DOI QR코드

DOI QR Code

Ethanol Extract of Smilax glabra Induces Apoptotic Cell Death in Human YD10B Oral Squamous Cell Carcinoma Cells

  • Young Sun Hwang (Department of Dental Hygiene, College of Health Science, Eulji University)
  • Received : 2023.08.07
  • Accepted : 2023.08.28
  • Published : 2023.09.30

Abstract

Background: Smilax glabra has various pharmacological activities and is widely used as a herbal medicine. Although the incidence of oral cancer is low, the recurrence rate is high, and the 5-year survival rate is poor. It is necessary to search for anticancer drugs that increase the effect of cancer chemotherapy on heterogeneous oral tissues and reduce the side effects on normal cells. This study aimed to investigate the effects and mechanism of ethanol extract of Smilax glabra (EESG) as an anticancer drug for oral cancer. Methods: Smilax glabra root components extracted with 70% ethanol were used to analyze their effects on cancer cells. A 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide assay was performed for cytotoxicity analysis. Flow cytometry was performed to determine the cell cycle phase distribution. To observe apoptotic cells, terminal deoxynucleotidyl transferase dUTP nick end labeling and γH2AX were detected by fluorescence microscope. The protein levels of cleaved PARP and caspase were analyzed using western blotting. The activation of procaspase-3 was confirmed by measuring caspase-3 activity. Results: EESG was no cytotoxic to normal gingival fibroblast but was high in YD10B oral squamous cell carcinoma (OSCC) cells. EESG treatment increased the subdiploid DNA content of YD10B cells by assessing DNA content distribution. Chromatin condensation and DNA strand breaks increased in YD10B cells treated with EESG. EESG-treated YD10B cells had high Annexin V and low propidium iodide levels, confirming that early apoptosis was induced. In addition, increased levels of γH2AX foci, a marker of DNA damage, were observed in the nuclei of EESG-treated YD10B cells. The EESG-treated YD10B cells also exhibited decreased procaspase-3 and procaspase-9 levels, increased PARP cleavage and caspase-3 activity. Conclusion: These results indicate that EESG inhibited cancer cell proliferation by inducing apoptosis in YD10B OSCC cells.

Keywords

Acknowledgement

This research was supported by Eulji University in 2023 and the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2022R1F1A1063204).

References

  1. Li CQ, Lei HM, Hu QY, Li GH, Zhao PJ: Recent advances in the synthetic biology of natural drugs. Front Bioeng Biotechnol 9: 691152, 2021. https://doi.org/10.3389/fbioe.2021.691152 
  2. Kwak SW: Research trends in the development of new drugs and medicines based on natural products. Biological Research Information Center. Retrieved May 1, 2023, from https://www.khidi.or.kr/board/view?pageNum=1&rowCnt=10&no1=&linkId=48775229&menuId=MENU01845&maxIndex=00487752299998&minIndex=00487356519998&schType=0&schText=&schStartDate=&schEndDate=&boardStyle=&categoryId=&continent=&country= (2019, June 11). 
  3. Wu H, Wang Y, Zhang B, et al.: Smilax glabra Roxb.: a review of its traditional usages, phytochemical constituents, pharmacological properties, and clinical applications. Drug Des Devel Ther 16: 3621-3643, 2022. https://doi.org/10.2147/DDDT.S374439 
  4. Chen L, Lan Z, Zhou Y, et al.: Astilbin attenuates hyperuricemia and ameliorates nephropathy in fructose-induced hyperuricemic rats. Planta Med 77: 1769-1773, 2011. https://doi.org/10.1055/s-0030-1271135 
  5. Chen L, Yin H, Lan Z, et al.: Anti-hyperuricemic and nephroprotective effects of Smilax china L. J Ethnopharmacol 135: 399-405, 2011. https://doi.org/10.1016/j.jep.2011.03.033 
  6. Wu LS, Wang XJ, Wang H, Yang HW, Jia AQ, Ding Q: Cytotoxic polyphenols against breast tumor cell in Smilax china L. J Ethnopharmacol 130: 460-464, 2010. https://doi.org/10.1016/j.jep.2010.05.032 
  7. Xu W, Liu J, Li C, Wu HZ, Liu YW: Kaempferol-7-O-beta-D-glucoside (KG) isolated from Smilax china L. rhizome induces G2/M phase arrest and apoptosis on HeLa cells in a p53-independent manner. Cancer Lett 264: 229-240, 2008. https://doi.org/10.1016/j.canlet.2008.01.044 
  8. Li YL, Gan GP, Zhang HZ, et al.: A flavonoid glycoside isolated from Smilax china L. rhizome in vitro anticancer effects on human cancer cell lines. J Ethnopharmacol 113: 115-124, 2007. https://doi.org/10.1016/j.jep.2007.05.016 
  9. Hua S, Zhang Y, Liu J, et al.: Ethnomedicine, phytochemistry and pharmacology of Smilax glabra: an important traditional Chinese medicine. Am J Chin Med 46: 261-297, 2018. https://doi.org/10.1142/S0192415X18500143 
  10. Ban JY, Cho SO, Koh SB, Song KS, Bae K, Seong YH: Protection of amyloid beta protein (25-35)-induced neurotoxicity by methanol extract of Smilacis chinae rhizome in cultured rat cortical neurons. J Ethnopharmacol 106: 230-237, 2006. https://doi.org/10.1016/j.jep.2005.12.034 
  11. Guo Y, Mao W, Jin L, et al.: Flavonoid group of Smilax glabra Roxb. regulates the anti-tumor immune response through the STAT3/HIF-1 signaling pathway. Front Pharmacol 13: 918975, 2022. https://doi.org/10.3389/fphar.2022.918975 
  12. Kim MY, Kim CS, Lee SH, Kim JW, Jang HJ: A clinicostatistical analysis of oral cancer patients for recent 8 years. J Korean Assoc Oral Maxillofac Surg 33: 660-668, 2007. 
  13. Han JG, Kim SI, Park B, Jang JY, Shin YS, Kim CH: A study on the clinicopathological characteristics associated with cervical lymph mode metastasis and extra-nodal extension in patients with oral cancer. Korean J Head Neck Oncol 37: 33-41, 2021. https://doi.org/10.21593/kjhno/2021.37.2.33 
  14. Davidson M, Chau I, Cunningham D, et al.: Impact of tumour histological subtype on chemotherapy outcome in advanced oesophageal cancer. World J Gastrointest Oncol 9: 333-340, 2017. https://doi.org/10.4251/wjgo.v9.i8.333 
  15. Chalertpet K, Sangkheereeput T, Somjit P, Bankeeree W, Yanatatsaneejit P: Effect of Smilax spp. and Phellinus linteus combination on cytotoxicity and cell proliferation of breast cancer cells. BMC Complement Med Ther 23: 177, 2023. https://doi.org/10.1186/s12906-023-04003-x 
  16. She T, Qu L, Wang L, et al.: Sarsaparilla (Smilax Glabra Rhizome) extract inhibits cancer cell growth by S phase arrest, apoptosis, and autophagy via redox-dependent ERK1/2 pathway. Cancer Prev Res (Phila) 8: 464-474, 2015. https://doi.org/10.1158/1940-6207.CAPR-14-0372 
  17. Mohan C, Long K, Mutneja M, Ma J: Detection of end-stage apoptosis by ApopTag TUNEL technique. Methods Mol Biol 1219: 43-56, 2015. https://doi.org/10.1007/978-1-4939-1661-0_5 
  18. Podhorecka M, Skladanowski A, Bozko P: H2AX phosphorylation: its role in DNA damage response and cancer therapy. J Nucleic Acids 2010: 920161, 2010. https://doi.org/10.4061/2010/920161 
  19. Gregory CD, Devitt A: The macrophage and the apoptotic cell: an innate immune interaction viewed simplistically? Immunology 113: 1-14, 2004. https://doi.org/10.1111/j.1365-2567.2004.01959.x 
  20. McIlwain DR, Berger T, Mak TW: Caspase functions in cell death and disease. Cold Spring Harb Perspect Biol 5: a008656, 2013. https://doi.org/10.1101/cshperspect.a008656 Erratum in: Cold Spring Harb Perspect Biol 7: a026716, 2015. https://doi.org/10.1101/cshperspect.a026716 
  21. Chaitanya GV, Steven AJ, Babu PP: PARP-1 cleavage fragments: signatures of cell-death proteases in neurodegeneration. Cell Commun Signal 8: 31, 2010. https://doi.org/10.1186/1478-811X-8-31 
  22. Gao Y, Su Y, Qu L, et al.: Mitochondrial apoptosis contributes to the anti-cancer effect of Smilax glabra Roxb. Toxicol Lett 207: 112-120, 2011. https://doi.org/10.1016/j.toxlet.2011.08.024 
  23. Ooi LS, Wong EY, Chiu LC, Sun SS, Ooi VE: Antiviral and anti-proliferative glycoproteins from the rhizome of Smilax glabra Roxb (Liliaceae). Am J Chin Med 36: 185-195, 2008. https://doi.org/10.1142/S0192415X08005692