Lab Anim Res.  2011 Mar;27(1):19-24.

The Apoptotic Effect of the Hexane Extract of Rheum undulatum L. in Oral Cancer Cells through the Down-regulation of Specificity Protein 1 and Survivin

Affiliations
  • 1Department of Oral Pathology, School of Dentistry and Institute of Oral Bioscience, Brain Korea 21 Chonbuk National University, Jeonju, Republic of Korea. efiwdsc@chonbuk.ac.kr
  • 2Department of Preventive Dentistry, School of Dentistry and Institute of Oral Bioscience, Brain Korea 21, Chonbuk National University, Jeonju, Republic of Korea.

Abstract

The hexane extract of Rheum undulatum L. (HERL) has been shown to have anti-cancer activity in several cancers in vivo and in vitro. However, the anti-cancer activity of HERL and its molecular mechanism in human oral cancer cells has not been explored. Thus, the aim of this study was to elucidate the growth-inhibitory and apoptosis-inducing effects of HERL in HN22 and SCC15 oral cancer cell lines. This study shows that HERL inhibits oral cancer growth, decreases cell viability, and causes apoptotic cell death in HN22 and SCC15 cells, as characterized by morphological changes, nuclear condensation and fragmentation, the cleavage of PARP and the accumulation of cells in the sub-G1 phase. The treatment of oral cancer cells with HERL also resulted in decreased expression of specificity protein (Sp1) and its downstream protein, survivin. Therefore, our results suggest that the regulation of Sp1 and survivin plays a critical role in HERL-induced apoptosis in human oral cancer cells.

Keyword

Hexane extract of Rheum undulatum L. (HERL); apoptosis; specificity protein 1; survivin; oral cancer

MeSH Terms

Apoptosis
Cell Death
Cell Line
Cell Survival
Down-Regulation
Humans
Mouth Neoplasms
Rheum
Sensitivity and Specificity*

Figure

  • Figure 1 Morphological changes and the viability of HN22 and SCC15 cells after treatment with hexane extract of Rheum undulatum L. (HERL). A, Photomicrographs of HN22 and SCC15 cells treated with 20, 40 and 60 µg/mL of HERL for 48 h. B, Cell viability was determined using MTS assay. The graph was representative of three independent experiments and bar is mean±SD. *P<0.05 compared to control group.

  • Figure 2 The effect of hexane extract of Rheum undulatum L. (HERL) on cell accumulation in sub-G1 phase. The sub-G1 population was measured by PI staining and flow cytometry analysis. The bar graph represents three independent experiments and mean±SD. *P<0.05 compared to control group.

  • Figure 3 The effect of hexane extract of Rheum undulatum L. (HERL) on the condensation and fragmentation of nucleus in HN22 and SCC15 cells. Fluorescence microscopy images of DAPI-stained HN22 and SCC15 cells showing the appearance of apoptotic morphology.

  • Figure 4 The effect of hexane extract of Rheum undulatum L. (HERL) on the cleavage of poly(ADP-ribose) polymerase (PARP) in HN22 and SCC15 cells. HN22 and SCC15 cells were treated with 20, 40 or 60 µg/mL of HERL for 48 h, and then the protein levels of cleaved PARP were determined by Western blot analysis. Actin was used to normalize the protein loading from each treatment.

  • Figure 5 The effect of hexane extract of Rheum undulatum L. (HERL) on the regulation of Sp1 and survivin protein expression. Sp1 and survivin protein levels were analyzed by Western blot analysis.


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