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A combination of paclitaxel and siRNA-mediated silencing of Stathmin inhibits growth and promotes apoptosis of nasopharyngeal carcinoma cells

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Abstract

Background

Stathmin, a microtubule associated protein (MAP), is an important molecular target for cancer therapy. Paclitaxel is one of the primary antitumor drugs targeting microtubules (MTs). We hypothesized that decreasing the expression level of Stathmin might improve the effectiveness of paclitaxel in the treatment of nasopharyngeal carcinoma (NPC).

Methods

NPC cell lines, CNE1-LMP1 and HNE2, and a CNE1-LMP1 tumor xenograft mouse model were used to test both in vitro and in vivo our siRNA-based Stathmin silencing strategy. The effects of Stathmin silencing on cell proliferation, apoptosis, and viability were investigated using MTT, AO/EB staining, TUNEL, caspase protein detection, and FCM assays. Cell migration and invasion were assayed using a Transwell assay. The combined effects of Stathmin silencing and paclitaxel were investigated using MTT, FCM, Western blot and indirect immunofluorescence assays. The effect of paclitaxel on Stathmin expression in NPC cells and, in addition, A375, MGC and HeLa cells was determined by RT-PCR and Western blotting.

Results

We found that siRNA-mediated silencing of Stathmin suppresses proliferation, induces apoptosis through the mitochondrial pathway, and causes G2/M-phase cell cycle arrest in the NPC cell lines CNE1-LMP1 and HNE2. Also, the migration and invasion of the respective NPC cells were found to be inhibited. In addition, we show that a combination of Stathmin silencing and paclitaxel is more effective than either agent alone in inhibiting proliferation and inducing apoptosis, cell cycle arrest, and MT polymerization. Furthermore, we found that Stathmin expression in the tumor cells is down-regulated by paclitaxel treatment.

Conclusion

siRNA-mediated silencing of Stathmin suppresses the proliferation, invasion and metastasis, and induces the apoptosis of NPC cells. Paclitaxel reduces the expression of Stathmin, and combining Stathmin silencing with paclitaxel treatment enhances MT polymerization. This combined strategy may provide a new approach for clinical NPC treatment.

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Acknowledgements

This work was supported by the National Basic Research Program of China (973 Program) (2009CB521801 and 2011CB504305).

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Authors and Affiliations

  1. Cancer Research Institute, Xiangya School of Medicine, Central South University, Changsha, 410078, Hunan, People’s Republic of China

    Yong Wu, Min Tang, Yuan Wu, Xinxian Weng, Lifang Yang, Wen Xu, Wie Yi, Jinghe Gao & Ya Cao

  2. Department of Medicine Clinical Laboratory, The Third Xiangya Hospital of Central South University, Changsha, 410013, Hunan, People’s Republic of China

    Yong Wu, Yuan Wu & Wen Xu

  3. Carcinogenesis and Invasion Key Laboratory of Education Ministry of China, Central South University, Changsha, 410078, Hunan, People’s Republic of China

    Yong Wu, Min Tang, Yuan Wu, Xinxian Weng, Lifang Yang, Wen Xu, Wie Yi, Jinghe Gao & Ya Cao

  4. Molecular Image Research Center, Central South University, Changsha, 410078, Hunan, People’s Republic of China

    Yong Wu, Min Tang, Yuan Wu, Xinxian Weng, Lifang Yang, Wen Xu, Wie Yi, Jinghe Gao & Ya Cao

  5. The Hormel Institute, University of Minnesota, Austin, MN, 55912, USA

    Ann M. Bode & Zigang Dong

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  1. Yong Wu
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  2. Min Tang
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Correspondence to Ya Cao.

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Fig. S1

Stathmin expression is silenced by si-stathmin in CNE1-LMP1 cells. CNE1-LMP1 cells were transfected with si-vector, si-Mock (as controls) or si-stathmin. si-Stathmin suppresses the expression of stathmin at the mRNA (a) and protein (b) levels in CNE1-LMP1 cells. The accompanying histogram (right) shows the quantitative luminosity values of each lane individually. Total RNA was extracted 24 h after transfection and was then amplified by RT-PCR. Expression of stathmin mRNA was detected with β-actin as an internal control. Meanwhile, total proteins were extracted 24 h after transfection and then examined by Western blot and α-tublin was used as a loading control. The data are shown as the mean ± S.D. of at least three independent experiments performed in duplication. Asterisks (*) indicate a significant decrease in expression of stathmin induced by si-stathmin. (DOC 571 kb)

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Wu, Y., Tang, M., Wu, Y. et al. A combination of paclitaxel and siRNA-mediated silencing of Stathmin inhibits growth and promotes apoptosis of nasopharyngeal carcinoma cells. Cell Oncol. 37, 53–67 (2014). https://doi.org/10.1007/s13402-013-0163-3

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