Chinese medicine Icraiin reverses expression of hepatocarcinoma cell line HepG2.2.15 and induces differentiation

Qian Wang, Ling Zhang, Hai-Ting Mao, Hong-Tao Gu, Pei-E Wen, Cui-Ling Li, Institute of Basic Medicine, Shan-dong Academy of Medical Sciences, Key Medical Library for Tumor Immunology & Chinese Medicine Immunology of Shandong Province, Key Library for Modern Medicine and Technology of Shandong Province, Jinan 250062, Shandong Province, ChinaShang-Jun Yang, Institute of Materia Medica, Shandong Academy of Medical Sciences, Jinan 250062, Shandong Province, ChinaWu-Qing Xia, Shandong Qianfoshan Hospital, Jinan 250014, Shandong Province, China  

AIM: To investigate the mechanism of Icraiin (ICA) induction of tumor cell differentiation and apoptosis in HepG2.2.15 cells.METHODS: MTT assay was used to explore the effects of ICA on the proliferation of HepG2.2.15 cells. Reverse transcriptase-polymerase chain reaction was used to examine mRNA levels of P27 and FLIP genes. Flow cytometry was used to detect apoptosis of HepG2.2.15 cells treated by Annexin V-FITC/PI. ABN ProSpec System nephelometer was used to examine alpha fetal protein (AFP), and the Elecsys System was used for immunoassay detection of transferrin (Tf).RESULTS: Treatment with ICA reduced the rate of proliferation in the HepG2.2.15 cell line. The inhibition of cell proliferation was time dependent (P0.05). The distribution ratios of ICA-treated HepG2.2.15 cells were significantly different compared with those of the controls. The number of G0/G1-phase cells increased (56.26 ± 1.56% vs 49.68 ± 1.34%, P 0.01), and Sphase cells decreased (19.95 ± 1.24% vs 28.02 ± 1.03%, P 0.01). ICA increased the relative expression level of P27 mRNA from 0.27 to 0.78, and reduced that of FLIP mRNA from 0.90 to 0.54. ICA also decreased AFP expression level from 285 ± 58 to 156 ± 46 mg/L, while it increased that of Tf from 67.1 ± 24 to 152.1 ± 26 mg/L (P0.05).CONCLUSION: ICA significantly inhibited HepG2.2.15 cell proliferation by affecting the cell cycle distribution ratios, and by reducing the number of S-phase cells and increasing G0/G1-phase cells. ICA could enhance the expression levels of P27 mRNA and Tf, while simultane-ously inhibiting the tumor marker AFP and inducing HepG2.2.15 cell differentiation.

【Key Words】： Icraiin HepG2.2.15 P27 Alpha fetal protein Transferrin Fas-associated death domainlike interleukin-1 beta-converting enzyme inhibitory protein
【Fund】： 国家自然科学基金项目资助 No.30540041
【CateGory Index】： R735.7

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