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Objective To establish the cell line from specimens of resectable human gastrointestinal stromal tumors (GIST) and to verify the characteristics of cell biology in vitro. Methods The tissues from biopsies of human GIST were cultured in RPMI 1640 media supplemented with 10% fetal bovine serum. After growing to 90% confluence,the cells were detached for subculture and their characteristics,including morphology,growth kinetics,karyotype analysis,immunohistochemical analysis and tumorigenicity in nude mice were determined. Results GIST named GIST-H1 was successfully established. The cell line was passaged for more than 60 times 1 year. The characteristics demonstrated: The population doubling time calculated in the log phase of growth was 47.5 h. The cloning efficiency in the soft agar averaged 24.8%.Electronmicroscopically,there were rich ribosomes and mitochondrion in the cytoplasm. Immunohistochemical analysis showed CD117(+),SMA(+),dog-1(+),CD34(-),and S-100(-).Karyotype analysis illustrated aneuploidy with the modal chromosomal number 60-98.The GIST cells transplanted in nude mice had high tumorigenicity. Conclusion The immortalized GIST cells are devoloped in vitro and have specific characteristics of GIST.
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Objective To study the expressions of heat shock protein 27(HSP27)in brain tissue and the changes of antibody of HSP27 in serum after transient cerebral ischemic reperfusion in gerbils.Methods A cerebral ischemia-reperfusion model in gerbils was established by clamping both common carotids.Sixty gerbils were randomly divided into normal control group,sham operation group and ischemia-reperfusion(IR)group.The expressions of HSP27 in brain tissue were detected by immunohistochemistry technique and the changes of antibody of HSP27 were detected by ELISA technique at the 6th hour,1st day,3rd and 7th day after IR.Experimental results were analyzed with the statistic soft package of Spss11.5.Results Experimental results revealed that HSP27 were expressed in neurons and gliacytes of the brain tissue in the 60 gerbils.There was mild expressions of HSP27 in neurons and gliacytes in normal control group and sham operation group, but it has no significant difference between the two groups(P＞0.05).Compared with normal control group,the expressions of HSP27 in neurong and gliacytes of ischemia-reperfusion gerbils were evidently increased at the 6th hour,1st day,3rd day and 7th day,especially at the 3rd day(P＜0.01).Conclusions Expressions of HSP27 in neurons and gliacytes of gerbils were increased markedly.This suggested that the increased expressions of HSP27 could protect the brain tissue from damage.The changes of antibody in serum of normal control group,sham operation group were evidently less than that in group IR,which suggest the HSP27 antibody may damage our body.
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AIM: Eight melanoma cell lines were establi sh ed from human melanoma specimen and designated as HME 1-HME 8, and their charact eristics of cell biology and immunology in vitro were studied. METHODS: The tissues from biopsies of human melanoma were cultur ed in RPMI 1640 media. After growing to 90% confluence, the cells were detached for subculture and then their characteristics, including morphology, growth kine tic, tumor antigen and tumorigencity in nude mice were studied. RESULTS: These cell lines have been passaged more than 100 times within 2 years. Their characteristics demonstrated: ① The population doubling time calculated in the log phase of growth were 34.2-59.5 h. ② The cloning ef ficiency in soft agar averaged 8.6%-26.2%. ③ The melanoma cells transplan t ed in nude mice were high tumorigencity. ④ Karyo-type analysis showed that aneu ploidy with the modal chromosomal number 64-117. ⑤ Rich ribosomes and melanoid grains in the cytoplasm were observed under electronic microscope. ⑥ Immunohist ochemistry analysis showed that there were a lot of brown grains in the cytoplas m. ⑦ Detection of tumor-antigen demonstrated that melan-A antigen was 75.0%, M AGE-1 was 50.0%, MAGE-2 was 37.5%, MAGE-3 was 62.5%, respectively, for eight melanoma cell lines. CONCLUSION: The melanoma cells have immortalized after being cul tured in vitro and has specific characteristics of malignant melanoma.