Journal: Translational Oncology

Article Title: Minichromosome Maintenance Expression Defines Slow-Growing Gastroenteropancreatic Neuroendocrine Neoplasms 1

doi: 10.1016/j.tranon.2016.07.006

Figure Lengend Snippet: Protein expression of MCM2 and MCM3 in small intestinal mucosa and SI-NENs (primaries and metastases). Immunohistochemistry (A): MCM2 and MCM3 were identified in crypts associated with the proliferative zone (PZ) (inset) which is the site of cells involved in renewing mucosa where the highest mitosis rates are expected. Additional cells were MCM3 positive at the base of crypts; these cells were not stained by MCM2 (red arrows). Dual staining with chromogranin A identified only a few (<2%) normal neuroendocrine cells to be MCM2 or MCM3 positive (yellow arrows). Both primary tumor and metastasis exhibited nuclear expression of MCM2 and MCM3, and expression of both proteins tended to be higher in metastasis in comparison to the primary tumor. This overexpression was also identified by Western blot (B), especially for MCM3, but this was not statistically significant (Kruskal-Wallis, MCM3: P = .06, MCM2: P = .15). DAPI: nuclei (blue), FITC: target marker (green), Cy5: chromogranin A (red-white arrows), dual-stained cells (yellow). PZ = proliferative zone. Mean ± SD.

Article Snippet: CNV was measured using PCR-Taqman Copy Number Reference Assay from Life Technologies according to the manufacturer's instructions using the primer sets Hs00575269_cn and Hs02422238_cn (MCM2) and Hs00378664_cn (MCM3).

Techniques: Expressing, Immunohistochemistry, Staining, Comparison, Over Expression, Western Blot, Marker

Journal: Translational Oncology

Article Title: Minichromosome Maintenance Expression Defines Slow-Growing Gastroenteropancreatic Neuroendocrine Neoplasms 1

doi: 10.1016/j.tranon.2016.07.006

Figure Lengend Snippet: Clinical relevance of MCM and Ki67 expression in SI-NENs. (A) MCM expression in normal mucosa tended to be higher than in neoplastic small intestinal tumors, whereas Ki67 was not significantly different between any tissue type (* P < .05 versus normal mucosa). (B) Ki67 and MCM2 expression was significantly increased (* P < .05) in grade 2 compared to grade 1 tumors. MCM3 was also increased, but this did not achieve statistical significance. (C) Correlation between MCM mRNA expression and Ki67 staining identified a relationship for MCM2 ( r 2 = 0.44, P < .05) but not for MCM3. Expression of either MCM significantly correlated with Ki67 mRNA expression (MCM2: r = 0.6, P = .0027, MCM3: r = 0.53, P = .0092) and with each other ( r = 0.69, P = .004). (D) High copy number variance was noted for all three markers in both primary tumors and in metastases; this was not seen in normal mucosa. Mean ± SD.

Article Snippet: CNV was measured using PCR-Taqman Copy Number Reference Assay from Life Technologies according to the manufacturer's instructions using the primer sets Hs00575269_cn and Hs02422238_cn (MCM2) and Hs00378664_cn (MCM3).

Techniques: Expressing, Staining

Journal: Translational Oncology

Article Title: Minichromosome Maintenance Expression Defines Slow-Growing Gastroenteropancreatic Neuroendocrine Neoplasms 1

doi: 10.1016/j.tranon.2016.07.006

Figure Lengend Snippet: Survival curves for AQUA analysis of MCM2 and MCM3 in SI-NENs. Low expressions of MCM2 (A) and MCM3 (B) were both associated with notably longer OS. Using a combination of proliferative markers, patients with low–proliferation score tumors (0-1; n = 21) had prolonged survival (median OS 12 years) compared to those with high–proliferation scores tumors (2-3; n = 24) (median OS years 6.5) (C, P = .06, HR 0.46). A high MCM3 expression was correlated with worst prognosis, whereas after more than 25-years of follow-up, the median OS of patients with low MCM3 expression was not reached in the subset of tumors that were only Ki67% positive (D, P = .019, HR 0.13).

Article Snippet: CNV was measured using PCR-Taqman Copy Number Reference Assay from Life Technologies according to the manufacturer's instructions using the primer sets Hs00575269_cn and Hs02422238_cn (MCM2) and Hs00378664_cn (MCM3).

Techniques: Expressing

Journal: Translational Oncology

Article Title: Minichromosome Maintenance Expression Defines Slow-Growing Gastroenteropancreatic Neuroendocrine Neoplasms 1

doi: 10.1016/j.tranon.2016.07.006

Figure Lengend Snippet: mRNA and protein expression of MCM2 and MCM3 in normal pancreas and pancreatic NENs (primaries and metastases). (A) MCM2 and MCM3 mRNAs were elevated in metastases in comparison to normal pancreas and to primaries, whereas Ki67 was elevated in metastases compared to normal pancreas (Kruskal-Wallis P < 0.0001, * P < .05 versus normal, # P < .05 versus primaries). (B) Ki67 and MCM mRNA expression as a function of grade identified no significant differences in pancreatic NENs. (C) Using Western blot, an elevation of MCM protein expression in pNEN metastases was confirmed (Kruskal-Wallis, MCM2: P = .24, MCM3: P = .44). Mean ± SD.

Article Snippet: CNV was measured using PCR-Taqman Copy Number Reference Assay from Life Technologies according to the manufacturer's instructions using the primer sets Hs00575269_cn and Hs02422238_cn (MCM2) and Hs00378664_cn (MCM3).

Techniques: Expressing, Comparison, Western Blot