JAK抑制剂AG490诱导乳腺癌细胞凋亡及对Survivin表达的影响

Effects of a JAK Inhibitor,AG490,on Proliferation and Survivin Expression of Breast Cancer Cell Line MDA-MB-231

背景与目的:已有研究证实Survivin在多种乳腺癌中可呈高表达且与肿瘤的恶性程度和患者预后密切相关,但是Survivin在乳腺癌细胞中高表达的相关机制尚有待深入了解。本实验拟研究JAK-STAT信号通路抑制剂AG490在人乳腺癌细胞MDA-MB-231中对Survivin表达以及对肿瘤细胞增殖、凋亡的影响。方法:以MDA-MB-231细胞为研究对象,应用AG490后,用Westernblot检测细胞中P-STAT3、Survivin的蛋白表达变化、RT-PCR检测SurvivinmRNA的变化;用MTT法检测细胞增殖活性的改变;用流式细胞术检测细胞凋亡的情况。结果:AG490使MDA-MB-231细胞中P-STAT3和Survivin的蛋白表达减少,同时细胞的增殖活性降低,凋亡增多。P-STAT3/β-actin用药前为0.74,AG49040μmol/L作用24h后为0.21;Survivin/β-actin用药前为1.09,40μmol/LAG490作用24h后为0.28。40μmol/LAG490作用24h后细胞出现“亚G1”峰,凋亡率为5.62%;AG49040μmol/L,作用48h后细胞凋亡率为28.81%。结论:AG490可诱导MDA-MB-231细胞凋亡、减少Survivin的表达。

BACKGROUND ＆ OBJECTIVE. Many researches have proven that Survivin is expressed abundantly and is involved in tumorigenesis and prognosis of breast cancers. However, the signal transduction pathway of Survivin in breast cancer cells is still not clear. This study was to investigate the effects of AG490, a JAK enzyme inhibitor, on the expression of Survivin and on the proliferation and apoptosis of human breast cancer cell line MDA- MB-231. METHODS： MDA-MB-231 cells were treated with AG490. The protein levels of P-STAT3 and Survivin were detected by Western blot; the mRNA level of survivin was measured by reverse transcription-polymerase chain reaction （RT-PCR）. Cell proliferation was detected by MTT;cell apoptosis was detected by flow cytometry （FCM）. RESULTS： AG490 reduced the expression of P-STAT3 and Survivin, inhibited cell proliferation and induced apoptosis in MDA-MB-231 cells. When treated with 40 umol/L AG490 for 24 h, the protein level of P-STAT3 was decreased from 0.74 to 0.21, and that of Survivin was decreased from 1.09 to 0.28. The ＂sub-G1＂ peak appeared and the apoptosis rate was 5.62% at 24 h, and further reached 28.81% at 48 h. CONCLUSION： AG490 could induce apoptosis and reduce the expression of Survivin in breast cancer MDA-MB-231 cells.