| OBJECTIVE: To investigate the antitumor mechanism of triptolide based on hn RNPA2 / B1-PI3K-Akt signaling pathway.Method:To detect the effect of triptolide on the proliferation of lung cancer cell line A549 by CCK-8 assay.Cell scratch assay was used to detect the inhibitory effect of triptolide on A549 cell proliferation.The effect of triptolide on the cell cycle and apoptosis of lung cancer A549 were detected by flow cytometry.Western blot was used to detect the effect of triptolide on the expression of proteins including Hn RNPA2/B1,PI3 K,AKT which belong to the HNRNP A2/ B1-PI3K-AKT signaling pathway.The effect of triptolide on the expression of PI3 K,AKT and related genes Hn RNPA2/B1,PI3 K,AKT was detected by real-time quantitative PCR.RESULTS: After treated with triptolide(10,20,40,50,100 nm)for 24 h,the IC50 of triptolide on A549 cells was calculated to be 50 nm by CCK-8 toxicity test.The healing area of the blank group was significantly lower than 100 nm triptolide group after 0,24 and 48 h cell scratch test.However,after treated with 50 nm and 100 nm triptolide for 24 h,the apoptosis rates of A549 cells were 8.2% and 22.8%respectively by flow cytometry,and the expression of hn RNPA2 / B1,PI3 K and Akt decreased gradually in A549 cells which detected by q PCR.Meanwhile,the western blotting showed that the expression of hn RNPA2 / B1,PI3 K and Akt were decreasing with the increase of triptolide dose.In the tumor model experiment which A549 lung cancer cell line was subcutaneously injected into nude mice,the results of q PCR and Western blotting showed that the expression of hn RNPA2 / B1,PI3 K,Akt and other genes were inhibited and the expression of hn RNPA2 / B1,PI3 K,Akt and other proteins were down regulated after triptolide treatment.CONCLUSIONS: Triptolide can inhibit the proliferation of non-small cell lung cancer cells,the mechanism of which may be through inhibiting the expression of hn RNPA2/B1 in tumor cells,and then inhibiting the expression of PI3 K,Akt and related proteins in PI3K/ Akt signaling pathway,so as to induce the apoptosis of tumor cells and play an against NSCLC... |
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