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Metastasis-induction and apoptosis-protection by TWIST in gastric cancer cells

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Abstract

TWIST, a basic helix-loop-helix transcription factor, has been recently reported to play an important role in tumorigenesis of human cancer through converting the early stage tumors into invasive malignancies. Upregulation of TWIST is often found in cancer patients, especially those with shorter survival period and poor response to chemotherapy. Here we studied the functions of TWIST on regulating migration rate, apoptosis, and gene expression in gastric cancer cells. TWIST expression is elevated in MGC-803 and HGC-27 cells that exhibit high invasive potential; whereas it is reduced in BGC-823 and SGC-7901 cells that possess relatively low invasive content. To evaluate functional consequences of TWIST induction, we examined the effect of TWIST on cell migration and apoptosis. Overexpression of TWIST in BGC-823 cells resulted in increased migration content and decreased sensitivity to the arsenic oxide-induced cell death. Moreover, small interference RNA-mediated TWIST ablation in MGC-803 and HGC-27 cells showed suppressed migration ability, increased induction of apoptosis in response to arsenic oxide, and elevated cell cycle arrest. Furthermore, we found a negative correlation between the TWIST level and p53 level, probably due to transcriptional regulation. Our results have identified TWIST as a critical regulator of gastric cancer cell proliferation and migration, suggesting a potential therapeutic approach to inhibit the growth and metastasis of gastric cancer through inactivation of TWIST.

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Abbreviations

bHLH:

Basic helix-loop-helix

EMT:

Epithelial mesenchymal transition

FCM:

Flow cytometry

MET:

Mesenchymal epithelial transition

PI:

Propidium iodide

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Acknowledgments

We thank Dr. Yunfu Lin (Baylor College of Medicine, USA) for preparation of this manuscript. This research was supported by the fund (LRB04-232) from Postdoctoral Science Foundation of Heilongjiang Province, the fund (LC08C04) of Scientific Research Foundation for Returned Scholars of Heilongjiang Province and the fund (2003AA9CS188-2) from Science and Technology key plan projects of Harbin.

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Authors and Affiliations

  1. The Pathology Department, Affiliated Tumor Hospital of Harbin Medical University, Harbin, China

    Mei-yan Feng, Hong-tao Song, Yu Qin, Qing-tao Shi & Jing-shu Geng

  2. The Abdominal Surgery Department, Affiliated Tumor Hospital of Harbin Medical University, Harbin, China

    Kuan Wang

  3. Department of Histology and Embryology, Harbin Medical University, Harbin, China

    Hong-wei Yu

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  1. Mei-yan Feng
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Correspondence to Jing-shu Geng.

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M.-y. Feng and Kuan Wang were equal contributors to this paper.

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10585_2009_9291_MOESM1_ESM.tif

Supplementary material 1 TWIST knockdown results in decreased invasion and anti-apoptosis ability in HGC-27 cells. All of the experiment procedures were same as Figure 4. A, Wound healing assay showed reduced migrating ability of TWIST depleted cells. B, Representative photograph of transwell assays showed reduced invasion ability brought by TWIST depletion. Invasion assays for untreated cells and for cells treated with ShScramble, ShTWIST1, or ShTWIST2, were carried out in parallel. Invasion ability for untreated cells is not different from cells treated with ShScramble and not shown. C, Decreased apoptosis resistant ability of TWIST depleted cells. D, The relative width of the wounds (%) for ShScramble, ShTWIST1 and ShTWIST2 cells were: 61 ± 8.7, 86 ± 12.5 and 74 ± 2.3 at 12 h after wounding; and 19 ± 8.1, 49 ± 10.2 and 46 ± 4.0 (*p < 0.05, compared with ShScramble) at 24 h after wounding respectively. E, Quantization of the results from triplicate assays. The number of ShScramble, ShTWIST1 and ShTWIST2 cells moved through the membranes was 77.4 ± 16.15, 12.6 ± 3.53 and 21 ± 4.57 respectively. ** indicates p < 0.01 compared with the number for ShScramble cells. F, The proportion of non-viable apoptotic and necrotic cells in total amount of ShScramble, ShTWIST1 and ShTWIST2 cells was 7.05 ± 0.45, 9.55 ± 1.26 and 8.67 ± 0.87 respectively (*p < 0.05 compared with ShScramble). And the ratio of each viable apoptotic cells was 2.62 ± 0.23, 3.51 ± 0.26 and 3.5 ± 0.39 (*p < 0.05 compared with ShScramble). Error bars, ± SD (TIFF 1720 kb)

10585_2009_9291_MOESM2_ESM.tif

Supplementary material 2 TWIST knockdown induces cell cycle arrest. This document is a complement of Figure 5 (TIFF 1226 kb)

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Feng, My., Wang, K., Song, Ht. et al. Metastasis-induction and apoptosis-protection by TWIST in gastric cancer cells. Clin Exp Metastasis 26, 1013–1023 (2009). https://doi.org/10.1007/s10585-009-9291-6

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