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. 2015 Jun 11:15:59.
doi: 10.1186/s12935-015-0210-4. eCollection 2015.

Effect of Matrine on HPAC cell migration by down-regulating the expression of MT1-MMP via Wnt signaling

Yongchao Ma #  1   2 , Fazhang Zou #  1   2 , Junping Xiong  1 , Wei Wan  1 , Li Yin  1 , Xianjia Li  1 , Zhanyu Bei  3 , Lei Yuan  1 , Song Meng  1 , Jianguo Wang  1 , Guohua Song  1
Affiliations

Effect of Matrine on HPAC cell migration by down-regulating the expression of MT1-MMP via Wnt signaling

Yongchao Ma et al. Cancer Cell Int. .

Abstract

Aim: This study sought to explore the exact mechanism of Matrine inhibited migration and invasion of human pancreatic cancer cells.

Methods: HPAC or Capan-1 cells were cultured in completed RPMI-1640 medium, contained with 50 μg/ml Matrine or 0.05 μg/ml docetaxel, respectively. Cell viability was evaluated by spectrophotometric analysis using MTT assay. Wound healing assay and transwell approach were used to detect the effects of Matrine on HPAC cell migration and invasion. Western Blot and RT-PCR were performed to detect the expressions of MT1-MMP, Wnt and β-Catenin. CHIP assay was used to detect whether the MT1-MMP transcription activity correlated with Wnt signaling pathway.

Results: MTT results indicated that cell proliferration was inhibited by Matrine at a range of concentrations, especially at high dose. We further found that Matrine treatment significantly induced cell migration and invasion decreased. Interestingly, the expression of MT1-MMP decreased evidently upon Matrine treatment, paralleled with the expressions of Wnt and β-Catenin detected by Western Blot and RT-PCR assay. Further analysis of MT1-MMP transcription activity revealed that Matrine reduced the expression of MT1-MMP mediated by Wnt signaling pathway.

Conclusion: Matrine play a vital role in inhibiting HPAC cellular migration and invasion through down-regulating the expression of MT1-MMP via Wnt signaling pathway.

Keywords: Cell migration; Human pancreatic cancer; MT1-MMP; Matrine; Wnt.

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Figures

Fig. 1
Fig. 1
The migration of HPAC cells was inhibited by Matrine. Log-phase cells were treated with normal complete RPMI-1640 alone or contained with 50 μg/ ml Matrine or 0.05 μg/ ml Docetaxel (a). Data were expressed as mean ± S.E.M from three separated experiments (b). Cell invasion ability was detected by transwell assay (c). Statistical analyses was performed using the t-test. * (P < 0.05) indicates a significant difference compared with the control group
Fig. 2
Fig. 2
Matrine reduced the mRNA expression of MT1-MMP in HPAC cells. The mRNA expression of MT1-MMP in HPAC cell was analyzed by RT-PCR (a). The mRNA of GAPDH was used for internal control, that indicated the equal total mRNA. Data were expressed as mean ± S.E.M from three independent experiments. * (P < 0.05) indicates a significant difference compared with the control group (b)
Fig. 3
Fig. 3
Effects of Matrine on the expressions of MMP2 and MMP9 in HPAC cells. HPAC cells were treated as described previously. The concentrations of MMP2 (a), MMP9 (b) in cell culture supernatant were analysed with ELISA assay. * (p < 0.05) indicates a significant difference compared with the control group
Fig. 4
Fig. 4
Effects of Matrine on the expressions of MT1-MMP, Wnt, β-catenin in HPAC cells. HPAC cells were treated as described previously. The expressions of Wnt, β-catenin and MT1-MMP were detected with western blot. Equal loading proteins were shown with β-actin immunoblot (a). The transcription activity of MT1-MMP in HPAC cells were detected by CHIP assay (b)
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