Clinicopathological Characteristics And Survival Analysis For Gastroenteropancreatic Neuroendocrine Neoplasms & Micro Rna Expression Profile In Gastric Neuroendocrine Carcinoma

PartⅠ A retrospective analysis of clinicopathological characteristics and prognostic factors for gastroenteropancreatic neuroendocrine neoplasmsObjective: To investigate the clinicopathological characteristics of gastroenteropancreatic neuroendocrine neoplasms and its relationship with survival, in order to offer a theoretical reference for the clinical practice.Methods: Medical records and follows up of 93 GEP-NENs patients in the Sixth Affiliated Hospital of Shanghai Jiaotong University from May, 2003 to May, 2013 were collected and analyzed. This study focused on clinicopathological features. The primary endpoint was OS, and we ploted the survival curves of patients by Kaplan-Meier method. Log-rank method was used to examine the factors influencing prognosis.Results:1. Of all patients, male-to-female ratio was 1.4:1, the mean age was 57.1ys and median age was 53 ys. Most patients were 50 to 59 ys.2. Primary sites were the colorectum(33.3%), pancreas(25.8%), stomach(18.3%), appendix(12.9%) and intestine(9.7%). Most patients presented non-functional tumors(84.9%) with non-specific symptoms such as abdominal pain(63.4%) and gastrointestinal bleeding(28.0%). The manifestations of endoscopy and radiographicimage were tumor occupying lesion.3. GEP-NENs patients were diagnosed with neuroendocrine tumor(NET)(73.8%), neuroendocrine carcinoma(NEC)(23.8%) or mixed adenoneuroendocrine carcinoma(MANEC)(2.4%). The grading was G1(52.4%), G2(21.4%) or G3(26.2%).4. Positive rates of Cg A and Syn byimmuno-histochemistry examination were 89.5% and 70.9%, respectively, and there was significant difference(P = 0.002).5. At diagnosis, muscularis or serosa infiltration was found in 52.5% GEP-NENs patients, loco-lymphatic metastasis was in 20.0% cases and distant metastasis lied in 14.0% cases.6. A total of 91 patients(97.8%) underwent surgery. 17 patients were submitted forchemotherapy after operation.7. The mean survival was(92.7±6.4)months. The 1-, 3- and 5-year survival rates were 89.5%, 74.3% and 65.8%respectively.8. The prognosis of GEP-NEN is correlated with the pathological classification, grading, infiltrating extent, local lymphatic metastasis and condition of distant metastasis(P=0.003,P=0.001,P=0.038,P=0.015,P=0.006, respectively).Conclusions: Most GEP-NENs patients presented non-specific symptoms. Final diagnosis is base on pathological detection. The prognosis of GEP-NENs is more favorable than that of the adenocarcinomas of the digestive system. The prognosis of this tumor is correlated with the pathological classification, grading, infiltrating extent and condition of local lymphatic metastasis and distant metastasis. NEC/MANEC, G3, deep infiltrating, local lymphatic metastasis or distant metastasis predicts poor prognosis.Part Ⅱ Micro RNA expression profile in gastric neuroendocrine carcinomaObjective: To screening the different mi RNA expression in gastric neuroendocrine carcinoma and analyze the tumor related genes and signal pathway to lay a molecular basis for further study of mi RNA regulating mechanisms and molecular diagnostic tools of gastric neuroendocrine carcinoma.Methods: 9 gastric neuroendocrine carcinoma FFPE tissues and 9 para-cancer benign FFPE tissues were accumulated and total RNAs were isolated routinely. Expressions of mi RNAs in 3 gastric neuroendocrine carcinoma FFPE tissues and 3 para-cancer benign FFPE tissues were detected using micro RNA microarray. Target genes of differentially expressed mi RNAs were predicted with 3 databases. GO analysis and KEGG analysis were applied to determine the function of these target genes. Furthermore, some of the differentially expressed mi RNAs were chosen to be identified by quantitative real-time RT-PCR analysis.Results:1. The total RNAs extracted from FFPE tissue of gastric neuroendocrine carcinoma tissues and para-cancer benign tissues were suitable for further study.2. From the microarray results we found that a set of aberrant mi RNAs could clearly differentiate gastric neuroendocrine carcinoma tissues from para-cancer benign tissues. In particular, a significant increase of 17 mi RNAs and decrease of 21 mi RNAs were detected in gastric neuroendocrine carcinoma tissues in comparison with normal tissues(fold change≥2.0 and P v< 0.05). We found various kinds of target genes and pathway related with aberrant mi RNAs through bio-information analysis.3. Then the quantitative real-time RT-PCR analysis results showed that hsa-mi R-196a-5p, hsa-mi R-183-5p and hsa-mi R-18b-5p were significantly up-regulated and hsa-mi R-122-5p, hsa-mi R-378a-3p, hsa-mi R-422 a and hsa-mi R-490-3p were down-regulated in gastric neuroendocrine carcinoma tissues compared with normal controls, which in accordance with the microarray results.4. The aberrant mi RNAs, which might participate in either proliferation or migration by regulating target m RNA, are supposed to be potential biomarkers of gastric neuroendocrine carcinoma.Conclusions: It’s practicable to use gastric neuroendocrine carcinoma FFPE tissues for mi RNA expression analysis by microarray. It would be a feasible method in the research of GEP-NENs. This set of aberrant mi RNAs might be involved in gastric neuroendocrine carcinoma tumorigenesis and development. These results could expand our knowledge of molecular alteration involving in gastric neuroendocrine carcinoma pathogenesis and of the role of mi RNAs in human gastric neuroendocrine carcinoma. Whether the differentially expressed mi RNAs might become targets in human gastric neuroendocrine carcinoma deserve further research.