两种酪氨酸激酶抑制剂对结肠癌细胞增殖的影响

The tffects of two tyrosine kinase inhibitors on the proliferation of colon cancer cell

目的:研究两种酪氨酸激酶抑制剂对体外培养的HT - 2 9结肠癌细胞生长的影响及可能机制。方法:用MTT比色法观察不同浓度的Genistein及TyrphostinAG14 78对体外培养的结肠癌细胞生长的影响;免疫细胞化学法检测两种酪氨酸激酶抑制剂(Tyrosinekinaseinhibitors,TKIs)对肿瘤细胞PCNA表达的影响;Westernblot技术分析Genistein及TyrphostinAG14 78对结肠癌细胞ERK - 1及pERK表达的影响。结果:不同浓度的Genistein及TyrphostinAG14 78对HT - 2 9结肠癌细胞的增殖均具有抑制作用,并具有良好的量-效关系。免疫细胞化学检测表明,Genistein及TyrphostinAG14 78明显下调HT- 2 9细胞PCNA表达。蛋白质印迹检测发现,两种酪氨酸激酶抑制剂不抑制ERK表达,但能够抑制ERK磷酸化。结论:酪氨酸激酶抑制剂Genistein及TyrphostinAG14 78能有效抑制HT - 2 9结肠癌细胞生长。

Objective:To investigate the effects of Genistein and Tyrphostin AG1478 on the proliferation of colon cancer cell HT-29 and its possible mechanism.Methods:The effect of two tyrosine kinase inhibitors (TKIs) on HT-29 cell lines was studied by using MTT assay.Immunocytochemical staining was performed with PCNA polyclonal antibody in HT-29 cells.Western blot assay was used to detect the expression of ERK-1 and pERK.Results:Both Genistein and Tyrphostin AG1478 significantly inhibit the growth of HT-29 cells and showed concentration dependence.Two tyrosine kinase inhibitors suppressed PCNA expression.Western blot revealed that Genistein and Tyrphostin not inhibit ERK-1 expression but inhibit phosphorylation of ERK.Conclusion:Tyrosine kinase inhibitors Genistein and Tyrphostin AG1478 can inhibit the growth of HT-29 colon cancer cells,the anti-proliferation mechanism may be related to down-regulate the expression of pERK.