Fig. 2: GCLM translocates to the nucleus up platinum drug treatment. | Nature Communications

Fig. 2: GCLM translocates to the nucleus up platinum drug treatment.

From: Nucleus-translocated GCLM promotes chemoresistance in colorectal cancer through a moonlighting function

Fig. 2

a IHC staining and scores of GCLM expression in primary CRC tumor (T) and adjacent normal tissues (N) (n = 406, CRC tissue specimens). Scale bar = 50 μm. The data are presented as a box-and-whisker graph (minimum–maximum), and the horizontal line across the box indicates the median (a right). b Overall survival and disease-free survival assays of CRC patients based on the GCLM protein level in (a). c IHC staining of GCLM expression in CRC patients receiving FOLFOX or XELOX chemotherapy. The blue arrowheads indicate the GCLM expression in nucleus. The correlation between GCLM expression and the response of patients to the standard chemotherapy (right), which is presented in the form of the percentage of total samples. (n = 58, CRC tissue specimens). Scale bar = 50 μm. d Cell viability of CRC cells overexpressing control or rGCLM WT, which were treated with PBS, oxaliplatin (40 μM for HCT116, 80 μM for DLD1, 24 h) or BSO (150 μM, 24 h). e IB detection of total, cytoplasmic and nuclear GCLM expression in HCT116 cells treated with 40 μM oxaliplatin (Oxa), cisplatin (CDDP) and carboplatin (CBP), and 20 μM irinotecan (CPT-11) and 5-fluorouracil (5-FU) for 24 h. f IF staining showing the localization of GCLM in HCT116 cells with the treatment of PBS, oxaliplatin or cisplatin (40 μM, 24 h). Scale bar = 10 μm. Cell viability (g) and apoptotic cells (h) among HCT116 cells overexpressing control, nuclear GCLM (NLS) or C193/194 A mutant (NLS-CA) with oxaliplatin or cisplatin treatment (40 μM, 24 h). i Photographs and tumor volumes, weights analysis of CDX model after the implantation of HCT116 cells overexpressing control or nuclear GCLM, followed by intraperitoneal injections of PBS or oxaliplatin (5 mg/kg). IB experiments were repeated three times and n = 3 biologically independent experiments in (d, g, h) and n = 5 mice in (i). The data are presented as the mean ± S.D (d, g, h, i). The P values were calculated by two-tailed paired Student’s t test (a), Kaplan–Meier analysis (log-rank test) (b), two-sided chi-square test (c), one-way ANOVA (h, i right), and two-way ANOVA (d, g, i middle). PD progressive disease, SD stable disease, PR partial response, CR complete response, NLS nuclear localization signals.

Back to article page