表皮生长因子和牛垂体提取物对无血清培养的人垂体腺瘤细胞的作用

Effect of EGF and BPE on the human pituitary adenoma cells cultured in serum-free medium

目的 观察表皮生长因子 (EGF)和牛垂体提取物(BPE)在人垂体腺瘤细胞培养中的作用 .方法 将不同浓度的EGF和BPE作用于体外培养的人垂体腺瘤细胞 ,用MTT及3 H TdR掺入法测定细胞增殖及DNA代谢的变化情况 ,并用TUNEL和台盼蓝染色观察细胞凋亡和死亡情况 .结果 不同浓度的EGF和不同浓度的BPE对体外无血清培养的人垂体腺瘤细胞均表现出明显的刺激作用 ,并呈一定的剂量效应关系 .同时 ,EGF和BPE均可降低体外无血清培养的人垂体腺瘤细胞的死亡及凋亡数量 .以含 10 0mL·L-1血清的DMEM培养液组的A490nm 值和cpm值为 10 0 % ,无血清的DMEM组的A490nm值和cpm值为 (8.6± 5 .2 ) %和 (9.4±5 .4 ) % ;含 2 0mg·L-1EGF和 1mg·L-1BPE的无血清DMEM培养液组的A490nm 值和cpm值为 (98.7± 5 .3) %和(10 2 .4± 5 .5 ) % ,而凋亡指数 (AI)仅为 3.5± 0 .9,提示EGF和BPE之间具有协同作用 .结论 EGF和BPE具有明显改善人垂体腺瘤细胞体外无血清培养条件的作用 。

AIM To observe the effects of epidermal growth factor (EGF) and bovine pituitary extract (BPE) on human pituitary adenoma cells cultured in serum free medium. METHODS After EGF or/and BPE with different concentrations were added to DMEM, MTT method and 3H TdR incorporation were used to examine the changes of proliferation and DNA synthesis of human pituitary tumors cells, and TUNEL and staining with trypan blue were employed to determine the apoptosis or death of human pituitary tumors cells. RESULTS Results from MTT colorimetric determination and 3H TdR incorporation demonstrated that both EGF and BPE of different concentrations had a significant dose dependent stimulatory effect on human pituitary tumors cells cultured with serum free medium in vitro . And results from TUNEL and trypan staining showed that both EGF and BPE of different concentrations decreased the number of apoptosis and death of human pituitary tumors cells cultured with serum free medium in vitro . The values of A 490 nm and cpm of human pituitary tumors cells cultured with 100 mL·L -1 serum medium in vitro were 100% and those of A 490 nm and cpm with serum free medium were (8.6±5.2)% and(9.4±5.4)%. After treatment with EGF (20 mg·L -1 ) and BPE (1 mg·L -1 ) together, the change rate of A 490 nm and cpm was (98.7±5.3)% and (102.4±5.5)% respectively, but that of AI was only 3.5±0.9, indicating that there was a synergistic action between EGF and BPE. CONCLUSION GF and BPE have an important role in human pituitary tumors cells cultured in serum free medium in vitro . EGF and BPE can promote the proliferation of human pituitary tumors cells cultured in vitro .