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  • بررسی تاثیر خاصیت ضدسرطانی عصاره گیاه کاکوتی (Ziziphora) بر میزان بیان ژن (CLDN1) Claudin-1 در رده سلول های سرطانی معده (AGS)

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رقم المقالة : JAPAD-2308-1238 زيارة : 152 الصفحة: 29 - 43

نوع المخطوط: ابحاث

بررسی تاثیر خاصیت ضدسرطانی عصاره گیاه کاکوتی (Ziziphora) بر میزان بیان ژن (CLDN1) Claudin-1 در رده سلول های سرطانی معده (AGS)

الموضوعات : مجله پلاسما و نشانگرهای زیستی

زهرا دیلمی خیابانی 1 (گروه زیست شناسی ، دانشکده علوم پایه و فنی و مهندسی، دانشگاه ازاد اسلامی، واحد زنجان، زنجان، ایران)
سارا نیری 2 (گروه زیست شناسی- دانشگده علوم پایه، دانشگاه آزاد اسلامی زنجان، واحد زنجان، زنجان ایران)

تاريخ الإرسال : 25 السبت , محرم, 1445 تاريخ التأكيد : 20 الخميس , ربيع الأول, 1445 تاريخ الإصدار : 08 الإثنين , ربيع الثاني, 1445

الکلمات المفتاحية: 1 claudin-, کاکوتی, سرطان معده, سلول های AGS,

ملخص المقالة :

زمینه وهدف: کلودین ها) (claudin پروتئین های ساختاری و عملکردی اتصالات محکم در سلول های اپیتلیال هستند. الگوهای بیان تغییر یافته از اعضای مختلف claudin در انواع بیماری ها، به ویژه در سرطان ها، به اثبات رسیده است. گزارش شده است که در سرطان معده claudin-1، 22 برابر افزایش بیان را نشان داده است. هدف از این مطالعه بررسی میزان بیان ژن claudin-1 در سلول‌های آدنوکارسینومای معده پس از تیمار سلول‌ها با عصاره گیاهی کاکوتی می باشد. مزیت‌ این عصاره‌ داشتن خاصیت ضد سرطانی و عدم وجود عوارض جانبی می‌باشدمواد و روش ها: سلول‌ی آدنوکارسینومای معده (AGS) در پلیت های 12 چاهکی با 10% FBS و در دمای 37 درجه سلسیوس و انکوباتور 5% CO2 و رطوبت 85% کشت داده شدند. سلول‌ها با غلظت های 800 , 1200 , 2000 μg/ml از عصاره آبی کاکوتی به مدت 48و 72 ساعت تیمار شده سپس استخراج RNA، سنتز cDNA با استفاده از کیت انجام گردید. در نهایت بررسی میزان بیان ژن claudin-1 با پرایمرهای اختصاصی و Real time PCR انجام شده و از ژن GAPDH به عنوان ژن کنترل داخلی استفاده شد. آنالیز داده‌ها با CTΔΔ-2 محاسبه گردید و معنی دار بودن نتایج با T-test بررسی شد.نتایج: آنالیز نتایج نشان داد که در تیمار 48 ساعت سلول‌های AGS با عصاره کاکوتی در غلظت μg/ml 800 و 1200 به ترتیب 7 برابر و 3/2 برابر و در تیمار 72 ساعت، در غلظت μg/ml 800 و 1200 به ترتیب کاهش 5.8 برابر و 10 برابر کاهش در بیان ژن claudin-1 مشاهده گردید و هیچ تغییر معنی داری در غلظت 2000μg/ml در هر دو تیمار مشاهده نشد.نتیجه گیری: با توجه به نتایج حاصل، تاثیر تیمار 72ساعت در سلول های AGS با غلظت‌ μg/ml 1200 عصاره کاکوتی در کاهش بیان ژن 1- claudinبه عنوان ژن درگیر در سرطان‌زایی، بیشتر می‌باشد.در این مطالعه، ژن claudin-1 به‌عنوان یکی از عوامل در بدخیمی تومور با تاثیر عصاره کاکوتی به طور قابل توجهی کاهش بیان را نشان داده است.

المصادر:


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    14.
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    15.
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    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.
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    22.
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    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.
_||_

  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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