Article Figures & Data
Figures
- Fig. 1.
Acquisition of oxaliplatin resistance induces morphologic changes consistent with EMT in CRC cells. KM12L4 (A) and HT29 (B) parental and OxR cells were assessed for morphologic changes consistent with EMT. Spindle-shaped cells with loss of polarity (red arrows), increased intercellular separation (green arrows), and pseudopodia (white arrows) were noted in the OxR cells but not in parental cells from both cell lines.
- Fig. 2.
OxR CRC cells have increased migratory and invasive capacity. Boyden chamber and modified Boyden chamber assays were done to compare the migratory and invasive capabilities of KM12L4 OxR and parental cells. A, at 48 hours, the OxR cells showed an ∼7.5-fold increase in the number of cells migrating through the collagen insert. B, also at 48 hours, the OxR cells exhibited an ∼15-fold increase in the number of cells invading through the Matrigel-coated collagen insert. HPF, high-power field; Ox, oxaliplatin.
- Fig. 3.
OxR CRC cells exhibit changes in localization of cellular EMT markers. Immunofluorescence staining for E-cadherin and β-catenin was done on KM12L4 (A) and HT29 (B) parental and OxR cells. OxR cells from both cell lines showed changes in localization of E-cadherin and β-catenin from their usual cell membrane-associated site. OxR cells exhibited E-cadherin in a disorganized cytoplasmic location and β-catenin was noted to translocate to the nucleus.
- Fig. 4.
OxR CRC cells exhibit molecular changes consistent with EMT. A, cell lysates and nuclear extracts from KM12L4 parental and OxR cells were subjected to Western blotting. Expression of the epithelial cellular adhesion molecules E-cadherin and plakoglobin was decreased in OxR cells compared with parental cells (top left). A marked increase in the expression of the mesenchymal marker vimentin was concurrently observed (top right). There was also increased expression of the EMT-related transcription factor Snail in nuclear extracts of OxR cells compared with parental cells (bottom). No change in expression of the EMT-related transcription factors Slug or Twist was noted. B, similar results were noted when validation studies were done in the HT29 parental and OxR cells (left) except that no up-regulation of mesenchymal markers was noted and no changes were noted in Snail, Slug, or Twist. However, nuclear expression of another transcription factor implicated in the induction of EMT, nuclear factor κB (NF-κB), was observed in the HT29 OxR cell line (right).
- Fig. 5.
OxR CRC cells have reduced cell number. The cell number of KM12L4 (A) and HT29 (B) parental and OxR cells were compared by MTT assay. At 24, 48, and 72 hours, both the KM12L4 and HT29 OxR cells exhibited reduced cell number compared with the respective parental cells.
Tables
- Table 1.
Cell cycle analysis of KM12L4 and HT29 OxR cells
48 hours Sub-G0 (%) G1 (%) S (%) G2 (%) KM12L4 parental 11 49 38 13 KM12 OxR 8 37 29 34 HT29 parental 3 68 24 8 HT29 OxR 3 53 27 20
Volume 12, Issue 14
