Contradictory Effects of 6-Shogaol on the Human Cervical Cancer Cell Line HeLa Through Network Analysis

Document Type : Original paper


1 Cell Therapy and Regenerative Medicine Research Center, Endocrinology and Metabolism Molecular-Cellular Sciences Institute, Tehran University of Medical Sciences, Tehran, Iran.

2 Proteomics Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

3 Surgery Department, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran.

4 Traditional Medicine and Materia Medica Research Center and Department of Traditional Medicine, School of Traditional Medicine Shahid Beheshti University of Medical Sciences, Tehran, Iran.

5 Critical Care Quality Improvement Research Center, Faculty of Paramedical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

6 Laser Application in Medical Sciences Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.


Background and objectives: To identify new targets for cancer clinical management, protein-protein interaction (PPI) network analysis of proteome data could accelerate this approach. For this aim, proteins with differential expressions in 6-shogaol exposure from proteomics study underwent protein-protein interaction (PPI) network analysis. Methods: Cytoscape version 3.8.2 and its plug-ins including NetworkAnalyzer and ClueGO2.5.8+CluePedia1.5.8 were applied for the construction and the corresponding analysis of the network. Results: A number of six differentially expressed proteins (DEPs) were identified as hub-bottlenecks of the PPI network. The critical proteins GAPDH, ENO1, HSP90AB1, ACTG1, RPSA, and CALR were determined as central elements of the analyzed network and their related biological processes were identified as “protein folding chaperones” and “glucose catabolic process”. Conclusion: Potential candidates may be predicted as both anticancer agents and promoters of side effects of 6-shogaol in cancer treatment; however, complementary studies are required to provide validation and deeper understating of its molecular behavior in this regard.  


Main Subjects

  • Luo H, Vong CT, Chen H, Gao Y, Lyu P, Qiu L, Zhao M, Liu Q, Cheng Z, Zou J, Yao P, Gao C, Wei J, Ung COL, Wang S, Zhong Z, Wang Naturally occurring anti-cancer compounds: shining from Chinese herbal medicine. Chin Med. 2019; 14(1): 1–58.
  • Safarzadeh E, Shotorbani SS, Baradaran B. Herbal medicine as inducers of apoptosis in cancer treatment. Adv Pharm Bull. 2014; 4(S1): 421–427.
  • Anh NH, Kim SJ, Long NP, Min JE, Yoon YC, Lee EG, Kim M, Kim TJ, Yang YY, Son EY, Yoon SJ, Diem NC, Kim HM, Kwon Ginger on human health: a comprehensive systematic review of 109 randomized controlled trials. Nutrients. 2020; 12(1): 1–28.
  • Singletary K. Ginger: an overview of health benefits. Nutr Today. 2010; 45(4): 171–183.
  • Ma RH, Ni ZJ, Zhu YY, Thakur K, Zhang F, Zhang YY, Hu F, Zhang JG, Wei A recent update on the multifaceted health benefits associated with ginger and its bioactive components. Food Funct. 2021; 12(2): 519–542.
  • Lei D, Hong T, Li L, Chen L, Luo X, Wu Q, Liu Isobaric tags for relative and absolute quantitation‑based proteomics analysis of the effect of ginger oil on bisphenol A‑induced breast cancer cell proliferation. Oncol Lett. 2021; 21(2): 1–13.
  • Jeena K, Liju VB, Kuttan R. Antitumor and cytotoxic activity of ginger essential oil (Zingiber officinale Roscoe). Int J Pharm Pharm Sci. 2015; 7(8): 341–344.
  • Prasad S, Tyagi AK. Ginger and its constituents: role in prevention and treatment of gastrointestinal cancer. Gastroenterol Res Pract. 2015; Article ID 142979.
  • Liao DW, Cheng C, Liu JP, Zhao LY, Huang DC, Chen GT. Characterization and antitumor activities of polysaccharides obtained from ginger (Zingiber officinale) by different extraction methods. Int J Biol Macromol. 2020; 152: 894–903.
  • Sarip MSM, Morad NA, Ali NAM, Yusof YAM, Yunus MAC. The kinetics of extraction of the medicinal ginger bioactive compounds using hot compressed water. Sep Purif Technol. 2014; 124: 141–147.
  • Ling H, Yang H, Tan SH, Chui WK, Chew EH. 6‐Shogaol, an active constituent of ginger, inhibits breast cancer cell invasion by reducing matrix metalloproteinase‐9 expression via blockade of nuclear factor‐κB activation. Br J Pharmacol. 2010; 161(8): 1763–1777.
  • Ha SK, Moon E, Ju MS, Kim DH, Ryu JH, Oh MS, Kim 6-Shogaol, a ginger product, modulates neuroinflammation: a new approach to neuroprotection. Neuropharmacology. 2012; 63(2): 211–223.
  • Pan MH, Hsieh MC, Kuo JM, Lai CS, Wu H, Sang S, Ho 6‐Shogaol induces apoptosis in human colorectal carcinoma cells via ROS production, caspase activation, and GADD 153 expression. Mol Nutr Food Res. 2008; 52(5): 527–537.
  • Tan BS, Kang O, Mai CW, Tiong KH, Khoo ASB, Pichika MR, Bradshaw TD, Leong 6-Shogaol inhibits breast and colon cancer cell proliferation through activation of peroxisomal proliferator activated receptor γ (PPARγ). Cancer Lett. 2013; 336(1): 127–139.
  • Saha A, Blando J, Silver E, Beltran L, Sessler J, Di Giovanni J. 6-Shogaol from dried ginger inhibits growth of prostate cancer cells both in vitro and in vivo through inhibition of STAT3 and NF-κB signaling. Cancer Prev Res. 2014; 7(6): 627–638.
  • Kim MO, Lee MH, Oi N, Kim SH, Bae KB, Huang Z, Kim DJ, Reddy K, Lee SY, Park SJ, Kim JY, Xie H, Kundu JK, Ryoo ZY, Bode AM, Surh YJ, Dong [6]-Shogaol inhibits growth and induces apoptosis of non-small cell lung cancer cells by directly regulating Akt1/2. Carcinogenesis. 2014; 35(3): 683–691.
  • Mukkavilli R, Yang C, Tanwar RS, Saxena R, Gundala SR, Zhang Y, Ghareeb A, Floyd SD, Vangala S, Kuo WW, Rida PCJ, Aneja Pharmacokinetic- pharmacodynamic correlations in the development of ginger extract as an anticancer agent. Sci Rep. 2018; 8(1): 1–10.
  • Zhang MM, Wang D, Lu F, Zhao R, Ye X, He L, Ai L, Wu Identification of the active substances and mechanisms of ginger for the treatment of colon cancer based on network pharmacology and molecular docking. Bio Data Min. 2021; 14(1): 1–16.
  • Liu Y, Whelan RJ, Pattnaik BR, Ludwig K, Subudhi E, Rowland H, Claussen N, Zucker N, Uppal S, Kushner DM, Felder M, Patankar MS, Kapur Terpenoids from Zingiber officinale (ginger) induce apoptosis in endometrial cancer cells through the activation of p53. PLoS One. 2012; 7(12): 1–10.
  • Pashaei-Asl R, Pashaei-Asl F, Gharabaghi PM, Khodadadi K, Ebrahimi M, Ebrahimie E, Pashaiasl The inhibitory effect of ginger extract on ovarian cancer cell line; application of systems biology. Adv Pharm Bull. 2017; 7(2): 241–249.
  • Weinreb O, Amit T, Youdim MB. A novel approach of proteomics and transcriptomics to study the mechanism of action of the antioxidant–iron chelator green tea polyphenol (-)-epigallocatechin-3-gallate. Free Radic Biol Med. 2007; 43(4): 546–556.
  • Zamanian Azodi M, Rezaei Tavirani M, Esmaeili S, Arjmand B, Jahani Sherafat S. Evaluation of anticancer effect of ghost pepper: a bioinformatics assessment. Res J Pharmacogn. 2021; 8(3): 77–82.
  • Ji Q, Zhu F, Liu X, Li Q, Su SB. Recent advance in applications of proteomics technologies on traditional Chinese medicine research. Evid Based Complement Alternat Med. 2015; Article ID 983139.
  • Azodi MZ, Arjmand B, Razzaghi M, Tavirani MR, Ahmadzadeh A, Rostaminejad M. Platelet and haemostasis are the main targets in severe cases of COVID-19 infection; a system bioinformatics study. Arch Acad Emerg Med. 2021; 9(1): 1–9.
  • Zamanian Azodi M, Esmaeili S, Rezaei Tavirani M. Bioinformatics identification of green tea anticancer properties: a network-based approach. Res J Pharmacogn. 2021; 8(2): 17–25.
  • Vu M, Yu J, Awolude OA, Chuang L. Cervical cancer worldwide. Curr Probl Cancer. 2018; 42(5): 457–465.
  • Small Jr W, Bacon MA, Bajaj A, Chuang LT, Fisher BJ, Harkenrider MM, Jhingran A, Kitchener HC, Mileshkin LR, Viswanathan AN, Gaffney Cervical cancer: a global health crisis. Cancer. 2017; 123(13): 2404–2412.
  • Cohen PA, Jhingran A, Oaknin A, Denny L. Cervical cancer. 2019; 393(10167): 169–182.
  • Akinyemiju TF. Socio-economic and health access determinants of breast and cervical cancer screening in low-income countries: analysis of the world health survey. PLoS One. 2012; 7(11): 1–8.
  • Liu Q, Peng YB, Qi LW, Cheng XL, Xu XJ, Liu LL, Liu EH, Li The cytotoxicity mechanism of 6-shogaol-treated HeLa human cervical cancer cells revealed by label-free shotgun proteomics and bioinformatics analysis. Evid Based Complement Altern Med. 2012; Article ID 278652.
  • Cline MS, Smoot M, Cerami E, Kuchinsky A, Landys N, Workman C, Christmas R, Avila-Campilo I, Creech M, Gross B, Hanspers K, Isserlin R, Kelley R, Killcoyne S, Lotia S, Maere S, Morris J, Ono K, Pavlovic V, Pico AR, Vailaya A, Wang PL, Adler A, Conklin BR, Hood L, Kuiper M, Sander C, Schmulevich I, Schwikowski B, Warner GJ, Ideker T, Bader Integration of biological networks and gene expression data using Cytoscape. Nat Protoc. 2007; 2(10): 2366–2382.
  • Doncheva NT, Morris JH, Gorodkin J, Jensen LJ. Cytoscape StringApp: network analysis and visualization of proteomics data. J Proteome Res. 2018; 18(2): 623–632.
  • Assenov Y, Ramírez F, Schelhorn SE, Lengauer T, Albrecht M. Computing topological parameters of biological networks. 2008; 24(2): 282–284.
  • Esmaeili S, Rostami-Nejad M, Rezaei Tavirani M, Okhovatian F, Zadeh-Esmaeel MM, Razzagh Z, Ahmadzadeh A, Vafaee Evaluating of gene expression alteration after garlic consumption, analyzing through bioinformatics approach. Iran J Pharm Res. 2021; 20(1): 72–81.
  • Bindea G, Galon J, Mlecnik B. CluePedia Cytoscape plugin: pathway insights using integrated experimental and in silico data. Bioinformatics. 2013; 29(5): 661–663.
  • Bindea G, Mlecnik B, Hackl H, Charoentong P, Tosolini M, Kirilovsky A, Fridman WH, Pagès F, Trajanoski Z, Galon ClueGO: a Cytoscape plug-in to decipher functionally grouped gene ontology and pathway annotation networks. Bioinformatics. 2009; 25(8): 1091–1093.
  • Hung JY, Hsu YL, Li CT, Ko YC, Ni WC, Huang MS, Kuo 6-Shogaol, an active constituent of dietary ginger, induces autophagy by inhibiting the AKT/mTOR pathway in human non-small cell lung cancer A549 cells. J Agric Food Chem. 2009; 57(20): 9809–9816.
  • Dugasani S, Pichika MR, Nadarajah VD, Balijepalli MK, Tandra S, Korlakunta JN. Comparative antioxidant and anti-inflammatory effects of [6]-gingerol, [8]-gingerol, [10]-gingerol and [6]-shogaol. J Ethnopharmacol. 2010; 127(2): 515–520.
  • Gholizadeh E, Rezaei Tavirani M, Emadi A, Karbalaei R, Khaleghian A. A new drug discovery approach based on thermal proteome profiling to develop more effective drugs. Middle East J Rehabil Health. 2021; 8(2): 1–9.
  • Zhang JY, Zhang F, Hong CQ, Giuliano AE, Cui XJ, Zhou GJ, Zhang GJ, Cui Critical protein GAPDH and its regulatory mechanisms in cancer cells. Cancer Biol Med. 2015; 12(1): 10–22.
  • Colell A, Green D, Ricci J. Novel roles for GAPDH in cell death and carcinogenesis. Cell Death Differ. 2009; 16(12): 1573–1581.
  • Guo C, Liu S, Sun MZ. Novel insight into the role of GAPDH playing in tumor. Clin Transl Oncol. 2013; 15(3): 167–172.
  • Kim JW, Kim SJ, Han SM, Paik SY, Hur SY, Kim YW, Lee JM, Namkoong Increased glyceraldehyde-3-phosphate dehydrogenase gene expression in human cervical cancers. Gynecol Oncol. 1998; 71(2): 266–269.
  • Li HJ, Ke FY, Lin CC, Lu MY, Kuo YH, Wang YP, Liang KH, Lin SC, Chang YH, Chen HY, Yang PC, Wu HC. ENO1 promotes lung cancer metastasis via HGFR and WNT signaling-driven epithelial-mesenchymal transition. Cancer Res. 2021; 81(15): 4094–4109.
  • Gou Y, Li F, Huo X, Hao C, Yang X, Pei Y, Li N, Liu H, Zhu ENO1 monoclonal antibody inhibits invasion, proliferation and clone formation of cervical cancer cells. Am J Cancer Res. 2021; 11(5): 1946–1961.
  • Yu L, Shen J, Mannoor K, Guarnera M, Jiang F. Identification of ENO1 as a potential sputum biomarker for early-stage lung cancer by shotgun proteomics. Clin Lung Cancer. 2014; 15(5): 372–378.
  • Zhou J, Zhang S, Chen Z, He Z, Xu Y, Li Z. CircRNA-ENO1 promoted glycolysis and tumor progression in lung adenocarcinoma through upregulating its host gene ENO1. Cell Death Dis. 2019; 10(12): 1–14.
  • Wang H, Deng G, Ai M, Xu Z, Mou T, Yu J, Liu H, Wang S, Li Hsp90ab1 stabilizes LRP5 to promote epithelial–mesenchymal transition via activating of AKT and Wnt/β-catenin signaling pathways in gastric cancer progression. Oncogene. 2019; 38(9): 1489–1507.
  • Kontostathi G, Zoidakis J, Makridakis M, Lygirou V, Mermelekas G, Papadopoulos T, Vougas K, Vlamis-Gardikas A, Drakakis P, Loutradis D, Vlahou A, Anagnou N, Pappa Cervical cancer cell line secretome highlights the roles of transforming growth factor-beta-induced protein ig-h3, peroxiredoxin-2, and NRF2 on cervical carcinogenesis. Biomed Res Int. 2017; Article ID 4180703.
  • Dong X, Han Y, Sun Z, Xu J. Actin gamma 1, a new skin cancer pathogenic gene, identified by the biological feature‐based classification. J Cell Biochem. 2018; 119(2): 1406–1419.
  • Yan Y, Xu H, Zhang L, Zhou X, Qian X, Zhou J, Huang Y, Ge W, Wang RRAD suppresses the warburg effect by downregulating ACTG1 in hepatocellular carcinoma. Onco Targets Ther. 2019; 12: 1691–1703.
  • Richter C, Mayhew D, Rennhack JP, So J, Stover EH, Hwang JH, Szczesna-Cordary Genomic amplification and functional dependency of the gamma actin gene actg1 in uterine cancer. Int J Mol Sci. 2020; 21(22): 1–15.
  • Chen S, Wang X, Yuan J, Deng C, Xie X, Kang J. Reduced levels of actin gamma 1 predict poor prognosis in ovarian cancer patients. J Obstet Gynaecol Res. 2020; 46(9): 1827–1834.
  • Song DG, Kim YS, Jung BC, Rhee KJ, Pan CH. Parkin induces upregulation of 40S ribosomal protein SA and posttranslational modification of cytokeratins 8 and 18 in human cervical cancer cells. Appl Biochem Biotechnol. 2013; 171(7): 1630–1638.
  • Wu Y, Tan X, Liu P, Yang Y, Huang Y, Liu X, Meng X, Yu B, Wu M, Jin ITGA6 and RPSA synergistically promote pancreatic cancer invasion and metastasis via PI3K and MAPK signaling pathways. Exp Cell Res. 2019; 379(1): 30–47.
  • Fucikova J, Spisek R, Kroemer G, Galluzzi L. Calreticulin and cancer. Cell Res. 2021; 31(1): 5–16.
  • Wang K, Li H, Chen R, Zhang Y, Sun XX, Huang W, Bian H, Chen Combination of CALR and PDIA3 is a potential prognostic biomarker for non-small cell lung cancer. Oncotarget. 2017; 8(57): 96945–96957.
  • Sun L, Liu M, Sun GC, Yang X, Qian Q, Feng S, Mackey LV, Coy Notch signaling activation in cervical cancer cells induces cell growth arrest with the involvement of the nuclear receptor NR4A2. J Cancer. 2016; 7(11): 1388–1395.
  • Zhu H, Zhu H, Tian M, Wang D, He J, Xu T. DNA methylation and hydroxymethylation in cervical cancer: diagnosis, prognosis and treatment. Front Genet. 2020; 11(347): 1–12.