HER2/neu抗体对SKBR3乳腺癌细胞株酪氨酸磷酸化及细胞增殖的影响

目的研究抗ＨＥＲ２／ｎｅｕ胞外区的特异性抗体对ＳＫＢＲ３乳腺癌细胞株酪氨酸磷酸化及细胞增殖的影响。方法Ｗｅｓｔｅｒｎ印迹法及抗体介导的特异性细胞增殖试验；筛选乳腺癌病人抗ＨＥＲ２／ｎｅｕ抗体阳性血清ＩｇＧ的ＥＬＩＳＡ方法。结果试验发现单抗ｃ－ｎｅｕ－５对ＳＫＢＲ３细胞ｐ１８５酪氨酸磷酸化有明显促进作用，但对细胞增殖的影响不明显；而单抗ｃ－ｎｅｕ－２对ＳＫＢＲ３细胞ｐ１８５酪氨酸磷酸化及细胞增殖均有明显作用。在此基础上，收集乳腺癌病人血清１４份，分离纯化其血清ＩｇＧ，并经ＥＬＩＳＡ方法筛选出５份乳腺癌病人抗ＨＥＲ２／ｎｅｕ抗体阳性血清ＩｇＧ。细胞增殖试验发现该５份标本中有３份对ＳＫＢＲ３细胞增殖有明显抑制作用。选其中１份抑制增殖的阳性血清ＩｇＧ进一步研究其对ＳＫＢＲ３细胞ｐ１８５酪氨酸磷酸化的影响，结果发现该病人血清ＩｇＧ对细胞酪氨酸磷酸化也有抑制作用。结论抗ＨＥＲ２／ｎｅｕ的特异性抗体可能通过抑制细胞的信号传递系统而抑制肿瘤生长。

Effects of specific antibodies against the extracellular domain of the HER2/neu on tyrosine phosphorylation and cell prolifreation of SKBR3 breast cancer cell line

Effects of specific McAbs against the extracellular domain of the HER2/neu on tyrosine phosphorylation and cell proliferation of SKBR3 breast cancer cell line were analyzed with Western blot technique and antibody dependent specific cell proliferation assay .The results showed that McAb c neu 5 enhanced tyrosine phosphorylation of p185 and had no distinct function on SKBR3 cell proliferation,yet McAb c neu 2 inhibited both tyrosine phosphorylation and cell proliferation.Based on these experiments, we further collected 14 sera of breast cancer patients,separated and purified the sera IgG.5 Anti HER2/neu positive sera IgG were further selected from patients with breast cancer with ELISA method. Cell proliferation assay showed that 3 of 5 sera IgG could inhibit cell proliferation of SKBR3. Then we selected 1 positive sera IgG which could inhibit cell proliferation ,to test cell tyrosine phosphorylation of SKBR3. The result showed that this patient sera IgG could also inhibit tyrosine phosphorylation.Therefore, It suggests that the possible mechanism of the inhibition of tumor growth by anti HER2/neu specific antibody probably through interfering with the activation of signal transduction system.