Daniel Seidl

Characterization of Spontaneous and TGF-β-Induced Cell Motility of Primary Human Normal and Neoplastic Mammary Cells In Vitro Using Novel Real-Time Technology

The clinical complications derived from metastatic disease are responsible for the majority of all breast cancer related deaths. Since cell migration and invasion are a prerequisite for metastasis their assessment in patient cancer cells in vitro may have prognostic value for the tumors metastatic capacity. We employed real-time cell analysis (RTCA) on the xCELLigence DP system to determine in vitro motility of patient-derived primary human breast cancer epithelial cells (HBCEC). Initially, the RTCA assay was validated using established human breast cancer cell lines with either an invasive (MDA-MB-231, MDA-MB-435s) or a non-invasive phenotype (MCF-7, MDA-MB-468), and primary NSCLC cells (Tu459). Previous standard assays of cell migration/invasion revealed that only MDA-MB-231, −435s, and Tu459 cells exhibited spontaneous and TGF-β1-stimulated migration and invasion through a Matrigel barrier. In the present study, the TGF-β1-stimulated activities could be blocked by SB431542, a potent kinase inhibitor of the TGF-β type I receptor ALK5. Application of the RTCA assay to patient-derived tumor cells showed that 4/4 primary HBCEC and primary NSCLC cells, but not normal human mammary epithelial cells (HMEC), displayed high spontaneous migratory and invasive activity which correlated with higher MMP-2 expression and uPA protein levels in HBCEC compared to HMEC. Upon treatment with TGF-β1, HBCEC exhibited morphologic and gene regulatory alterations indicative of epithelial-to-mesenchymal transition. However, exclusively the invasive but not the migratory activity of HBCEC was further enhanced by TGF-β1. This indicates the requirement for molecular, e.g. integrin interactions with Matrigel components in HBCEC in order to become responsive to pro-invasive TGF-β effects. Together, these results show for the first time that tumorigenic HBCEC but not normal HMEC possess a strong basal migratory as well as a basal and TGF-β1-inducible invasive potential. These findings qualify the RTCA assay as an in vitro migration/invasion testing system for patient-specific primary breast cancer cells.

Do we need 5-FU in addition to cisplatin for chemoradiation of locally advanced head-and-neck cancer?

OBJECTIVES To compare chemoradiation with cisplatin alone or cisplatin plus 5-FU for locally advanced squamous cell carcinoma of the head-and-neck (SCCHN). MATERIALS AND METHODS The outcomes of 142 patients who received chemoradiation with cisplatin alone for locally advanced SCCHN were retrospectively compared to 170 patients who received cisplatin plus 5-fluorouracil (5-FU). The outcomes compared included loco-regional control (LRC), metastases-free survival (MFS), overall survival (OS) and adverse events. RESULTS Although patients who received cisplatin alone had a significantly worse performance status, 81% of these patients completed planned chemotherapy compared to 73% of patients in the cisplatin plus 5-FU group (p=0.18). Radiotherapy breaks >1week were necessary in 14% and 23% of patients, respectively (p=0.09). The 5-year LRC rates were 69% after cisplatin alone and 68% after cisplatin plus 5-FU (p=0.71). The 5-year MFS rates were 72% and 62%, respectively (p=0.37), and 5-year OS rates were 60% and 45%, respectively (p=0.066). On multivariate analysis, cisplatin alone was significantly associated with improved OS (RR 1.35; 95%-CI 1.09-1.69; p=0.006). Nausea/vomiting, pneumonia/sepsis and late adverse events occurred more common in the cisplatin plus 5-FU group. CONCLUSION Given the limitations of a retrospective study, chemoradiation with cisplatin alone appeared associated with fewer adverse events and better OS than with cisplatin plus 5-FU in patients with locally advanced SCCHN. Thus, 5-FU in addition to cisplatin may be omitted for these patients. A randomized trial is warranted to confirm these findings.

Chemoradiation of locally advanced squamous cell carcinoma of the head-and-neck (LASCCHN): Is 20 mg/m2 cisplatin on five days every four weeks an alternative to 100 mg/m2 cisplatin every three weeks?

OBJECTIVES To compare chemoradiation with 100mg/m(2) cisplatin every three weeks to 20mg/m(2) on five days every four weeks for locally advanced squamous cell carcinoma of the head-and-neck (LASCCHN). MATERIALS AND METHODS In 230 patients receiving chemoradiation for LASCCHN, 100mg/m(2) cisplatin every three weeks (N=126) and 20mg/m(2) cisplatin on five days every four weeks (N=104) were retrospectively compared. Chemoradiation plus eleven characteristics (T-/N-classification, performance score, gender, age, tumor site, grading, surgery, radiation technique, pre-chemoradiation hemoglobin, cumulative cisplatin dose) were analyzed for locoregional control (LRC), metastases-free survival (MFS) and overall survival (OS). Chemoradiation groups were compared for adverse events. RESULTS On univariate analyses, chemoradiation had no impact on LRC (p=0.53), MFS (p=0.67) and OS (p=0.14). On multivariate analysis of LRC, T-classification (p=0.045) and hemoglobin (p<0.001) were significant. On multivariate analysis of MFS, performance score (p=0.028) was significant. On multivariate analysis of OS, performance score (p=0.009) and hemoglobin levels (p=0.002) achieved significance. Chemoradiation with 100mg/m(2) cisplatin was associated with more pneumonia/sepsis (p=0.003), grade ⩾2nausea/vomiting (p<0.001), grade ⩾2 nephrotoxicity (p=0.005), grade ⩾2 xerostomia (p=0.002), grade ⩾3 hematotoxicity (p=0.052) and grade ⩾2 ototoxicity (p=0.048). CONCLUDING STATEMENT 20mg/m(2) cisplatin on five days every four weeks was associated with fewer adverse events than 100mg/m(2) cisplatin every three weeks. 100mg/m(2) cisplatin was not significantly superior to 20mg/m(2) cisplatin regarding LRC, MFS and OS. Given the limitations of a retrospective study, 20mg/m(2) cisplatin appeared preferable. The results should be confirmed in a randomized trial.