雷帕霉素对人肺癌细胞株增殖抑制的研究

目的：探讨雷帕霉素对人肺癌细胞株的生长影响及磷酸化核糖体蛋白S6激酶（P-S6K1）的表达在雷帕霉素抑制肺癌细胞增长中的意义。方法：用不同浓度（0、0.1、1.0、10.0、100.0 nmol/L）的雷帕霉素处理人肺癌细胞株,药物敏感试验判断雷帕霉素的敏感株和不敏感株,MTT法检测其对人肺癌细胞增殖的影响。Western blotting观察人肺癌细胞哺乳动物雷帕霉素靶蛋白（mTOR）信号通路上下游蛋白分子及其磷酸化蛋白的表达,比较P-S6K1在用药前后的表达。结果：4个浓度（0.1、1.0、10.0、100.0 nmol/L）的雷帕霉素持续作用72 h,除H1299细胞株外,对HLAMP、A549和PAa细胞株的抑制率相应增加（P〈0.05）,100.0 nmol/L时的抑制率达到最大（P〈0.05）,A值的变化与抑制率一致。位于mTOR上下游的AKT、S6K1、4E-BP1、eIF4E蛋白在所有细胞株中均有表达,P-S6K1在敏感株中呈高表达,在不敏感株中未检测到。在HLAMP和A549中P-S6K1用药后6、12、18和24 h较用药前有所降低。结论：雷帕霉素能抑制人肺癌细胞生长,P-S6K1在一定程度上可代表肺癌细胞株对雷帕霉素的敏感性。

Rapamycin Inhibits Proliferation of Human Lung Cancer Cell Lines

Objective： To investigate the growth inhibition of human lung cancer cell lines in response to rapamycin and the significance of P-S6K1 in its action . Methods： The human lung cancer cell lines were treated with different concentrations （0, 0.1, 1.0, 10.0, 100.0 nmol/L） of Rapamycin. Proliferation of the lung cancer cell lines in response to rapamycin was assessed by MTT assay. Lung cancer cell lines were divided into Rapamycin sensitive and non-sensitive cell lines. The expression and phosphorylation state of multiple components of the mTOR signaling pathway in human lung cancer cells were tested by Western blotting analysis. Expression of P-S6K1 after and before rapamycin was studied. Results： With the exception of H1299, HLAMP, A549 and PAa 3 human lung cancer cell lines were inhibited by varied dose of Rapamycin （0.1, 1.0, 10.0, 100.0 nmol/L） for 72 hours. HLAMP, A549 and PAa were sensitive to Rapamycin （inhibition rate ≥ 30%）, H 1299 was non-sensitive to Rapamycin （inhibition rate〈30%） from 0.1 nmol/L to 100 nmol/L. The change of A was the same as the inhibition rate. roTOR upstream and downstream proteins, including AKT, $6K1,4E-BP1 and eIF4E, were expressed in all cell lines. P-S6K1 was intensely expressed in sensitive cell lines but was not detectable in non-sensitive cell lines. Expression of P-S6K1 in HLAMP and A549 was lowered after 6, 12, 18 and 24 hours from rapamycin use. Conclusion： Rapamycin can inhibit human lung cancer cell growth and the expression of P-S6K1 in human lung cancer cell lines may reflect sensitivity to Rapamycin.