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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 the difference of proliferation and apoptosis of fibroblasts between primary and non-invasive pituitary adenomas and to investigate the role of fibroblasts in the invasive growth of pituitary adenomas. Methods Primary and subcultured cells were cultured in 10 cases of invasive and 10 non-invasive pituitary adenomas respectively. Morphology and proliferation rate of fibroblasts were observed. Immunohistochemical staining was used to detect fibroblast type Ⅰ collagen And vimentin expression. The DNA synthesis and metabolism of fibroblasts in two groups were compared. The apoptosis of fibroblasts in both groups was detected by flow cytometry (FCM). Results In primary cultured pituitary adenomas, primary fibroblasts were observed on the first or second day. In the first eight days, almost all primary monolayer and non-invasive primary pituitary adenoma cells were cultured. A few fibroblasts could be seen in about 10 days. After 1 month, the cells were covered with monolayer. The fibroblasts were positive for type I collagen and vimentin. The amount of DNA synthesis of invasive fibroblasts was significantly higher than that of non-invasive ones (1.4 ± 0.4)% of the total cells in the invasive group and (5.8 ± 0.05)% in the non-invasive group (t = 28.28, P <0.05) 1.2%), the difference between the two groups was statistically significant (t = 11.00, P <0.05). Conclusions Fibroblasts in invasive pituitary adenomas proliferate rapidly and have less apoptosis. The invasive growth of pituitary adenomas may be related to the proliferation of fibroblasts.