论文标题:G-CSF在人实体肿瘤中的表达及其促增殖作用的实验研究
The Experimental Study of G-CSF Expression and Promoting Cell Proliferation in Human Solid Carcinomas
论文作者 沙丹
论文导师 张灿珍；陈明清,论文学位 硕士,论文专业 肿瘤学
论文单位 昆明医学院,点击次数 117,论文页数 50页File Size2283k
2003-06-01论文网 http://www.lw23.com/lunwen_1406157/ 粒细胞集落刺激因子,实体肿瘤,表达,增殖
granulocyte colony-stimulating factor;solid carcinoma;expression;proliferation
目的 研究7种人实体肿瘤细胞株(GLC、YTMLC、BIU-87、T-24，U-251、Tca-8113和KB)体外培养上清液中G-CSF的分泌水平及外源性rhG-CSF对上述细胞增殖的影响，从而为临床应用rhG-CSF安全与否提供理论依据。方法 用人G-CSF ELISA试剂盒检测培养上清液中G-CSF的分泌水平；对分泌G-CSF的细胞，继续研究其不同时间点G-CSF的分泌水平。用血细胞计数板计数法、MTT法和集落形成实验检测不同浓度rhG-CSF(0ng／ml-1000ng／ml)对7种肿瘤细胞增殖的影响。结果 1、细胞株T-24分泌大量G-CSF(＞2000pg／ml)，其分泌量随培养时间延长而增加；其余6种肿瘤细胞培养上清中未检测到G-CSF分泌。2、随着rhG-CSF浓度的增加，KB细胞的细胞计数、OD值、增殖率和集落数有逐渐增多的趋势，且在rhG-CSF浓度为100ng／ml达最大值[分别为(10.6±0.1)×10~4个／毫升、2.343±0.105、35.2％和257±24.8个／孔]，与对照组相比，差异有显著性(P＜0.05)；KB细胞的其余rhG-CSF浓度组与对照组相比，差异无显著性(P＞0.05)。其它肿瘤细胞未观察到rhG-CSF的促增殖效应。3、细胞株GLC可在含5％小牛血清的培养基中存活和增殖。结论 1、细胞株T-24向培养上清分泌大量G-CSF，其分泌量随时间延长而增加；2、rhG-CSF对细胞株KB有促增殖作用，并在100ng／ml时达最大值，其促增殖的剂量依赖效应不明显；3、细胞株GLC可能通过分泌一定的生长因尾明医学晚硕士攀位论文子而在含5%小牛血清的培养基中存活和增殖;4、对G一CSF受体阳性的肿瘤患者，不宜在没有强有力放、化疗的配合下大剂量使用rhG一CSF。
Objective: In order to provide the experimental evidence for secure application of rhG-CSF, G-CSF expression and promoting cell proliferation in 7 kinds of human solid carcinoma cell lines were investigated. Methods: The expression of G-CSF from these cell lines was detected by Enzyme-Linked Immunoabsorbent Assay Kit. The G-CSF production of G-CSF positive cell line was further investigated at different time point. rhG-CSF promoting cell proliferation was studied by cell count, MTT and colony formation assay. Results: 1. Cell line T-24 produced large amount of G-CSF(>2000ng/ml), which increased with time going. At the same time, G-CSF expression was not observed in other 6 cell lines; 2. rhG-CSF promoted cell line KB proliferation in vitro. After rhG-CSF administration at a concentration of 100ng/ml, the maximum of cell count, OD value, proliferative rate and colony number of cell line KB reached (10.6±0.1) × 104/ml, 2.343 ± 0.105, 35.2% and 257±24.8/well respectively, which had significant difference compared with control group (P<0.05). No proliferative effect was observed in other 6 cell lines; 3. Cell line GLC surviving and proliferation was observed in medium containing 5% calf bovine serum. Conclusions: 1. Cell line T-24 produced large amount of G-CSF, which increased with time going; 2. Exogenous rhG-CSF promoted KB cellproliferation in vitro and reached its maximal proliferative effect at a concentration of 100ng/ml; 3. Cell line GLC can survive and proliferate in medium containing 5% calf bovine serum probably by secreting some growth factors; 4. For patients with G-CSF receptor positive carcinoma, high dose G-CSF shouldn"t be administrated without combined treatment of radiotherapy or chemotherapy.
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