Journal: Cancer Management and Research

Article Title: Overexpression of PDK4 is associated with cell proliferation, drug resistance and poor prognosis in ovarian cancer

doi: 10.2147/CMAR.S185015

Figure Lengend Snippet: PDK4 promotes cell proliferation and invasion. Notes: ( A ) Expression of PDK4 was determined by Western blot in ES2 transfected with PDK4 plasmid (left panel) or OVISE transfected with two different siRNAs (si-1, si-2, right panel). ( B ) MTT assay showed that PDK4 overexpression promoted proliferation in ES2 cells (left panel). PDK4 knockdown reduced proliferation in OVISE cells (right panel). The statistical analysis was performed between each group at 96-hour time point. ( C ) PDK4 overexpression upregulated invading ability of ES2 cells (upper panel). PDK4 knockdown decreased the invading ability of OVISE cells (lower panel) (* P <0.05). Magnification 200×.

Article Snippet: The human ovarian cancer cell lines COV504, OVISE, SKOV3, OV90, ES2 and OVCAR4 were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Western Blot, Transfection, Plasmid Preparation, MTT Assay, Over Expression, Knockdown

Journal: Cancer Management and Research

Article Title: Overexpression of PDK4 is associated with cell proliferation, drug resistance and poor prognosis in ovarian cancer

doi: 10.2147/CMAR.S185015

Figure Lengend Snippet: Overexpression of PDK4 confers ovarian cancer cells resistant to paclitaxel and cisplatin. Notes: Cell viability assay was performed in PDK4-overexpressed ES2 cells ( A ) and PDK4-knocdown OVISE cells ( B ) treated with different concentration of paclitaxel (Taxol) and cisplatin (Cis). ( C ) Overexpression of PDK4 decreased expression of cleaved caspase-3 which induced by Taxol as evaluated by Western blot and immunofluorescence. ( D ) Knocking down of PDK4 increased the expression of cleaved caspase-3 (bar, 100 µm). Paclitaxel-induced cell apoptosis was determined in ES2 cells transfected with PDK4 ( E ) and OVISE cells transfected with siRNAs ( F ) (* P <0.05).

Article Snippet: The human ovarian cancer cell lines COV504, OVISE, SKOV3, OV90, ES2 and OVCAR4 were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Over Expression, Viability Assay, Concentration Assay, Expressing, Western Blot, Immunofluorescence, Transfection

Journal: Cancer Management and Research

Article Title: Overexpression of PDK4 is associated with cell proliferation, drug resistance and poor prognosis in ovarian cancer

doi: 10.2147/CMAR.S185015

Figure Lengend Snippet: PDK4 enhances stemness and enriches CD133+ subpopulations of ovarian cancer cells. Notes: ( A ). Spheroid formation assay for ES2 cells transfected with PDK4 and OVISE cells transfected with siRNA. Representative images (left panel) and statistical measurement (right panel). ( B ) CD133+ subpopulations of aforementioned cells. Representative FACS profiles. ( C ) Stem cell-associated gene expression was measured by qRT-PCR in aforementioned cells (* P <0.05, ** P <0.01). Abbreviations: FACS, fluorescence-activated cell sorting; qRT-PCR, quantitative real-time PCR.

Article Snippet: The human ovarian cancer cell lines COV504, OVISE, SKOV3, OV90, ES2 and OVCAR4 were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Tube Formation Assay, Transfection, Gene Expression, Quantitative RT-PCR, Fluorescence, FACS, Real-time Polymerase Chain Reaction

Journal: Cancer Management and Research

Article Title: Overexpression of PDK4 is associated with cell proliferation, drug resistance and poor prognosis in ovarian cancer

doi: 10.2147/CMAR.S185015

Figure Lengend Snippet: The IC50 value of each group

Article Snippet: The human ovarian cancer cell lines COV504, OVISE, SKOV3, OV90, ES2 and OVCAR4 were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques:

Journal: Oncotarget

Article Title: Ovatodiolide targets chronic myeloid leukemia stem cells by epigenetically upregulating hsa-miR-155, suppressing the BCR-ABL fusion gene and dysregulating the PI3K/AKT/mTOR pathway

doi: 10.18632/oncotarget.23231

Figure Lengend Snippet: ( A ) Flow cytometry-based cell sorting of K562 cells stained with Isotype-PE, Isotype-FITC, CD38-PE, and CD34-FITC, into CD34 + 38 – and CD34 + 38 + populations. ( B ) Analysis of the protein expression levels of Bcr-abl, p-CrkL and p-Stat5 in CD34 + 38 − or CD34 + 38 + K562 cells, compared to control unsorted K562 cells. ( C ) Western blot analysis of the expression profile of Oct4, Sox2 and CD133 proteins in CD34 + 38 − or CD34 + 38 + K562 cells, compared to control unsorted K562 cells. ( D ) Relative mRNA expression levels of Oct4, Sox2, CD133 and Stat5 normalized to GAPDH, using RT-PCR. β-actin was used as loading control.

Article Snippet: The human CML cell line K562 was obtained from the American Type Culture Collection (ATCC.

Techniques: Flow Cytometry, FACS, Staining, Expressing, Control, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: Oncotarget

Article Title: Ovatodiolide targets chronic myeloid leukemia stem cells by epigenetically upregulating hsa-miR-155, suppressing the BCR-ABL fusion gene and dysregulating the PI3K/AKT/mTOR pathway

doi: 10.18632/oncotarget.23231

Figure Lengend Snippet: ( A ) The chemical structure of Imatinib, with molecular formula C29H31N7O and molar mass 493.603 g/mol. ( B ) The effect of 0.01–1.0 μM Imatinib on the cell viability of CD34 + 38 − or CD34 + 38 + K562 cells, compared to unsorted parental K562 cells. ( C ) Western blot analyses of the effect of 0.01–1.0 μM Imatinib on the protein expression levels of Bcr-abl, p-CrkL, Stat5, and MDR in CD34 + 38 − or CD34 + 38 + K562 cells. ( D ) The chemical structure of Ova, with molecular formula C20H24O4 and molar mass 328.408 g/mol. ( E ) The effect of 0.01–1.0 μM Imatinib and/or 2.5–10 μM Ova on the cell viability of CD34 + 38 − or CD34 + 38 + K562 cells. ( F ) Western blot analyses of the effect of 0.01–1.0 μM Imatinib and/or 2.5–10 μM Ova on the protein expression levels of Bcr-abl, p-CrkL, Stat5, and MDR in CD34 + 38 − or CD34 + 38 + K562 cells. β-actin was used as loading control. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The human CML cell line K562 was obtained from the American Type Culture Collection (ATCC.

Techniques: Western Blot, Expressing, Control

Journal: Oncotarget

Article Title: Ovatodiolide targets chronic myeloid leukemia stem cells by epigenetically upregulating hsa-miR-155, suppressing the BCR-ABL fusion gene and dysregulating the PI3K/AKT/mTOR pathway

doi: 10.18632/oncotarget.23231

Figure Lengend Snippet: ( A ) Tumorspehere formation assay for analysis of the effect of 1 μM Imatinib and/or 2.5 μM Ova treatment on the ability of CD34 + 38 − K562 cells to form tumorspheres. ( B ) Bar chart showing how treatment with 0.1–1 μM Imatinib and/or 2.5 μM Ova induce apoptosis in tumorspheres derived from CD34 + 38 − K562 cells. ( C ) Western blot analysis of Bcl-2 and Bax proteins expression level after treatment with 0.1–1 μM Imatinib and/or 2.5 μM Ova (left panel), and a graphical representation of the Bax/Bcl-2 ratio (right panel). ( D ) Western blot analysis of the effect of 0.1–1 μM Imatinib and/or 2.5 μM Ova on the expression levels of Bcr-abl, p-PI3K, p-Akt, Akt, p-mTOR, and CD44 proteins. β-actin was used as loading control. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The human CML cell line K562 was obtained from the American Type Culture Collection (ATCC.

Techniques: Tube Formation Assay, Derivative Assay, Western Blot, Expressing, Control

Journal: Oncotarget

Article Title: Ovatodiolide targets chronic myeloid leukemia stem cells by epigenetically upregulating hsa-miR-155, suppressing the BCR-ABL fusion gene and dysregulating the PI3K/AKT/mTOR pathway

doi: 10.18632/oncotarget.23231

Figure Lengend Snippet: ( A ) Comparative analysis of miR-155, miR-34a, miR-45-3p and miR-21 expression in CD34 + 38 − or parental K562 cells using RT-PCR. ( B ) The effect of Ova treatment on the expression of miR-155, miR-34a, miR-45-3p and miR-21 in CD34 + 38 – K562 cells, compared to the untreated CD34 + CD38 – or parental K562 counterparts. ( C ) Mapping of the 5’ UTR of miR-155 to the 3’ UTR of the PIK3CA by TargetScan (upper panel). The differential effect of exposure to miR-155-5p inhibitor or mimic on the Stat5, NF-kB, mTOR and PI3KCA mRNA level in CD34 + 38 – K562 cells, compared with control K562 cells (lower panel). ( D ) Western blot analysis of the changes in NF-κB, PIK3CA, mTOR, and Stat5 protein expression level in CD34 + 38 − K562 cells in response to miR-155 inhibitor or mimic, in comparison to the control group. β-actin was used as loading control. ( E ) RT-PCR analysis of the effect of 2.5 µM–5 µM Ova treatment on the expressions of hsa-miR-155, PIK3CA, mTOR, and STAT5 in CD34+ K562 cells, compared to their untreated counterparts. GAPDH was used as loading control. ( F ) Molecular docking to demonstrate direct interaction between chain B8 of hsa-miR-155 and chain A of PI3KCA with a docking score of -250.59, and a ligand rmsd of 178.10 Å. PIK3CA, in white color. hsa-miR-155 in golden yellow color. RMSD, root-mean-square deviation, is the measure of the average distance between the atoms (usually the backbone atoms) of superimposed molecules.

Article Snippet: The human CML cell line K562 was obtained from the American Type Culture Collection (ATCC.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Control, Western Blot, Comparison