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Chid L.,Center Hospital of Xian City
Cancer Research and Clinic | Year: 2015

Objective: To identify the differentially-expressed proteins of human gastric adenocarcinoma cell in primary or metastatic lymph node tissues by comparative proteomics technology, and to screen the specific metastatie-associated proteins so as to investigate the metastatic molecular mechanism of lymph node metastasis in gastric cancer. Methods:11 differential proteins were acquired previously from primary and metastatic lymphnode tissues in gastric adenocarcinoma patients by 2D-DIGE. Some selected differential protein spots were identified by PMF based on MALDI-TOF-MS and database search. Immunohistoehemieal staining of HSP70 was used to evaluate the reliability of the proteomic analysis results. Results: After analyzed on 11 differential proteins by in-gel trypsin digestion and MALDI-TOF-MS-based PMF analysis, a total of 5 differential proteins were identified by searching Mascot-database, HSP70' s 8 isoform 2 variant, chaperonin, chaperonin, leucine aminopeptidase, predicted: hypothetical protein XP-515584. Among the differential proteins identified, the levels of HSP70, chaperonin, leucine aminopeptidase expression had a significant up-regulation in gastric primary cancer compared with metastatic lymph node. HSP70 expression rate increased with the metastasis of lymph node and the progress of gastric cancer, agreed with the proteomics results. Conclusions: They are similar in differentially-expressed proteins in primary or metastatic lymph node tissues because of the uniformity in source and differention. There are few protein changes in cancer cells between them, taking part in the metastatic of gastric cancer. HSP70 takes part in the progress of gastric cancer and relates to the metastasis of lymph node and malignant degree. Source


Lili C.,Center Hospital of Xian City | Guoqing Y.,Center Hospital of Xian City | Haiwei W.,Center Hospital of Xian City | Yan C.,Center Hospital of Xian City
Cancer Research and Clinic | Year: 2015

Objective To investigate the human papillomavirus 16/18 (HPV16/18) infective status and its relationship with the expression of pl6, CK17, Ki-67 in immature squamous metaplasia(IM), cervical intraepithelial neoplasia (CIN) and cervical carcinomas (CC). Methods In situ hybridization (ISH) and immunohistochemistry were applied to detect the expression of HPV16/18, pl6, CK17, Ki-67 on tissues from 30 IM, 60 CIN and 30 CC. Results The expression rates of HPV16/18 in LSIL, HSIL and CC were 30 % (9/30), 60 % (18/30) and 77 % (23/30), higher than that in IM [6.7 % (2/30)]. HPV16/18 was positively expressed with the progression of cervical squamous epithelial lesions. The expression rates of CK17 in IM was 90 % (27/30), higher than that in LSIL, HSIL and CC [17 % (5/30), 10 % (3/30), 6.7 % (2/30)]. The expression rates of pl6, Ki-67 were 83 % (25/30) and 87 % (26/30) in LSIL, 90 % (27/30) and 93 % (29/30) in HSIL, 97 % (29/30) and 97 % (29/30) in CC, higher than that in IM [13 % (4/30) and 10 % (3/30)]. The expression of p16, Ki-67 were particularly seen in HPV16/18-positive HSIL and CC, and the correlation were observed. Conclusion HPV16/18 infection is highly associated with the degree of squamous intraepithelial neoplasia and carcinomas of cervix and upregulated pl6 and Ki-67, which suggested that HPV16/18 maybe cause mutation of pl6 gene. With the progression of cervical squamous epithelial lesions, positive expression rates of CK17 were decreased compared with increased of HPV16/18, p16, Ki-67. Combined detection of HPV16/18, pl6, Ki-67 is helpful for diagnosis and classification of cervical squamous epithelial lesions. Source


Chai L.,Center Hospital of Xian City | Chen Y.,Center Hospital of Xian City | Wang H.,Center Hospital of Xian City | Yang G.,Center Hospital of Xian City | Zheng C.,Center Hospital of Xian City
Cancer Research and Clinic | Year: 2015

Objective To establish differential protein expressing profiles of human gastric adenocarcinoma cell in primary or metastatic lymph node tissues by two-dimensional differential gel electrophoresis (2-D DIGE) so as to investigate the metastatic molecular mechanism of the gastric cancer. Methods After obtaining 6 samples of human primary gastric cancer and metastatic lymph node tissues, with manual no-staining frozen sections microdissection, human gastric adenocarcinoma cells from primary or metastatic lymph node tissues were isolated, and then the total proteins were extracted and purified. Highly sensitive 2-D DIGE was used to separate the total protein differentially expressed in the cells. The proteins were visualized by using a fluorescence scanner at appropriate wavelengths for Cy2, Cy3 and Cy5 dyes (Typhoon 9400). Image analysis was carried out with the DeCyder-Differential analysis software (Biological Variation Analysis version 5.0). Results Not only can the study procure defined adenocarcinoma cell populations from gastric primary or metastatic lymph node tissues, but also can resolve the problem of the change in 2-D DIGE patterns because of the varying in protein changes owing to dyeing. All these showed that the technique was simple, easy to perform, versatile and of particular usefulness when laser capture microdissection (LCM) was practically unavailable. The 2-D DIGE patterns with high resolution and reproducibility from adenocarcinoma cells in gastric primary or metastatic lymph node tissues were obtained. The number of spots in Gell, Gel2 were 1 416 (similar 1 062, decrease 277, increase 77), 1 299 (similar 1 050, decrease 157, increase 92), respectively. A total of 11 differential proteins were acquired by image analysis with DeCyder-Differential analysis software (Biological Variation Analysis version 5.0). Conclusions In this report, a simple, easy to proform method of protein epuration, manual no-staining frozen sections microdissection is described, and have used the highly sensitive 2-D DIGE for the identification of proteins differentially expressing in human gastric adenocarcinoma cells from primary or metastatic lymph node tissues. These results provide a fundamental basis for further study of metastatic mechanism of gastric cancer and screen its specific markers. Source

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