Figure 1

Acute anticancer effect of MST-312 inversely correlates with telomere length of cancer cells. (A–C) In vivo anti-tumour effect of MST-312 in mouse xenograft models. Human breast cancer HBC-4 cells were subcutaneously injected into nude mice. Mice were treated with intratumoural (A), intravenous (B) or oral administration (C) of vehicle or MST-312. Error bar indicates standard deviation. Upper and lower graphs indicate the relative tumour volume and body weight (BW) of the mice, respectively. (D) In vitro anti-proliferative effect of MST-312 on the JFCR39 panel of 39 human cancer cell lines. Cells were treated with indicated concentrations (molar) of MST-312 for 48 h and then cell numbers were quantitated. (E) Fingerprint of MST-312 sensitivity. GI₅₀ values of MST-312 quantitated by (D) and the average of all cell lines was defined as zero. (F) Telomere blot analysis of JFCR39. Genomic DNA was prepared and subjected to Southern blot analysis with the [³²P]-labelled telomeric probe to detect telomeric restriction fragments (TRFs). Two different blots were derived from the same experiment and were processed in parallel. Their border was indicated by a dotted line. (G) Expression of telomere-related proteins in JFCR39. Cell lysates were prepared and subjected to western blot analyses with indicated primary antibodies. For each antibody blot and Coomassie stain, three different blots or gels were derived from the same experiment and were processed in parallel. Their borders were indicated by dotted lines. Each blot/gel contains NCI-H23 cells as a calibration standard. Full-length blots were presented in Supplementary Fig. S4. (H) Telomerase activity in JFCR39 cells. Cell lysates were prepared and subjected to TRAP assay. Average telomerase activity of all cell lines was defined as zero. (I) Two-dimensional hierarchical cluster analysis of the telomere-related bioparameters. The clustering result was generated by Cluster (ver. 3.0) and Java TreeView (Ver. 1.1.6r4). Orange dots: four of six shelterin components (TRF1, POT1, TIN2 and TPP1); yellow dots: telomerase components (hTERT and Dyskerin) and telomerase activity; pink dots: two shelterin components (TRF2, RAP1); light blue dots: MRN complex (MRE11, NBS1 and RAD50). (J) Correlation between the cell sensitivity to MST-312 and the length of the mean TRF length (left) or telomere signals quantitated by HPA assay (right). In these graphs, the super long telomere cells (NCI-H23, DMS114 and LOX-IMVI) were excluded from the calculation of correlation coefficient.