Figure 1.
Sensitivity of wild-type cell lines to paclitaxel treatment.
A: Microscopic assessment of apoptosis in Ishikawa and Hec1B wild-type cells after treatment with paclitaxel in the following dosages: 0 nM, 5 nM, 10 nM, 50 nM, 100 nM, 250 nM and 500 nM. Results are representative of 5 independent experiments. Standard errors of the mean are indicated. B: Cell metabolic activity assessed with a proliferation assay (MTS, Promega) in Ishikawa and Hec1B wild-type cells after treatment with paclitaxel in the following dosages: 0 nM, 1 nM, 10 nM, 50 nM, 100 nM, 250 nM and 500 nM. Results are representative of 3 (Ishikawa) and 2 (Hec1B) independent experiments.
Figure 2.
Ishikawa cell line experiments after stathmin knock-down.
A: Immunoblot after transfecting cells with a stathmin lentiviral shRNAmir (‘stmn kd’) or a non-silencing control (‘non sil’) as well as the parental cell line (wild-type; ‘WT’). Blots were stained for stathmin, and β-actin for loading control. B: Ishikawa wild-type cell line, non-silencing and stathmin knock-down after treatment with paclitaxel for 24 h in the following dosages: 0 nM, 5 nM, 10 nM, 50 nM, 100 nM, 250 nM and 500 nM. The level of fragmentation of the cells is indicated in an insert, as a proxy of progression in the apoptotic process. Diamonds; wild-type, triangles; non-silencing and crosses; stathmin knock-down cells. C: Immunoblot of Ishikawa wild-type, control (non-silencing) and stathmin knock-down cell lines after treatment with paclitaxel for 24 h in the following dosages: 0 nM, 100 nM and 250 nM. The blot was stained for cleaved PARP and stathmin, with β-actin serving as loading control. D: Left: Ishikawa wild-type (‘WT’) cell line and Right: Ishikawa stathmin knock-down (‘Stmn kd’) cell line. Microscopic images of cells after treatment for 24 h with 0 nM (top row) or 500 nM (bottom row) paclitaxel also demonstrating increased fragmentation rate for the stathmin knock-down Ishikawa cells (right lower panel) compared to wild-type (left lower panel).
Figure 3.
Hec1B cell line experiments after stathmin knock-down.
A: Immunoblot after transfecting cells with a stathmin lentiviral shRNAmir (‘stmn kd’) or a non-silencing control (‘non sil’) as well as the parental cell line (wild-type; ‘WT’). Blots were stained for stathmin, and β-actin for loading control. B: Hec1B wild-type cell line, non-silencing and stathmin knock-down lines, after treatment with paclitaxel for 24 h in the following dosages: 0 nM, 5 nM, 10 nM, 50 nM, 100 nM, 250 nM and 500 nM. C: Left: Hec1B wild-type (‘WT’) cell line and Right: Hec1B stathmin knock-down (‘Stmn kd’) cell line. Showing microscopic images of cells after treatment for 24 h with 0 nM (top row) or 500 nM (bottom row) paclitaxel demonstrating increased cell death for the stathmin knock-down Hec1B cells (right lower panel) compared to Hec1B wild-type (left lower panel).
Figure 4.
Stathmin protein expression in relation to clinical parameters.
A: Pictures representative of weak immunohistochemical stathmin staining (top) and strong (high or pathologic) stathmin staining (bottom) in endometrial carcinoma. Bars (right lower corner) measure 40 μm. B: Clinical response to paclitaxel for all endometrial carcinoma patients with evaluable disease according to RECIST criteria and separated for normal versus high stathmin level. Poor response (RECIST: static disease or disease progression) indicated in blue, good response (RECIST: complete or partial response) indicated in red. C: Comparison of high (pathologic) stathmin protein level in primary and metastatic lesions.
Table 1.
Characteristics of patients receiving paclitaxel or other treatment for metastatic endometrial cancer (n = 78).
Figure 5.
Disease specific survival after primary treatment for endometrial carcinoma patients (Kaplan-Meier curves) related to stathmin protein expression by IHC in primary tumor.
A: All patients with complete data (n = 476). Number of disease specific events between brackets. B: All patients with metastatic disease who received paclitaxel treatment (n = 38). Number of disease specific events between brackets. C: All patients with metastatic disease who received different treatments (n = 43). Number of disease specific events between brackets.