目的分析比较侵袭性和非侵袭性垂体腺瘤原代细胞培养中成纤维细胞的增殖及凋亡的差别,探讨成纤维细胞在垂体腺瘤侵袭性生长中的作用。方法分别对10例侵袭性和10例非侵袭性垂体腺瘤组织进行原代和传代细胞培养,对成纤维细胞进行形态学及细胞增殖速度观察,免疫组织化学染色法检测成纤维细胞Ⅰ型胶原和波形蛋白的表达,比较两组成纤维细胞的DNA合成代谢,流式细胞仪(FCM)检测两组成纤维细胞凋亡。结果侵袭性垂体腺瘤原代细胞培养过程中。第1—2天即可见到成纤维细胞,到第8天左右几乎长满单层,非侵袭性垂体腺瘤原代细胞培养过程中,第10天左右才可见到极少数的成纤维细胞生长,1个月后长满单层,成纤维细胞呈Ⅰ型胶原及波形蛋白阳性表达,侵袭组成纤维细胞的DNA合成量明显高于非侵袭组成纤维细胞(t=28.28,P〈0.05),侵袭组细胞凋亡占细胞总数的(1.4±0.4)%,非侵袭组凋亡细胞占总数的(5.8±1.2)%,两组差别有统计学意义(t=11.00,P〈0.05)。结论侵袭性垂体腺瘤中的成纤维细胞增殖快.凋亡少.垂体腺瘤的侵袭性生长可能与成纤维细胞的增殖有关。
Objective To analyze and compare the difference of growth and apoptosis of the fibroblasts derived from pituitary adenoma with andwithout invasion. To study the role of fibrobalsts in the invasive growth of the pituitary adenoma. Methods Ten invasive pituitary adenoma tissue and ten non-invasive pituitary adenoma tissue were cultured. To observe the changes in cell morphology and cell growth velocity, Immunohistochemical technique was used to detect the expression of collagen Ⅰ and vimentin of the fibroblasts. To compare the difference of DNA constructive metabolism, and to detect the apoptosis of the fibroblasts with flow eytometer, Results In the primary cell culture of the invasive pituitary adenoma, the fibroblasts appeared at 1-2 d, covered the bottom of the culture flask at 8 d. The fibroblasts appeared after 10 day's culture, covered the bottom of the culture flask after 1 month in the primary cell culture of the invasive pituitary adenoma, collagen Ⅰ and vimentin were positive expression in the fibroblasts. The DNA constructive metabolism of the fibroblasts derived from invasive pituitary adenoma was higher than that of the fibroblasts in the pituitary adenoma without invasion ( t = 28.28, P 〈 0.05). The apoptosis rate (1.4 ± 0.4) % of the the fibroblasts derived from invasive pituitary adenoma was lower than that (5.8 ± 1.2) % of the fibroblasts in the pituitary adenoma without invasion ( t = 11.00, P 0.05) .Conclusion The proliferation rate of fibroblasts in invasive pituitary adenoma was higher than that of fibroblasts in non-invasive pituitary adenoma, and the apoptosis rate of fibroblasts in invasive pituitary adenoma was few. The overgrowth of the fibroblasts of the pituitary adenoma may be benifieial to the invasive growth of the pituitary adenoma.