Effect and mechanism of artemisinin on inhibiting metastasis and invasion of gallbladder carcinoma cells

doi:10.3877/cma.j.issn.2095-3232.2018.05.017

Abstract

Abstract:

Objective

To investigate the effect of artemisinin on the in vitro metastasis and invasion of gallbladder carcinoma cells and its mechanism.

Methods

Gallbladder carcinoma cells GBC-SD was treated with 20 μmol/L artemisinin (artemisinin group) and cells without artemisinin treatment was established as control group. The metastasis ability of gallbladder carcinoma cell was detected using scratch wound assay. The variation of cell invasion was evaluated using Transwell assay. The change of cell morphology was observed using immunofluorescent staining. The mRNA and protein level changes of E-cadherin, Slug, Vimentin, MMP-2 and MMP-9 were detected by qRT-PCR and Western blot. The expression levels between two groups were compared by t test.

Results

At 24 h, the scratch width was (393±23) μm in artemisinin group, significantly wider than (210±15) μm in control group (t=13.415, P<0.05). In artemisinin group, the quantity of transmembrane GBC-SD cells was 93±8, significantly less than 202±14 in control group (t=-17.037, P<0.05). Artemisinin promoted the epithelial-mesenchymal transition of cell morphology. In artemisinin group, the expression levels of Slug, Vimentin, MMP-2 and MMP-9 mRNA of GBC-SD cells were 0.89±0.03, 1.35±0.10, 3.30±0.13 and 1.15±0.08, significantly lower than 2.08±0.12, 3.22±0.15, 4.72±0.19 and 1.95±0.15 in control group (t=-19.812, -20.775, -12.259, -9.438; P<0.05). In artemisinin group, the expression of E-cadherin mRNA was 2.06±0.08, significantly higher than 1.03±0.06 in control group (t=20.956; P<0.05). Western blot showed that artemisinin could down-regulate the expression levels of Slug, Vimentin, MMP-2 and MMP-9 proteins, whereas up-regulate the expression of E-cadherin protein.

Conclusions

Artemisinin can inhibit the metastasis and invasion of gallbladder carcinoma cells. It is probably related with suppressing the expression of Slug gene, reversing the epithelial-mesenchymal transition, thereby down-regulating the expression level of MMP.

Key words: Gallbladder neoplasms, Artemisinins, Cell migration assays, Neoplasm invasiveness

Yiyu Qin, Longyang Jin, Anxing Ge, Zhenyu Hei. Effect and mechanism of artemisinin on inhibiting metastasis and invasion of gallbladder carcinoma cells[J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2018, 07(05): 420-425.

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References 22

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引物名称	正向	反向
E-cadherin	3'-acgatgatgtgaacacc-5'	3'-ttgctgttgtgcttaacccc-5'
Slug	3'-cctggttgcttcaaggacac-5'	3'-tggagcagaggttgttagca-5'
Vimentin	3'-tgcaggctcagattcaggaa-5'	3'-ctccggtactcagtggactc-5'
MMP-2	3'-aatcccaccaaccctcagag-5'	3'-gtgccctcttgagacagtct-5'
MMP-9	3'-gagttcccggagtgagttga-5'	3'-aaaggtgagaagagagggcc-5'

引物名称	正向	反向
E-cadherin	3'-acgatgatgtgaacacc-5'	3'-ttgctgttgtgcttaacccc-5'
Slug	3'-cctggttgcttcaaggacac-5'	3'-tggagcagaggttgttagca-5'
Vimentin	3'-tgcaggctcagattcaggaa-5'	3'-ctccggtactcagtggactc-5'
MMP-2	3'-aatcccaccaaccctcagag-5'	3'-gtgccctcttgagacagtct-5'
MMP-9	3'-gagttcccggagtgagttga-5'	3'-aaaggtgagaagagagggcc-5'

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