汉黄芩素调节JAK/STAT信号通路并诱导肺癌A549细胞凋亡及周期阻滞的作用及机制研究

目的研究汉黄芩素对肺癌A549细胞凋亡、周期及对JAK/STAT信号通路的调节作用。方法取对数生长期的肺癌A549细胞,采用低(20μmol/L)、中(40μmol/L)、高(80μmol/L)剂量的汉黄芩素分别干预24 h、48、72 h,MTT法测定不同浓度汉黄芩素对肺癌A549细胞的增殖抑制率,实时定量PCR检测各组肺癌A549细胞中JAK2、STAT3mRNA水平,AnnexinV-FITC/PI双染法检测汉黄芩素对A549细胞凋亡的影响,流式细胞术检测汉黄芩素对A549细胞周期的影响,Western blot检测汉黄芩素对A549细胞JAK2、STAT3水平的影响。结果与对照组相比,汉黄芩素各剂量组肺癌A549细胞24 h、48、72 h增殖抑制率、p-Chk1、P53、Bax水平显著升高(P<0.05),不同浓度的汉黄芩素作用于肺癌A549细胞48 h后,CyclinB1、PCNA、Bcl-2、Survivin、JAK2及STAT3 mRNA水平显著降低(P<0.05),A549细胞凋亡率、G2/M期细胞比例显著升高(P<0.05),Western blot结果表明,汉黄芩素各剂量组A549细胞JAK2及STAT3水平显著降低(P<0.05)。结论汉黄芩素能够抑制肺癌A549细胞的增殖及迁移能力,诱导肺癌A549细胞凋亡及G2/M周期阻滞,其机制可能与调节JAK/STAT信号通路有关。

Effect and mechanism of wogonin on regulating JAK/STAT signal pathway and inducing apoptosis and cell cycle arrest in lung cancer A549 cells

Objective To study the regulatory effect of wogonin on apoptosis,cell cycle and JAK/STAT signal pathway in lung cancer A549 cells.Methods The lung cancer A549 cells in logarithmic growth phase were treated with low(20μmol/L),medium(40μmol/L)and high(80μmol/L)wogonin for 24,48 h and 72 h,respectively.The proliferation inhibition rate of wogonin on lung cancer A549 cells was determined by MTT assay,and the mRNA levels of JAK2 and STAT3 in A549 cells were detected by real-time quantitative PCR.AnnexinV-FITC/PI double staining was used to detect the effect of wogonin on apoptosis of A549 cells.Flow cytometry was used to detect the effect of wogonin on A549 cell cycle,and Western blot was used to detect the effect of wogonin on the levels of JAK2 and STAT3 in A549 cells.Results Compared with the control group,the proliferation inhibition rate of A549 cells treated with wogonin at 24 h,48 h and 72 h,and the level of p-Chk1,P53,Bax was significantly higher(P<0.05).After the A549 cells were treated with wogonin at different concentrations for 48 h,JAK2 and STAT3 mRNA levels of A549 cells were significantly decreased(P<0.05),while the apoptosis rate and G2/M phase ratio of A549 cells were significantly increased(P<0.05).The results of Western blot showed that the levels of CyclinB1,PCNA,Bcl-2,Survivin,JAK2 and STAT3 in A549 cells decreased significantly in each dose group of wogonin(P<0.05).Conclusion Wogonin can inhibit the proliferation and invasion of lung cancer A549 cells,induce apoptosis and G2max M cycle arrest of lung cancer A549 cells,and its mechanism may be related to the regulation of JAK/STAT signal pathway.