无功能垂体腺瘤来源细胞-F的核型分析

Karyotype analysis of non-functioning pituitary adenomas derived cell-F

目的研究体外培养的人无功能垂体腺瘤来源细胞-F的染色体与肿瘤遗传学、生物学行为之间的关系。方法原代培养人无功能垂体腺瘤细胞并传代,将第10、20、30代的腺瘤细胞以直接阻滞法获取染色体,结合DNA倍体分析和G显带法对染色体变异进行观察、分析;并采用电镜对细胞进行超微结构观察,免疫细胞化学检测蛋白,光化学法测定上清液激素表达。结果体外培养的人无功能垂体腺瘤细胞为嵌合体型细胞。正常染色体中1、3、7、10号缺失,4、6、18、19、22号有1条染色体,2、8、9、11、12、13、15、20、22号有2条染色体,5、16、17号有3条染色体,14号有4条染色体,X1条染色体有脆性位点。实验组第5、10、15、20、25、30代细胞上清液中催乳素分泌水平分别为:(2.97±0.08)ng/ml、(2.91±0.97)ng/ml、(0.92±0.30)ng/ml、(1.12±0.50)ng/ml、(0.62±0.17)ng/ml、(0.40±0.06)ng/ml,与对照组相比差异均有统计学意义(均P<0.05)。电镜发现:细胞表面微绒毛,线粒体灶性肿胀和空化,分泌颗粒分布稀疏,直径为150～220 nm。免疫细胞化学结果:人垂体催乳素检测阳性,而其他激素蛋白检测阴性。结论体外培养生长的人垂体无功能腺瘤细胞存在染色体畸变,形成同源嵌合体。这些畸变的染色体与垂体腺瘤的增殖、分泌功能相关。

Objective To study the relationships between chromosomes and both cancer genetics and biological behavior of in vitro cultured human non-functioning pituitary adenomas derived cell-F.Methods Primary human non-functioning pituitary adenomas derived cell-F was cultured and passaged in vitro.Cytokinesis-block assay was used in passages 10,20,30 to get chromosomes.DNA ploidy analysis and chromosome G-band analysis were applied for chromosomal alteration.The electron microscope,immunocytochemistry and photochemical method were also used to detect cell ultrastructure,protein expression and hormone content in the supernatant.Results In vitro cultured human non-functioning pituitary adenomas derived cell-F was a chimeric karyotype.The chromosome deletion involved No.1,3,7 and 10.There was 1 chromosome in No.4,6,18,19 and 22,2 chromosomes in No.2,8,9,11,12,13,15,20 and 22,3 chromosomes in No.5,16 and 17,4 chromosomes in No.14 and a fragile site on the X chromosome.The prolactin levels in the supernatants of the cells at passages 5,10,15,20,25,30 in the experiment group were 2.97±0.08,2.91±0.97,0.92±0.30,1.12±0.50,0.62±0.17,and 0.40±0.06 ng/ml respectively and had significant differences compared with control group（all P0.05）.Electron microscopy showed the microvilli on the cell surface,swollen and hollowed mitochondria and sparse secretory granules with the diameters ranged from 150 to 220 nm.The immunocytochemical assay showed that human prolactin protein staining was positive while other hormone tests were negative in human non-functioning pituitary adenomas derived cell-F.Conclusions There is a chromosomal aberration,which forms a chimera in in-vitro cultured non-functioning pituitary adenomas cells.The aberrant chromosome is associated with the proliferation and secretion of pituitary adenoma.