Survivin selective inhibitor YM155 induce apoptosis in SK-NEP-1 Wilms tumor cells

BMC Cancer

Jian Pan²  Jian Wang²  Jian Ni⁶  Yan-Hong Li²  Xing Feng²  Na Wang²  Dong Wu²  Li Pang²  Pei-Fang Xiao²  Lan Cao²  Liang Peng⁴  Xuan Zhao³  Li-Chao Sun¹  Xiao-Juan Du⁵  Jun Lu²  Yan-Fang Tao² 

[1]Department of Cell and Molecular Biology, Cancer Institute (Hospital), Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, China

[2]Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou, China

[3]Beijing Insititute for Drug Control, Beijing, China

[4]Institute of Clinical Medical Science, China-Japan Friendship Hospital, Beijing, China

[5]Department of Gastroenterology, the 5th Hospital of Chinese PLA, Yin chuan, Ningxia Province, China

[6]Translational Research Center, Second Hospital, The Second Clinical School, Nanjing Medical University, Nanjing, China

关键词: Real-time PCR array;    Apoptosis;    Survivin;    SK-NEP-1;    YM155;   

Others  :  1127574
DOI  :  10.1186/1471-2407-12-619

 received in 2012-08-29, accepted in 2012-12-21,  发布年份 2012

【 摘 要 】

Background

Survivin, a member of the family of inhibitor of apoptosis proteins, functions as a key regulator of mitosis and programmed cell death. YM155, a novel molecular targeted agent, suppresses survivin, which is overexpressed in many tumor types. The aim of this study was to determine the antitumor activity of YM155 in SK-NEP-1 cells.

Methods

SK-NEP-1 cell growth in vitro and in vivo was assessed by MTT and nude mice experiments. Annexin V/propidium iodide staining followed by flow cytometric analysis was used to detect apoptosis in cell culture. Then gene expression profile of tumor cells treated with YM155 was analyzed with real-time PCR arrays. We then analyzed the expression data with MEV (Multi Experiment View) cluster software. Datasets representing genes with altered expression profile derived from cluster analyses were imported into the Ingenuity Pathway Analysis tool.

Results

YM155 treatment resulted in inhibition of cell proliferation of SK-NEP-1cells in a dose-dependent manner. Annexin V assay, cell cycle, and activation of caspase-3 demonstrates that YM155 induced apoptosis in SK-NEP-1 cells. YM155 significantly inhibited growth of SK-NEP-1 xenografts (YM155 5 mg/kg: 1.45 ± 0.77 cm³; YM155 10 mg/kg: 0.95 ± 0.55 cm³) compared to DMSO group (DMSO: 3.70 ± 2.4 cm³) or PBS group cells (PBS: 3.78 ± 2.20 cm³, ANOVA P < 0.01). YM155 treatment decreased weight of tumors (YM155 5 mg/kg: 1.05 ± 0.24 g; YM155 10 mg/kg: 0.72 ± 0.17 g) compared to DMSO group (DMSO: 2.06 ± 0.38 g) or PBS group cells (PBS: 2.36 ± 0.43 g, ANOVA P < 0.01). Real-time PCR array analysis showed between Test group and control group there are 32 genes significantly up-regulated and 54 genes were significantly down-regulated after YM155 treatment. Ingenuity pathway analysis (IPA) showed cell death was the highest rated network with 65 focus molecules and the significance score of 44. The IPA analysis also groups the differentially expressed genes into biological mechanisms that are related to cell death, cellular function maintenance, cell morphology, carbohydrate metabolism and cellular growth and proliferation. Death receptor signaling (3.87E-19), TNFR1 signaling, induction of apoptosis by HIV1, apoptosis signaling and molecular mechanisms of cancer came out to be the top four most significant pathways. IPA analysis also showed top molecules up-regulated were BBC3, BIRC3, BIRC8, BNIP1, CASP7, CASP9, CD5, CDKN1A, CEBPG and COL4A3, top molecules down-regulated were ZNF443, UTP11L, TP73, TNFSF10, TNFRSF1B, TNFRSF25, TIAF1, STK17A, SST and SPP1, upstream regulator were NR3C1, TP53, dexamethasone , TNF and Akt.

Conclusions

The present study demonstrates that YM155 treatment resulted in apoptosis and inhibition of cell proliferation of SK-NEP-1cells. YM155 had significant role and little side effect in the treatment of SK-NEP-1 xenograft tumors. Real-time PCR array analysis firstly showed expression profile of genes dyes-regulated after YM155 treatment. IPA analysis also represents new molecule mechanism of YM155 treatment, such as NR3C1 and dexamethasone may be new target of YM155. And our results may provide new clues of molecular mechanism of apoptosis induced by YM155.

【 授权许可】

   
2012 Tao et al.; licensee BioMed Central Ltd.

【 图 表 】

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