRNA

Journal: Journal of Ovarian Research

Article Title: MicroRNA-200c overexpression inhibits tumorigenicity and metastasis of CD117 + CD44 + ovarian cancer stem cells by regulating epithelial-mesenchymal transition

doi: 10.1186/1757-2215-6-50

Figure Lengend Snippet: Down-regulation of ZEB1 reduced tumor growth and metastasis in vivo xenograft mouse. A - B . The tumor dynamic growth curves and tumor bearing mice’ survival. C . The photos of the tumor tissues dissected from the mice on day 56 after injection. D . The protein expression results from the Western blotting analysis. E - G . The protein expression results from gradation scanning analysis. H . The HE staining results from the tumor tissue sections at 400× magnification; The tumor cell metastases were visible in the samples of the CD44 + CD117 + WT(+++) and the CD44 + CD117 + scramble (+++); a few tumor cells were seen in the lungs of the nude mice injected with the CD44 + CD117 + shZEB1 (+). I.The statistical analysis results of the metastatic tumor cells in the lungs of the mice injected with the different CSCs. * p < 0.05 and ** p < 0.01.

Article Snippet: Briefly, the CD44 + subsets were isolated by using the mouse antihuman CD44 antibody coupled to magnetic microbeads (Miltenyi Biotec., Germany); the resulting cells were then depleted of CD117 − subsets by using the mouse antihuman CD117 antibody coupled to magnetic microbeads (Miltenyi Biotec., Germany).

Techniques: In Vivo, Injection, Expressing, Western Blot, Staining