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Manuscript ID : JAPAD-2308-1238 Visit : 128 Page: 29 - 43

Article Type: Original Research

study the effect of Ziziphora Extract on Claudin1 Gene Expression in Cancerous Cell line AGS.

Subject Areas :

Zahra  Deilami Khiabani 1 (Department of Biology, Faculty of Basic Sciences, Zanjan Branch, Islamic Azad University, Zanjan, Iran.)
Sara  Nayyeri 2 (Biology department, Basic Science,Islamic Azad University, Zanjan branch, Zanjan, Iran)

Received: 2023-08-12 Accepted : 2023-10-05 Published : 2023-10-23

Keywords: Ziziphora, Claudin-1, AGS cells, gastric cancer,

Abstract :

Background & Aims: Claudins are the main structural and functional proteins of tight junctions in epithelial cells. The altered expression patterns of differ­ent claudin members have been demonstrated in a variety of diseases, particularly in cancers.  It has been reported that claudin 1 is overexpressed 22-fold in gastric adenocarcinoma.  Considering the least side effect of herbal extracts, in this study we have evaluated  claudin-1 gene expression in AGS cells treated with ziziphora extract.Materials and Methods: The AGS cells were incubated 37°C containing 5% CO2  with 85% humidity DMEM with  10% FBS .  The cells were treated with concentrations of 800, 1200, 2000 μg /ml of ziziphora for 48 and 72 hours. Extraction of RNA, synthesis of cDNA has been done using kit. The study of claudin-1 gene expression was performed by Real time PCR and also GAPDH gene was used as the internal control. Data analysis was performed with 2 - ΔΔCT and statistically analyzed by T-Test  .Results: The results of Real time PCR data have shown in 48 hours treatment, reduction of 7 and 2.3 fold with concentrations of 800 and 1200 μg /ml of ziziphora  respectively  and  in  72 hours treatment, reduction of 5.8 and 10 fold have seen in concentrations of 800 and 1200μg /ml of ziziphora, respectively and there was no significant  change in concentration of 2000μg / ml in both treatments.Conclusion: Ziziphora extract in 1200 μg /ml concentration, reduced the expression of claudin-1 remarkably. Claudin-1 may become therapeutic targets for cancer treatment and ziziphora  as a herbal extract decreased the expression rate of claudin1significantly, in lower concentrations treatment.

References:


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  24. Miwa N, Furuse M, Tsukita S, Niikawa N, Nakamura Y and Furukawa Y. Involvement of claudin-1 in the beta-catenin/ Tcf signaling pathway and its frequent upregulation in human colorectal cancers. Oncol Res. 2001; 12(11-12):469- 476.
    25.
  25.  Jie Huang, et al, Claudin-1 enhances tumor proliferation and metastasis by regulating cell anoikis in gastric cancer. 2014
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  26. Eftang LL, Esbensen Y, Tannaes TM, Blom GP, Bukholm IR and Bukholm G. Up-regulation of CLDN1 in gastric cancer is correlated with reduced survival. BMC Cancer. 2013; 13:586.
    27.
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    28.
  28. Resnick MB, Gavilanez M, Newton E, Konkin T, Bhattacharya B, Britt DE, Sabo E and Moss SF. Claudin expression in gastric adenocarcinomas: a tissue microarray study with prognostic correlation. Hum Pathol. 2005; 36(8):886-892.
    29.
  29. Resnick MB, Gavilanez M, Newton E, et al. Claudin expression in gastric adenocarcinomas: a tissue microarray study with prognostic correlation. Hum Pathol. 2005;36(8):886–892.
    30.
  30. Gyõrffy H, Holczbauer A, Nagy P, et al. Claudin expression in Barrett’s esophagus and adenocarcinoma. Virchows Arch. 2005;447(6): 961–968.
    31.
  31. Hough CD, Sherman-Baust CA, Pizer ES, et al. Large-scale serial analysis of gene expression reveals genes differentially expressed in ovarian cancer. Cancer Res. 2000;60(22):6281–6287.
    32.
  32. Kwon MJ, Kim SH, Jeong HM, et al. Claudin-4 overexpression is associated with epigenetic derepression in gastric carcinoma. Lab Invest. 2011;91(11):1652–1667.
    33.
  33. Hoevel T, Macek R, Swisshelm K, Kubbies M. Reexpression of the TJ protein CLDN1 induces apoptosis in breast tumor spheroids. Int J Cancer. 2004;108(3):374–383.
    34.
  34. Hossein Zadegan, Hassan, Ezzatpour, Abdullahpour, Fawad, Motamedi, Rashidipour. Investigating the cytotoxicity effects of olive and green tea extracts on breast cancer cell line. Journal of Ardabil University of Medical Sciences. 2010 Dec 10;10(4):287-94
    35.
  35. Lars L Eftang, Ying Esbensen , Tone M Tannæs , Gustav P Blom , Ida RK Bukholm and Geir Bukholm : Up-regulation of CLDN1 in gastric cancer is correlated with reduced survival. BMC Cancer 2013, 13:586
    36.
  36. Jung H, Jun KH, Jung JH, Chin HM, Park WB: The expression of claudin-1, claudin-2, claudin-3, and claudin-4 in gastric cancer tissue. J Surg Res 2011, 167:e185–e191.
    37.

 

  1. Zavala-Zendejas VE, Torres-Martinez AC, Salas-Morales B, Fortoul TI, Montano LF, Rendon-Huerta EP: Claudin-6, 7, or 9 overexpression in the human gastric adenocarcinoma cell line AGS increases its invasiveness, migration, and proliferation rate. Cancer Invest 2011, 29:1–11.
    2.
_||_

  1. Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D. Global cancer statistics. CA Cancer J Clin 2011; 61 (2): 69- 90.
    2.
  2. Lichtenstein P, Holm NV, Verkasalo PK, Iliadou A, Kaprio J, Koskenvuo M, et al. Environmental and heritable factors in the causation of cancer--analyses of cohorts of twins from Sweden, Denmark, and Finland. N Engl J Med 2000; 343(2): 78-85
    3.
  3. Gupta GP and Massague´ J. Cancer metastasis: building a framework. Cell 2006;127(4):679-9.
    4.
  4. Valastyan S, Weinberg RA. Tumor metastasis: molecular insights and evolving paradigms. Cell 2011;147(2):275-92.
    5.
  5. Noori-Daloii MR. Medical Molecular Genetics in The Third Millennium. Tehran: Samer publishing; 2009, 2012. [Persian]
    6.
  6. Noori-Daloii MR. Emery’s Elements of Medical genetics. 6th ed. Tehran: Jame-e-Negar and Salemi Publishing; 2012. [Persian]
    7.
  7. Rendon-Huerta E,Teresa F,Teresa GM, Xochitl GS,Georgina AF,et al. (2010) Distribution and expression pattern of claudins 6, 7, and 9 in diffuse- and intestinal-type gastric adenocarcinomas. J Gastrointest Cancer 41: 52-59.
    8.
  8. Jung H, Jun KH, Jung JH, Chin HM, Park WB (2010) The Expression of Claudin-1, Claudin-2, Claudin-3, and Claudin-4 in Gastric Cancer Tissue. J Surg Res.
    9.
  9. Arivusudar Marimuthu and et al. Gene Expression Profiling of Gastric Cancer. J Proteomics Bioinform 2011, 4:4
    10.
  10. Schneeberger EE and Lynch RD. The tight junction: a multifunctional complex. Am J Physiol Cell Physiol. 2004; 286(6):C1213-1228.
    11.
  11. Findley MK and Koval M. Regulation and roles for claudin-family tight junction proteins. IUBMB Life. 2009; 61(4):431-437.
    12.
  12. Lal-Nag M, Morin PJ (2009) The claudins. Genome Biol 10: 235.
    13.
  13. Wu YL, Zhang S, Wang GR and Chen YP. Expression transformation of claudin-1 in the process of gastric adenocarcinoma invasion. World J Gastroenterol. 2008; 14(31):4943-4948.
    14.
  14. Jung H, Jun KH, Jung JH, Chin HM and Park WB. The expression of claudin-1, claudin-2, claudin-3, and claudin-4 in gastric cancer tissue. J Surg Res. 2011; 167(2):e185-191.
    15.
  15. Ichiyasu H, McCormack JM, McCarthy KM, Dombkowski D, Preffer FI and Schneeberger EE. Matrix metalloproteinase-9-deficient dendritic cells have impaired migration through tracheal epithelial tight junctions. American journal of respiratory cell and molecular biology. 2004; 30(6):761-770.
    16.
  16. Yoon CH, Kim MJ, Park MJ, Park IC, Hwang SG, An S, Choi YH, Yoon G and Lee SJ. Claudin-1 acts through c-Abl-protein kinase Cdelta (PKCdelta) signaling and has a causal role in the acquisition of invasive capacity in human liver cells. J Biol Chem. 2010; 285(1):226-233.
    17.
  17.  Leotlela PD, Wade MS, Duray PH, Rhode MJ, Brown HF, Rosenthal DT, Dissanayake SK, Earley R, Indig FE, Nickoloff BJ, Taub DD, Kallioniemi OP, Meltzer P, Morin PJ and Weeraratna AT. Claudin-1 overexpression in melanoma is regulated by PKC and contributes to melanoma cell motility. Oncogene. 2007; 26(26):3846- 3856.
    18.
  18. Frisch SM and Francis H. Disruption of epithelial cellmatrix interactions induces apoptosis. J Cell Biol. 1994; 124(4):619-626.
    19.
  19. Frisch SM and Screaton RA. Anoikis mechanisms. Curr Opin Cell Biol. 2001; 13(5):555-562.
    20.
  20. Taddei ML, Giannoni E, Fiaschi T and Chiarugi P. Anoikis: an emerging hallmark in health and diseases. J Pathol. 2012; 226(2):380-393
    21.
  21. Dhawan P, Singh AB, Deane NG, No Y, Shiou SR, Schmidt C, Neff J, Washington MK and Beauchamp RD. Claudin-1 regulates cellular transformation and metastatic behavior in colon cancer. J Clin Invest. 2005; 115(7):1765-1776.
    22.
  22. Singh AB, Sharma A and Dhawan P. Claudin-1 expression confers resistance to anoikis in colon cancer cells in a Srcdependent manner. Carcinogenesis. 2012; 33(12):2538- 2547.
    23.
  23. Huang J, Li J, Qu Y, Zhang J, Zhang L, Chen X, Liu B and Zhu Z. The expression of claudin 1 correlates with betacatenin and is a prognostic factor of poor outcome in gastric cancer. Int J Oncol. 2014; 44(4):1293-1301.
    24.
  24. Miwa N, Furuse M, Tsukita S, Niikawa N, Nakamura Y and Furukawa Y. Involvement of claudin-1 in the beta-catenin/ Tcf signaling pathway and its frequent upregulation in human colorectal cancers. Oncol Res. 2001; 12(11-12):469- 476.
    25.
  25.  Jie Huang, et al, Claudin-1 enhances tumor proliferation and metastasis by regulating cell anoikis in gastric cancer. 2014
    26.
  26. Eftang LL, Esbensen Y, Tannaes TM, Blom GP, Bukholm IR and Bukholm G. Up-regulation of CLDN1 in gastric cancer is correlated with reduced survival. BMC Cancer. 2013; 13:586.
    27.
  27. Soini Y, Tommola S, Helin H and Martikainen P. Claudins 1, 3, 4 and 5 in gastric carcinoma, loss of claudin expression associates with the diffuse subtype. Virchows Arch. 2006; 448(1):52-58
    28.
  28. Resnick MB, Gavilanez M, Newton E, Konkin T, Bhattacharya B, Britt DE, Sabo E and Moss SF. Claudin expression in gastric adenocarcinomas: a tissue microarray study with prognostic correlation. Hum Pathol. 2005; 36(8):886-892.
    29.
  29. Resnick MB, Gavilanez M, Newton E, et al. Claudin expression in gastric adenocarcinomas: a tissue microarray study with prognostic correlation. Hum Pathol. 2005;36(8):886–892.
    30.
  30. Gyõrffy H, Holczbauer A, Nagy P, et al. Claudin expression in Barrett’s esophagus and adenocarcinoma. Virchows Arch. 2005;447(6): 961–968.
    31.
  31. Hough CD, Sherman-Baust CA, Pizer ES, et al. Large-scale serial analysis of gene expression reveals genes differentially expressed in ovarian cancer. Cancer Res. 2000;60(22):6281–6287.
    32.
  32. Kwon MJ, Kim SH, Jeong HM, et al. Claudin-4 overexpression is associated with epigenetic derepression in gastric carcinoma. Lab Invest. 2011;91(11):1652–1667.
    33.
  33. Hoevel T, Macek R, Swisshelm K, Kubbies M. Reexpression of the TJ protein CLDN1 induces apoptosis in breast tumor spheroids. Int J Cancer. 2004;108(3):374–383.
    34.
  34. Hossein Zadegan, Hassan, Ezzatpour, Abdullahpour, Fawad, Motamedi, Rashidipour. Investigating the cytotoxicity effects of olive and green tea extracts on breast cancer cell line. Journal of Ardabil University of Medical Sciences. 2010 Dec 10;10(4):287-94
    35.
  35. Lars L Eftang, Ying Esbensen , Tone M Tannæs , Gustav P Blom , Ida RK Bukholm and Geir Bukholm : Up-regulation of CLDN1 in gastric cancer is correlated with reduced survival. BMC Cancer 2013, 13:586
    36.
  36. Jung H, Jun KH, Jung JH, Chin HM, Park WB: The expression of claudin-1, claudin-2, claudin-3, and claudin-4 in gastric cancer tissue. J Surg Res 2011, 167:e185–e191.
    37.

 

  1. Zavala-Zendejas VE, Torres-Martinez AC, Salas-Morales B, Fortoul TI, Montano LF, Rendon-Huerta EP: Claudin-6, 7, or 9 overexpression in the human gastric adenocarcinoma cell line AGS increases its invasiveness, migration, and proliferation rate. Cancer Invest 2011, 29:1–11.
    2.

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