Proteomics of differentially-expressed proteins between human primary and lymph node metastatic gastric adenocarcinoma / 肿瘤研究与临床

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 metastatic-associated proteins so as to investigate the metastatic molecular mechanism of lymph node metastasis in gastric cancer. Methods 11differential 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. Immunohistochemical 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, HSP70ˊs 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.

Preliminary differential proteome analysis of the human primary or lymph node metastatic gastric adenocarcinoma by two-dimensional differential gel electrophorosis / 肿瘤研究与临床

Objective To establish differential protein expressing profiles of human gastric adenocarcinoma cell in primary or metastatic lymph node tissues by two-dimensional differential gel electrophorosis (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 Gel1,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.

Cytotoxicity of a new biomedical titanium alloy Ti-25Nb-10Ta-1Zr-0.2Fe / 中南大学学报(医学版)

OBJECTIVE@#To evaluate the cytotoxicity of a new type of titanium alloy Ti-25Nb-10Ta-1Zr-0.2Fe by studying the induced proliferation of L929 cells in contrast with other titania widely used in clinical practice.@*METHODS@#The cell line was treated with extracting liquid containing different concentrations of titanium alloys. The number and morphology of cells was observed under an inverted phase contrast microscope. MTT was used to measure the relative growth rate (RGR) and judge the cytotoxicity grade. Flow cytometry was used to observe cell cycle progression.@*RESULTS@#The RGR of TNTZ group cells at the 3 time points was (93.7±0.8), (100.6±0.4), and (106.4±0.3); the cytotoxicity grade was 1, 0 and 0 after treating for 1, 3 and 5 days; with influence on neither the cell morphology nor the cell cycle. The flow cytometry showed that the sequence of S phase cells was Ti>TNTZ>TC4>blank control >TC4ELI, with no significant difference (P>0.05). None of the 4 materials inhibited the cell proliferation.@*CONCLUSION@#The cell morphology and proliferation are not affected by TNTZ. The new titaniu alloys shows good cyto-compatibility. The cytotoxicity is grade 0, meeting the clinical application standard.

Study on the expression of PTEN and P53 in human primary gallbladder carcinoma(HGC) / 中国癌症杂志

Background and purpose:To study the expression of PTEN and P53 and analyse their roles in HGC oncogenesis,and to discover a good marker for diagnosis and prognosis of HGC.Methods:The expression of PTEN mRNA,PTEN protein and P53 protein were detected by in situ hybridization(DNA-RNA) and immunohistochemistry(SP method) in 64 cases of HGC.We took 22 cases of gallbladder adenoma and 10 cases of chronic cholecystitis as controls.Results:① The positive expression rates of PTEN mRNA and protein in HGC(56.3%,59.4%) were significantly lower than that in gallbladder adenoma(81.8%,90.8%) and chronic cholecystitis(100%,100%)(P