Depletion of CIP2A downregulates c-Myc protein expression in colon cancer cells.

<p>(<b>A</b>) Immunoblot analysis of CIP2A and c-Myc protein expression in Caco2, HCT116 and SW620 cells transfected with siRNA targeting CIP2A or control siRNA. Cells were harvested 72 h after transfection (n = 3 for each cell line). (<b>B</b>) Real-time PCR analysis of <i>CIP2A</i> and <i>c-Myc</i> mRNA expression in HCT116 cells transfected with siRNA targeting CIP2A or control siRNA (n = 3). (<b>C</b>) Depletion of CIP2A does not change activation status of AKT or its downstream targets. The panels show immunoblots of the indicated proteins and phosphorylated proteins (p) in Caco2, HCT116 and SW620 cells transfected with siRNA targeting CIP2A or control siRNA as before (n = 3 for each cell line).</p

CIP2A expression is regulated by MAPK signalling.

<p>Caco2, HCT116 and SW620 cells were treated with DMSO or the MEK inhibitor UO126 for 24(n = 3 for each cell line). (<b>A</b>) Immunoblot analysis of CIP2A and p-ERK protein expression in Caco2, HCT116 and SW620. (<b>B</b>) Real-time PCR analysis of <i>CIP2A</i> mRNA expression (*<0.05; **<0.005).</p

CIP2A is required for growth of HCT116 cells.

<p>(<b>A</b>) Immunoblot analysis of CIP2A and c-Myc protein expression in HCT116 cells infected lentiviral with shRNA targeting CIP2A or a ctr. shRNA. Numbers below lines indicate the c-myc protein expression relative to c-myc levels in control cells (n = 2). (<b>B</b>) Colony formation of HCT116 cells after 7 days. (<i>Top</i>), density of colonies stained with crystal violet; (<i>bottom</i>), representative of the indicated cultures (n = 3).</p

<i>CIP2A</i> mRNA expression is significantly correlated with advanced tumour stage.

<p>The panels show box and whisker plots documenting relative <i>CIP2A</i> mRNA levels in tumors stratified according to UICC stage (A) (UICC I vs. II p<.0001; II vs. III p = .0046; III vs. IV p = .0002), according to lymph node metastasis (B; N- vs. N+ p<.0001), according to distant metastasis (C; M0 vs. M1 p<.0001), and according to histological grading (D: G2 vs. G3 p = .0084).</p

CIP2A protein levels in colon cancer correlate with <i>CIP2A</i> mRNA expression.

<p>The panels show representative examples of immunofluorescence staining, showing CIP2A protein expression in cancer cells of patients with low (A+B) or high <i>CIP2A</i> (C+D) mRNA expression (A+C x100, B+D x200 magnification).</p

Patients with <i>CIP2A</i> high mRNA expression have an overall lower survival rate than patients with <i>CIP2A</i> low mRNA expression.

<p>The graphs show Kaplan–Meier curves of OS according to <i>CIP2A</i> mRNA expression. (red: <i>CIP2A</i> mRNA expression below median fold expression value of 10,5 above normal tissue), green: <i>CIP2A</i> mRNA expression above median fold expression value of 10,5 above normal tissue) (<b>A & B</b>) All patients with respect to <i>CIP2A</i> mRNA expression normalized to housekeeping gene (n = 75) (A: b2MG; B: GAPDH) (<b>C</b>) Patients in Stage UICC II with respect to <i>CIP2A</i> mRNA expression normalized to housekeeping gene GAPDH (n = 29) (<b>D</b>) Patients in Stage UICC III with respect to <i>CIP2A</i> mRNA expression normalized to housekeeping gene GAPDH (n = 21).</p

Additional file 4: Table S3. of Tumour stage distribution and survival of malignant melanoma in Germany 2002–2011

Relative 5-year survival of malignant melanoma patients diagnosed between 2002 and 2011, overall (UICC 0-IV, X) (N = 60 672) and for patients with invasive tumours (UICC I – IV, X) stratified by age, sex, UICC stage, ‘diagnosis during screening’ and place of residence (N = 49 351) (DOCX 39 kb

Additional file 3: Table S2. of Tumour stage distribution and survival of malignant melanoma in Germany 2002–2011

Malignant melanoma patients aged 35 years and above by age at diagnosis, sex, UICC stage, year of diagnosis, place of residence and ‘diagnosis during screening’, N = 34 739 (UICC 0 and X excluded) (DOCX 40 kb