Anticancer effects and underlying mechanism of Colchicine on human gastric cancer cell lines in vitro and in vivo

Tao Zhang, Wei Chen, Xumian Jiang, Lei Liu, Kai Wei, Hansong Du, Hui Wang, Juan Li, Tao Zhang, Wei Chen, Xumian Jiang, Lei Liu, Kai Wei, Hansong Du, Hui Wang, Juan Li

Abstract

The present study investigated the effects of Colchicine on gastric carcinoma (GC) cells and explored its possible mechanisms underlying such effects. The results of MTT and colony formation assays showed that Colchicine (2, 5, and 10 ng/ml) markedly inhibited the proliferation of AGS and NCI-N87 cells in a dose-dependent manner. It also led to a reduction in cell migration in both GC cells as determined by Transwell migration assay. Mover, data form Hoechst 33342 staining and flow cytometry assay indicated that Colchicine (2, 5, and 10 ng/ml) promoted the apoptosis of NCI-N87 cells. In addition, the release of cytochrome c, the activation of bax, and the inhibition of bcl-2 were observed in NCI-N87 cells treated with Colchicine. Furthermore, the in vivo experiment further confirmed that Colchicine administration remarkably suppressed the tumor growth in nude mice via induction of apoptosis at 0.05 and 0.1 mg/kg. In addition, no visible toxicity was observed in liver and renal tissue of mice. This finding suggests that Colchicine-induced apoptosis is associated with caspase-3-mediated mitochondrial apoptotic pathways.

Keywords: Colchicine; apoptosis; caspase-3; gastric cancer; mitochondrial apoptotic pathways.

Conflict of interest statement

The authors declare that there are no competing interests associated with the manuscript.

© 2019 The Author(s).

Figures

Figure 1. The structure of Colchicine
Figure 1. The structure of Colchicine
Figure 2. The effect of Colchicine on…
Figure 2. The effect of Colchicine on cell growth and migration
(A) Effect of Colchicine on the cell viability of human GC AGS and NCI-N87 cells at 48 h. (B) Effect of Colchicine on colony formation of human GC AGS and NCI-N87 cells on day 12. (C) Effect of Colchicine on the migration of human GC AGS and NCI-N87 cells at 48 h. (D) Quantitative analysis of the percentage of migrated cells after treatment with Colchicine at 48 h. Data are means ± S.D. (n=5). **P<0.01, ***P<0.001, compared with control of AGS, ##P<0.01, ###P<0.001, compared with control of NCI-N87.
Figure 3. The effect of Colchicine on…
Figure 3. The effect of Colchicine on cell apoptosis
(A) Morphological analysis of the nuclei of NCI-N87 cells stained by Hoechst 33342 after various concentrations of Colchicine treatment for 48 h (200×). (B) Quantitative analysis of the percentage of apoptotic NCI-N87 cells stained by Hoechst 33342 after various concentrations of Colchicine treatment for 48 h. (C) Flow cytometry analysis of NCI-N87 cells after various concentrations of Colchicine treatment for 48 h. (D) The percentage of apoptotic cells in each group after various concentrations of Colchicine treatment for 48 h. Data are means ± S.D. (n=5). **P<0.01, ***P<0.001, compared with untreated control.
Figure 4. The effect of Colchicine on…
Figure 4. The effect of Colchicine on apoptotic protein expression
(A) The protein expression of bax, bcl-2, cleaved-caspase-3, and cytochrome c in NCI-N87 cells after various concentrations of Colchicine treatment for 48 h. (B) The protein expression of PI3K, p-PI3K, Akt, p-Akt, mTOR, and p-mTOR in NCI-N87 cells after various concentrations of Colchicine treatment for 48 h. Data are means ± S.D. (n=5). *P<0.05, **P<0.01, ***P<0.001, compared with untreated control.
Figure 5. The effect of Colchicine on…
Figure 5. The effect of Colchicine on tumor growth and apoptosis in vivo
(A) The antitumor effects of Colchicine on TV of xenograft model of NCI-N87 cells. (B) The antitumor effects of Colchicine on tumor weight of xenograft model of NCI-N87 cells. (C) Apoptotic cells were detected in xenograft tumor tissue using the TUNEL assay (200×). (D) Quantitated results of the TUNEL assay. Data are means ± S.D. (n=10). *P<0.05, **P<0.01, compared with untreated control.

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