بررسی اثرات سایتوتوکسیک ناندرولون بر رده سلولی سرطان کولون (HCT) و ارزیابی فعالیت کاسپازهای 3، 8 و 99

فصلنامه زیست شناسی جانوری

دوره 13، شماره 3، خرداد 1400، صفحه 15-25   

نوع مقاله: مقالات پژوهشی

نویسندگان

زهرا اعلایی¹؛ عبدالحسین شیروی^{* 1}؛ رحیم احمدی²؛ ویدا حجتی¹

¹گروه زیست شناسی، واحد دامغان، دانشگاه آزاد اسلامی، دامغان، ایران

²گروه فیزیولوژی، واحد همدان، دانشگاه آزاد اسلامی، همدان، ایران

چکیده

سرطان کولون از مهمترین سرطان­ها در سراسر دنیا محسوب می­شود و سومین سرطان شایع در مردان و دومین سرطان شایع در زنان می­باشد. مطالعات نشان داده­اند که هورمون­های استروئیدی جنسی بر عملکرد دستگاه گوارش و بر تکثیر سلول­های سرطانی در سطح سلولی و مولکولی تاثیرگذارند. این مطالعه به بررسی اثرات ضد تکثیری ناندرولون بر رده سلولی سرطان کلون (HCT) و ارزیابی فعالیت کاسپازهای 3، 8 و 9 می­پردازد. در این مطالعه آزمایشگاهی سلول­های HCT به گروه شاهد (بدون تیمار) و گروه­های تیمار شده با غلظت­های625/0، 25/1، 5/2، 5 و 10 میلی­گرم بر میلی‌لیتر ناندرولون تقسیم­بندی شدند. اثر سیتوتوکسیک هورمون با استفاده از سنجش MTT اندازه­گیری شد. میزان فعالیت کاسپازهای 3، 8 و 9 توسط کیت (Novex) مورد ارزیابی قرار گرفت. زنده­مانی سلول های HCT در گروه دریافت کننده­ی غلظت­های 5/2، 5 و 10 میلی‌گرم بر میلی‌لیتر ناندرولون نسبت به گروه شاهد دچار کاهش معناداری شد (001/0p <) در حالی که فعالیت کاسپاز 3 و 9 (001/0p <) و 8 (01/0 < p) در رده­ سلولی HCT تیمار شده با ناندرولون، افزایش معناداری پیدا کرد. یافته­های این تحقیق نشان دادند که ناندرولون دارای اثر سایتوتوکسیک بر روی سلول­های سرطانی کولون می­باشد و بخشی از این اثر حداقل از طریق افزایش فعالیت کاسپازهای 3 و 8 و 9 حاصل می­شود.

کلیدواژه‌ها

ناندرولون؛ HCT؛ زنده‌مانی؛ کاسپاز 3؛ کاسپاز 8؛ کاسپاز 9

مراجع

1- Acharya P.C., Bansal R. 2013. Synthesis and Antiproliferative Activity of Some Androstene Oximes and Their O‐Alkylated Derivatives. Arch. Pharm, 347: 193–199.

2- Ajduković J.J., Djurendić E.A., Petri E.T., Klisurić O.R, Ćelić A.S., Sakač M.N., Dimitar S.J., Katarina M.P.G. 2013. 17(E)-picolinylidene androstane derivatives as potential inhibitors of prostate cancer cell growth: antiproliferative activity and molecular docking studies. Bioorg Med. Che, 21: 7257-7266.

3- Ajduković J.J., PenovGaši K.M., Jakimov D.S., Klisuric, O.R., Jovanović-Šanta S.S., Sakac M.N., Aleksic L.D., Djurendic E.A. 2015. Synthesis, structural analysis and antitumor acti-vity of novel 17α-picolyl and 17(E)-picolinylidene A-modified androstane derivatives. Bioorg. Med. Chem, 23: 1557-1568.

 4- Alexaki V.I., Charalampopoulos I., Kampa M., Nifli A.P., Hatzogou A., Gravanis A., Castanas E. 2006. Activation of membrane estrogen receptors induce pro-survival kinases, J Steroid Biochem Mol Biol, 98: 97-110.

5- Amani N., Shariati M., Ahmadi R., Khatamsaz S., Mokhtari M. 2019. The Cytotoxic Effects of Testosterone on Gastric Cancer Cell Line (AGS) and Evaluation of Caspase-3, -8, and -9 Activity. Qom Univ Med Sci Journal, 13(6): 9-17. [Persian].

6 - Benten W.P., Lieberherr M., Giese G., Wrehlke C., Stamm O., Sekeris C.E. 1999. Functional testosterone receptors in plasma membranes of T cells. FASEB J, 13:123–133.

7- Campisi J., Yaswen P. 2009. Aging and cancer cell biology, Aging Cell, 8(3) : 221-225.

8- Chen R., Cui J., Wang Q., Li P., Liu X., HU H., Wei W. 2015.  Antiprolifrative effects of anastrozole on MCF-7 human breast cancer cells invitro are significantly enhanced by combind treatment with testosterone undecanoate. Mol Med Rep, 12(1): 769-75.

9- Cohen G.M. 1997. caspase: the executioners of apoptosis, Biochem J, 15; 326(Pt 1): 1–16.

10- Cui J., Liu L., Zhao D., Gan C., Huang X., Xiao Q., Binbin Qi., Lei., Huang Y. 2015. steroids Synthesis, characterization and antitumor activities of some steroidal derivatives with side chain of 17-hydrazone aromatic heterocycle. Steroids, 95: 32.3.

11-  Dai W.S., Kuller L.H., LaPorte R.E., Gutai J.P., Falvo-Gerard L., and Caggiula, A. 1981. The epidemiology of plasma testosterone levels in middle-aged men. Am. J. Epidemiol, 114: 804–816.

12- Degree V.S., Boyce- MYung J. 2003. A Decade of caspase, Oncogene, 22(53): 8543-8567.

13- Fatehm S.H., Amini M., 2008. An epidemiologic study of colorectal cancer in Arak during. Iranian J Surg, 16(2):11-7.

14- Finaly- Schultz J., Sartorius C.A. 2015.  Steroid Hormones, Steroid Receptors, and Breast Cancer Stem Cells. J Mammary Gland Biol Neoplasia, 20: 39-50.

15- Fujita E., Egashira J., Urase K., Kuitda K., Momoi T., 2001. Caspase-9 processing by caspase-3 via a feedback amplification loop in vivo. Cell Death Differ, 8: 335-344.

16- Gatson J.W., Kaur P., Singh M., Gatson JW., Kaur P., Singh M. 2006. Dihydrotestosterone differentially modulates the mitogen-activated protein kinase and the phosphoinositide 3-Kinase/AKt pathways through the unclear and novel membrane androgen receptor in C6 cells. Endocrinology, 147(4): 2028-2034.

17-  Green D.R. 2000.  Apoptotic pathway, paper wrasp stone blynts scissors, Cell, 102: 1-4.

18- Grodstein F., Martinez E., Elizabet A., Platez E.A., Giovannucci E., Cloditz G.A., Kautzky m., Fuchs c., Stampfer M J. 1998. Postmenopausal Hormone Use and Risk for Colorectal Cancer and Adenoma. Ann, Intern, Med, 128: 705-712.

19- Gyovai A., Minorics R.., Kiss A., Mernyák E., Schneider G., Szekeres A.,  Kerekes E., Ocsovszki I., Zupko I. 2018. Antiproliferative Properties of Newly Synthesized 19-Nortestosterone Analogs Without Substantial Androgenic Activity. Front Pharmacol. 9: 825.

20- Hajra K.M., Liu J.R. 2004. Apoptosome dysfunction inhuman cancer. Apoptosis, 9: 691-704.

21 - Hatzogluo A., Kampa M., Kogia C., Charalampopoulos I., Theodoropoulos P.A., Aezinis P., Dambaki C., Papakonstanti E.A., Stathopouios E.N., Stournaras C., Gravanis A., Castanas E. 2005.  Membrane androgen receptor activation induces apoptosis regression of human prostate cancer cells in vitro and in vivo. J Clin Endocrinol Metab, 90(2): 893-903.

22- Hengartner M.O. 2000. The biochemistry of opoptosis, Nature, 407: 770-776.

23- Iványi Z., Szabó N., Huber J., Wölfling J., Zupkó I., Szécsi M., Wittmann T., Schneider G. 2012. Synthesis of Dring-substituted (5’R)- and (5’S)-17b-pyrazolinylandrostene epimers and comparison of their potential anticancer activities. Steroids, 77: 566-574.

24- Jakimov D.S., Kojic V.V., Aleksic L.D., Bogdanovic G.M., Ajdukovic JJ., Djurendic E.A., Penov-Gaši K.M., Sakač M.N., Jovanović-Šanta S.S. 2015. Androstane derivatives induce apoptotic death in MDA-MB-231 breast cancer cells. Bioorg. Med. Chem. 23: 7189-7198.

25- Jiang X., Wang X. 2004.  Cytochrome C-mediated apoptosis, Ann Rev Biochem, 73: 87-106.

26- Kallergi G., Agelaki S., Markomanolaki H., Georgoulias V., Stournaras C. 2007. Activation of FAK/PI3K/Racl signaling controls actin reorganization and inhibits cell motility in human cancer cells, Cell Physiol Biochem, 20: 977-986.

27- Kampa M., Kogia C., Theodoropoulos PA., Anezinis P., Charalampopoulos I., Papakonstanti E.A., Efstathios N Stathopoulos, Hatzoglou A., Stournaras C., Gravanis A., Castanas E. 2006. Activation of membrane androgen receptors potentiates the antiproliferative effects of paclitaxel on human prostate cancer cells. Mol Cancer Ther, 10.1158/1535-7163.

28. Kampa M., Papakonstanti E.A., Hatzoglou A., Stathopoulos E.N., Stournaras C., Castanas E. 2002. The human prostate cancer cell line LNCaP bears functional membrane testosterone receptors that increase PSA secretion and modify actin cytoskeletone. FASEB J, 16: 1429-1431.

29. Kampa M., Pelekanou V., Castanas E. 2008. Membrance- initiated steroid action in breast and prostate cancer, Steroid. 73(9-10): 953-960.

30- Kaufmann S.H., Earnshaw W.C. 2000. Induction of opoptosis by cancer chemotherapy. Exp Cell Res, 256(1): 42-49.

31- Kim R., Emi M., Tanbe K., Murakami S., Uchida Y., Arihiro K., 2006.  Regulation and interplay of apoptotic and non-apaptotic Cell Death Journal pathol, 208: 319-326.

32- Kischkel F.C., Lawrence D.A., Tinel A., LeBlan H., Virmani A., Schow P., Gazdar A., Blenis J., Arnott D., Ashkenazi A. 2001.  Death receotor recruitment of caspase-8, Journal Bid chem, Dec, Epub, 276(49): 46639-46646.

33 - Kuhn C.M. 2002. anabolic steroids. Recent Progress in Hormone Research. The Yale journal of biology and medicine, 57(1):411-434.

34 -  Kumar S., Lavin M.F. 1996. The ICE family of cysteine proteases as effectors of cell death, Cell Death Differ, 3: 255-267.

35-  Kofler R., Schmidt S., Kofler A., and Ausserlechner M. J. 2003. Resistance to glucocorticoid-induced apoptosis in lymphoblastic leukemia. J. Endocrinol, 178(1): 19-27.

36- Kwon A.M., Magnuson B.A. 2007.  Aging alters acute apopotic response to azoxymethane in the colon of rats, Exp Gerontol , 142: 1154-1161.

37- Liamarkopoulos E., Gazouli M., Aravantinos G., Tzanakis N., Theodoropoulos G., Rizos S., Nikiteas N. 2011. Caspase 8 and caspase 9 gene polymorphisms and susceptibility to gastric cancer. Gastric Cancer. 14: 317-321.

38- Li P., Nijhawan D., Budihardjo I., Srinivasula SM., Ahmad M., Alnemri E.S., Wang X. 1997. Cytochrome c and dATP depen-dent formation of Apaf-1/Caspase-9 complex initiates an apoptoticprotease cascade. Cell, 91: 479-489.

39- Li H., Zhu H., Xu C.J., Yuan J. 1998.  Cleavage of BID by Caspase 8 Mediates the Mitochondrial Damage in the Fas Pathway of Apoptosis Cell, 94: 491–501.

40- Morley J.E., Kaiser F.E., Perry H.M., Patrick P., Morley P.M., Stauber P.M., B Vellas, Baumgartner R.N., Baumgartner R.N., Garry P.J. 1997. Longitudinal changes in testosterone, luteinizing hormone, and follicle-stimulating hormone in healthy older men. Metabolism, 46: 410-413.

41- Papadopoulou N., Charalampopoulos I., Anagnostopoulou V., Konstantinidis G., Föller M., Gravanis A., Alevizopoulos K., Lang F., Stournaras C. 2008.  Membrane androgen receptor activation triggers down-regulation of PI-3k/Akt/NF-κB activity and induces apoptotic responses via Bad, FasL and caspase-3 in DU145 prostate cancer cells. Mol. Cancer, 7: 88.

42- Papadopoulo N., Papakonstanti EA., Kallergi G., Alevizopoulos K., Stournaras C. 2009. Membrane androgen receptor activation in prosate and breast tumor cells: Molecular signaling and clinical impact. IUBMB Life, 61(1): 56-61.

43- Passeri M., Pedrazzoni M., Pioli G., Butturini L., Ruys A.H., Cortenraad M.G. 1993.  Effects of nandrolone decanoate on bone mass in established osteoporosis. Maturitas. 17(3): 211 -219.

44-  Przemyslaw W., Miroslawa B., Ewa W., Michal D., Marek S., Robert EH. 1997. Molecular study of sex steroid receptor gene expression in human colon and in colorectal carcinomas. Journal of surgical oncology, 64: 3-11.

45- Schneider G., Kiss A., Mernyák E., Benke Z., Wölfling J., Frank É., Noémi Bózsity N.,  Gyovai A., Minorics R., Zupkó I. 2016. Stereo controlled synthesis of the four 16-hydroxymethyl-19-nortestosterone isomers and their antiproliferative activities. Steroids, 105: 113–120.

46- Simmang CL., Senatore P., Lowry A., Hicks T., Burnstein M., Dentsman F., et al. 1999. Practice parameters for detection of colorectal neoplasms. The Standards Committee, the American Society of Colon and Rectal Surgeons. Dis Colon Rectum, 42(9):1123-9. 

47- Simoes B.M., Alferez D.G., Howell S.J., Clarke RB. 2015. The role of steroid hormones in breast cancer stem cells. Endocr Relat Cancer, 22(6): T177–T186.

48- Slattery M.L., Sweeney C., Murtaugh M., Ma K.N., Wolff R.K., Potter J.D., Caan B.J., Samowitz w. 2005. Associations between ERalpha, ERbeta, and AR genotypes and colon and rectal cancer. Cancer Epidemiol Biomark Prev, 14(12):2936–42.

49- Slee E.A., Harte M.T., Kluck R.M., Wolf B.B., Casiano C.D., Newmeyer DD., Wang H.G.,  Reed JC.,  Nicholson DW., Alnemri E.S., Green D.R.,  Martin S.J. 1999. Ordering the cytochrome c-initiated caspase cascade: hierarchical activation of caspase-2, -3, -6, -7, -8, and -10 in a caspase-9- dependent manner. Journal Cell Biol, 144:281-92.

50- Smith R.A, Eschenbach A.c., Wender R., Levin B., Byers T., Rothenberg D., Brooks D., Creasman W., Cohen C., Runowicz C., Saslow D., Cokkinides V., Eyre H. 2001. American Cancer Society guidelines for the early detection of cancer: Update of early detection guid-lines for prostate, colorectal and Endometrial cancers. Also: update testing for early lung cancer detection. CA Cancer detection, 51:38129-75.

51- Somjen D., Kohen F., Gayer B., Kulik T., Knoll E., Stern N. 2004. Role of putative membrane receptors in the effect of androgen on human vascular cell growth. J Endocrinol, 180: 97- 106.

52 - Srinivasula S.M., Amad M., Fenande S., Alnemri T., Alnemri E.S. 1998.  autoactivationof procaspase-9 by apaf-1-mediated oligomerization. Mol Cell, 1(7), 949-57.

53- Tuckermann J.P., Kleiman A., McPherson K.G., Reichardt H.M. 2005. Molecular mechanisms of glucocorticoids in the control of inflammation and lymphocyte apoptosis. Crit. Rev. Clin. Lab. Sci, 42: 71–104.

54- Valadez-Comes P., German-Castelan, L., Gonzalez A., Velasco-Velazquez M.A., Hansberg-PAstore V., Camacho-Arroyo I. 2015.  Expression and hormonal regulation of membrane progesterone receptors in human astrocytoma cells. Journal Steroid Biochem Mol Biom, 154: 176-85.

55- Virigil T.G., John T.B., Jennifer N.D, Andre M.K., Alan G.P., Patricia V.S.   2004. Mifepristone induces growth arrest, caspase activation, and apoptosis of estrogen receptor-expressing, antiestrogen-resistant breast cancer cells.Clin Cancer Res, 10(15):5215-25.

56 - Wickhom p., Lassere Y. 2007. editors The ABCs of colorectal Cancer.  Seminors in oncology nursing,  Elsevier.

57- Winawer S.J., Fletcher R.H., Miller L., Godlee F., Stolar M.H., Mulrow C.D., S H Woolf S.H., Glick S.N., Ganiats TG., Bond J.M., Rosen L., Zapka J.Z., Olsen S.J., Giardiello FM., Sisk J.E., Antwerp R.V., C Brown-Davis C., Marciniak D.A., Mayer RJ. 1997. Colorectal cancer screening: Clinical guidelines and rationale. Gastroenterology, 112(2), 594-642.

58-  Wobbes T.H., Beex L.V.A.M., Koenders A.M.J. 1984. Estrogen and progestin receptors in Colonic cancer. Dis colon rectu, 35, 163- 165.