Effects and mechanism of Lanthanum Citrate on the proliferation and apoptosis of hepatocellular carcinoma cell line SMMC-7721.

Abstract

BACKGROUND/AIMS:

To investigate the effect and the possible mechanism of lanthanum citrate on the proliferation and apoptosis of human hepatocellular carcinoma (HCC) cell line SMMC-7721 through the Hedgehog (Hh) signaling pathway.

MATERIALS AND METHODS:

Different concentrations of lanthanum citrate and KAAD-cyclopamine (the Hh signaling pathway representative inhibitor) were used to treat SMMC-7721 cells. Cell proliferation was detected using Methylthiazolyldiphenyl-tetrazolium bromide (MTT) assays. Cell apoptosis was detected using flow cytometry analysis of Annexin V-FITC/ propidium iodide (PI). The protein expressions of regulatory genes, such as cell cycle protein D1 (CyclinD1), cyclin-dependent kinase inhibitor 1 (p21), cysteinyl aspartate specific proteinase 3 (Caspase-3), B-cell lymphoma-2 (Bcl-2), glioma-associated oncogene homolog 1 (Gli1), and sonic hedgehog (Shh) were quantified using Western blot assays. The mRNA expressions of Gli1 and Shh were tested using quantitative real-time polymerase chain reaction (qRT-PCR) assays and the protein expressions of Gli1 and Shh were determined using immunofluorescence assays.

RESULTS:

The Annexin V-FITC and PI double staining results revealed that the 0.1 mM lanthanum citrate group and the 15 µM KAAD-cyclopamine group had both increased the apoptosis rate of SMMC-7721 cells. Both lanthanum citrate and KAAD-cyclopamine downregulated the protein expressions of CyclinD1, Bcl-2, Gli1, and Shh and upregulated the protein expressions of p21 and Caspase-3. Additionally, the immunofluorescence results revealed that the protein expressions of Gli1 and Shh were significantly decreased in both the lanthanum citrate group and the KAAD-cyclopamine group compared to the control group.

CONCLUSION:

Lanthanum citrate inhibits proliferation and promotes apoptosis in HCC SMMC-7721 cells by suppressing the Hh signaling pathway.

• Authors: Bin Chen, Zeming Hu, Biao Chen, Bofei Li
• Journal: The Turkish journal of gastroenterology : the official journal of Turkish Society of Gastroenterology (Turk J Gastroenterol) Vol. 31 Issue 3 Pg. 264-271 (03 2020) ISSN: 2148-5607 [Electronic] Turkey
• PMID: 32343239 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• 3-keto-N-aminoethylaminoethylcaproyldihydrocinnamoyl cyclopamine
• Cinnamates
• Citrates
• Hedgehog Proteins
• Veratrum Alkaloids
• lanthanum citrate

Topics

• Apoptosis (drug effects)
• Carcinoma, Hepatocellular (drug therapy)
• Cell Line, Tumor
• Cell Proliferation (drug effects)
• Cinnamates (pharmacology)
• Citrates (pharmacology)
• Hedgehog Proteins (antagonists & inhibitors, metabolism)
• Humans
• Liver Neoplasms (drug therapy)
• Signal Transduction (drug effects)
• Veratrum Alkaloids (pharmacology)