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Molecular Cytogenetic Alterations of Human Hepatocellular Carcinoma Cell Models with Different Metastatic Potentials

Author: YangJiong
Tutor: TangZuoZuo
School: Fudan University
Course: Surgery
Keywords: Hepatocellular carcinoma Shift Relapse Cell lines G-banding Fluorescence in situ hybridization Comparative genomic hybridization Multiplex fluorescence in situ hybridization
CLC: R735.7
Type: PhD thesis
Year: 2003
Downloads: 196
Quote: 2
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Hepatocellular carcinoma is one of the most common tumors become a major obstacle to improve the prognosis of patients with liver cancer metastasis and recurrence. In order to explore exactly what cell accumulation of genetic alterations lead to transfer the formation of the malignant phenotype, Liver Cancer Institute, Fudan University on the basis of established human hepatocellular carcinoma metastasis model (LCI-D20), successfully completed the world's first high metastatic potential HCC cell lines MHCC97, and successful separation of cell clones with different metastatic potential MHCC97-H and MHCC97-L; these two cells were inoculated subcutaneously with amplified tumor formation, the incidence of lung metastasis after orthotopic liver transplantation were 100% and 40%, respectively. Recently built with higher metastatic potential the cell lines HCCLM3, occurred after the cells were inoculated subcutaneously with 100% of lung metastases. These cell lines and clones to study liver cancer metastasis-related cytogenetic abnormalities and molecular marker tools. My previous use of comparative genomic hybridization technology (CGH) found the 8p deletion and metastasis of HCC related liver cancer clinical specimens and the corresponding metastases in; CGH analysis of high-and low-transfer model LCI-D20 and LCI-D35 and MHCC97-H cell system of further confirmed this finding. On this basis, the study of 22 patients with clinical specimens heterozygous deletion (loss of heterozygosity, LOH) will further narrow the deletion region 8p23.3 and 8p11.2. To identify newly created with the same genetic background but there are significant differences in the metastatic potential of HCC cells also 8p deletion and figure out what they are in the other chromosome segments on the DNA level how similarities and differences, similarities and differences, but also has a kind of meaning, this project combined with G-banding technique, comparative genomic hybridization (CGH), multiplex fluorescence in situ hybridization (M-FISH) and sites or arm-specific fluorescence in situ hybridization, cell and molecular genetic studies of the cells, in order to understand the molecular cytogenetic anomaly identify metastasis-related genetic markers, to explore liver cancer transfer mechanisms to provide material. The establishment of the first part of fluorescence in situ hybridization-based molecular cytogenetic technology platform and its initial use of fluorescence in situ hybridization (Fluorescence in situ hybridization, FISH) and its associated molecular cytogenetic technology, the only few laboratories do is ideal. This section is intended FISH-based molecular genetics technology platform that I created. First high metastatic potential cells the clone MHCC97-H two-color fluorescence in situ hybridization FISH analysis of the indirect method. The use of commercially available 8 centromeric probes and their synthesis from the 8q23.1 region with different metastatic potential of human hepatoma cell model of the molecular cytogenetics Dr. abstracts BAC probe FISH hybridization, the results of two probes penetrating strength, specificity, and the detection are good. Preliminary found MHCC97 H clone chromosome 8 was non-interactive translocation trisomy change and its long arm signal to the region of the short arm of chromosome translocation and group B (chromosome 4). Pursuant to which the results of the use of purchased chromosome 8 specific painting probes (the Whole chromosomepainting WCP), clone hybridized the MHCC97 H, not only to optimize the direct method to probe the technical points, but also verify-color FlsH to the discovery. On the basis of the two-color FISH, we have established a comparative genomic hybridization (Co which arative genomichybridization, CGH) technology, and low metastatic potential or transfer of human hepatoma cell line SMMC-7721 were analyzed. CGH the technical indicators such as the target chip length of chromosomes, mitotic index as well as signal strength and homogeneity have reached the default settings. Detected in SMMC-7721 cell lines amplified region the IP31, lq25, 3p22 a pter SP, 6p21.3 a pter, 7p13 Pter, 8P23, SPll a q12, 9q22 qter the 10P12 Q21, 11pter a qZI llplZ a the QL4 14ql3 qter 15qls qter, 17p, Xp14 pter; deleted region 4q31 35 the of 3,1 3qZI 31,18 q gPZI, YqO Finally, we explore the multiplex fluorescence in situ hybridization (Multiplex fluoreseenee in situhybridization, M a FISH) technology, and HCCLM3 cell lines analyzed. The results show that the mixed specificity of the probe and brightness can be a sign chromosome translocation der (Y) t (Y; 18), der (3) t (3:20), der (4) (4; 8), der (10) t (9; 13), der (14) t (14:22), der (15) t (15:21), in addition to observation of the X chromosomes 1 and 8, the internal of the structural abnormalities. Two-color FISH chromosome painting the establishment of the the FISH, indirect CGH and MFISH technology, provided the conditions for the use of these techniques for different metastatic potential cell lines and clinical samples for analysis and comparison of molecular genetics. The second part of the metastatic potential of human hepatoma cell clones molecular cytogenetic analysis we combined with G-banding technique the CGH, MFISH, and chromosome arm and / or region-specific fluorescence in situ hybridization (FISH), from MHCC97 separation a different ability to transfer the cell clones MHCC97 a H, L MHCC97 a and HCCLM3 and molecular cytogenetic studies. Found: 1 .1 (X) (qlo), der (Y) t (y; 18) (qlZ; Pll), der (3) t (3:20) (P25: ql3), der (4) t ( 4; 8) (q31: q22), i (8) (QLO), der (14) t (14:22), India 13: ql3) chromosome abnormality in the three cell lines appear its flag chromosome, confirmed their common origin. 2.CGH three have their own specific genetic change characteristics, such as gqlZ 21,17 Q24 a qter, 18q22-23, a 13ql3, 13q22-31 appears only in in MHCC97 H; ZQ, LLQ, Yin different metastatic potential of human hepatoma cell model molecular cytogenetics research dissertation summary the 9q33 one ter only in an L MHCC97; In a 24 3qZI, 3q13.3 qter 9p12 a qter lopls, the 12p, 17plZ a QLZ 18pll .3 a pter, 19p, 2p23 a pter, etc. unique change in HCCLM3. The 8p23 deletion three cell there are no missing low transfer or transfer SMMC-7721 cells 8P area; In addition, we use the modified CGH method found in highly metastatic MHCC97 H cells?

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