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Apoptosis inhibitor 5 gene silenced by miRNA-224 in paclitaxel inducing HepG2 cells apoptosis
SHAN Chang-min* SUN Ye-ying LI Juan WANG Dong YAO Qing-shou TAN Xiu-hua LIU Xiang-yong
Acta Anatomica Sinica ›› 2015, Vol. 46 ›› Issue (4) : 521-527.
Apoptosis inhibitor 5 gene silenced by miRNA-224 in paclitaxel inducing HepG2 cells apoptosis
Objective To explore the gene silencing of microRNAs in paclitaxel inducing HepG2 cells apoptosis.Methods After treated with different concentrations of paclitaxel, apoptosis of human hepotoma cell line HepG2 was detected by flow cytometry, expressions of miR-224 and apoptosis inhibitors 5(api-5) mRNA were determined by Real-time PCR, the expression of API-5 protein was detected by immunohistochemical two steps method and Western blotting method. Results In paclitaxel inducing HepG2 cells apoptosis, the expressions of miR-224 in HepG2 treated with 0.5 mg/L, 1mg/L paclitaxel for 24hours decreased slightly, while those of the others increased obviously by analysis of 2-ΔΔCT method. The expressions of api-5 mRNA in HepG2 treated with 0.5 mg/L, 1mg/L paclitaxel for 24hours decreased slightly, while those of the others were up-regulated except that those of treatment with 0.5mg/L paclitaxel and the control was similar. By the treatment of Image Pro Plus v 5.1,it was found that the levels of API-5 protein were down-regulated with treatment of paclitaxel through the analysis of t-test（P<0.05）. Conclusion Increased miR-224 may affect the expression of its target gene api-5. Through the combination with api-5 mRNA 3’UTR,the completion of translation is prevented, which may impact on liver cancer cell apoptosis pathway.
Paclitaxel / Human hepotoma cell line HepG2 / MicroRNA-224 / Apoptosis inhibitor 5 / Real-time PCR / Western blotting
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