Biopolym. Cell. 2014; 30(1):54-60.
Molecular Biomedicine
Effect of anticancer drugs on breast cancer cells sensitive and resistant to doxorubicin: expression of mRNAs of TGF-β and its receptors
1Chorna I. V., 2Fedorenko O. V., 2Stoika R. S.
  1. Sumy State University
    2, Rymskogo-Korsakova., Sumy, Ukraine, 40007
  2. Institute of Cell Biology, NAS of Ukraine
    14/16, Drahomanov Str., Lviv, Ukraine, 79005


Aim. Comparative study of the effect of chemotherapeutic drugs (doxorubicin, methotrexate and cisplatin) and TGF-β on the human breast carcinoma MCF-7 cells, sensitive (wt) and resistant (DOX/R) to the doxorubicin action. Methods. Semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) was used for the estimation of expression of mRNAs coding for the TGF-β isoforms (TGF-β1 and TGF-β2) and the TGF-β type I and II receptors (TRI and TRII). Trypan blue exclusion method was used for measuring cell number and cell viability. Results. The MCF-7(DOX/R) cells were more refractory to the TGF1-dependent growth inhibition than the MCF-7(wt) cells. The level of mRNAs coding for TGF and its receptors was higher in the untreated MCF-7 (DOX/R) cells comparing to the MCF-7(wt) cells. The expression of mRNA coding for TRII was decreased in both cell lines treated with doxorubicin, methotrexate and cisplatin, while the down-regulation of mRNA coding for TRI was revealed only in the MCF-7(DOX/R) cells upon the treatment with doxorubicin and methotrexate. Conclusions. The differential effects of studied anticancer drugs and TGF-β on the doxorubicin-sensitive and -resistant cells have been demonstrated. The elucidation of the molecular mechanisms of escape of the MCF-7 (DOX/R) cells from the growth inhibition by TGF-β requires further investigation.
Keywords: MCF-7, TGF-, doxorubicin, methotrexate, cisplatin, resistance


[1] Aljarrah K, Mhaidat NM, Al-Akhras MA, Aldaher AN, Albiss B, Aledealat K, Alsheyab FM. Magnetic nanoparticles sensitize MCF-7 breast cancer cells to doxorubicin-induced apoptosis. World J Surg Oncol. 2012; 10:62.
[2] Minotti G, Menna P, Salvatorelli E, Cairo G, Gianni L. Anthracyclines: molecular advances and pharmacologic developments in antitumor activity and cardiotoxicity. Pharmacol Rev. 2004; 56(2):185–229.
[3] Florea AM, Busselberg D. Cisplatin as an anti-tumor drug: cellular mechanisms of activity, drug resistance and induced side effects. Cancers (Basel). 2011; 3(1):1351–71.
[4] Skubisz MM, Tong S. The evolution of methotrexate as a treatment for ectopic pregnancy and gestational trophoblastic neoplasia: a review. ISRN Obstet Gynecol. 2012; 2012:637094.
[5] Baguley BC. Multiple drug resistance mechanisms in cancer. Mol Biotechnol. 2010; 46(3):308–16.
[6] Gillet JP, Gottesman MM. Mechanisms of multidrug resistance in cancer. Methods Mol Biol. 2010; 596:47–76.
[7] Higgins CF. Multiple molecular mechanisms for multidrug resistance transporters. Nature. 2007; 446(7137):749–57.
[8] Chekhun VF, Kulik GI, Yurchenko OV, Tryndyak VP, Todor IN, Luniv LS, Tregubova NA, Pryzimirska TV, Montgomery B, Rusetskaya NV, Pogribny IP. Role of DNA hypomethylation in the development of the resistance to doxorubicin in human MCF-7 breast adenocarcinoma cells. Cancer Lett. 2006; 231(1):87–93.
[9] Lukyanova NY, Rusetskya NV, Tregubova NA, Chekhun VF. Molecular profile and cell cycle in MCF-7 cells resistant to cisplatin and doxorubicin. Exp Oncol. 2009; 31(2):87–91.
[10] Connolly EC, Freimuth J, Akhurst RJ. Complexities of TGF-b targeted cancer therapy. Int J Biol Sci. 2012; 8(7):964–78.
[11] Papadopoulou E, Anagnostopoulos K, Tripsianis G, Tentes I, Kakolyris S, Galazios G, Sivridis E, Simopoulos K, Kortsaris A. Evaluation of predictive and prognostic significance of serum TGF-beta1 levels in breast cancer according to HER-2 codon 655 polymorphism. Neoplasma. 2008; 55(3):229–38.
[12] Dancea HC, Shareef MM, Ahmed MM. Role of radiation-induced TGF-beta signaling in cancer therapy. Mol Cell. Pharmacol. 2009; 1(1):44–56.
[13] Barcellos-Hoff MH, Akhurst RJ. Transforming growth factorbeta in breast cancer: too much, too late. Breast Cancer Res. 2009; 11(1):202.
[14] Calone I, Souchelnytskyi S. Inhibition of TGFb signaling and its implications in anticancer treatments. Exp Oncol. 2012; 34(1): 9–16.
[15] Chowdhury S, Ammanamanchi S, Howell GM. Epigenetic targeting of transforming growth factor b receptor II and implications for cancer therapy. Mol Cell Pharmacol. 2009; 1(1):57–70.
[16] Chorna I, Fedorenko O, Datsyuk L, Stoika R. Expression of mRNA coding for TGF-beta and its receptors in irradiated human breast carcinoma MCF-7 cells differing in their sensitivity to doxorubicin. Exp Oncol. 2005; 27(2):156–8.
[17] Filyak O, Stoika RS. Comparative study of p53 expression in human carcinoma cell lines A549 and MCF7 under anticancer drug treatment. Ukr Biokhim Zh. 2005; 77(2):136–40.
[18] Chorna I, Bilyy R, Datsyuk L, Stoika R. Comparative study of human breast carcinoma MCF-7 cells differing in their resistance to doxorubicin: effect of ionizing radiation on apoptosis and TGFb production. Exp Oncol. 2004; 26(2):111–7.
[19] Chorna IV, Datsyuk LO, Stoika RS. Expression of Bax, Bad and Bcl-2 proteins under X-radiation effect towards human breast carcinoma MCF-7 cells and their doxorubicin-resistant derivatives. Exp Oncol. 2005; 27(3):196–201.
[20] Stoika RS, Yakymovych IA, Kashchak NI, Boyko MM, Korynevska AV, Klyuchyvska OY, Shafranska GI, Yakymovych MY, Zhylchuk VY, Kudryavets YY, Vorontsova AL. Effect of anticancer drugs on production of transforming growth factor and expression of p53 AND Bcl-2 proteins by MCF-7 and T47D cell lines of human breast carcinoma. Exp Oncol. 2008; 30(1):35–41.
[21] Sheen-Chen SM, Chen HS, Sheen CW, Eng HL, Chen WJ. Serum levels of transforming growth factor beta1 in patients with breast cancer. Arch Surg. 2001; 136(8):937–40.
[22] Gonzalez-Santiago AE, Mendoza-Topete LA, Sanchez-Llamas F, Troyo-Sanroman R, Gurrola-Diaz CM. TGF-b1 serum concentration as a complementary diagnostic biomarker of lung cancer: establishment of a cut-point value. J Clin Lab Anal. 2011; 25 (4):238–43.
[23] Kemik O, Kemik AS, Purisa S, Hasirci I, Dulger AC, Adas M, Tuzun S. Transforming growth factor beta-1 in human colorectal cancer patients. Eur J Gen Med. 2011; 8(1):53–6.
[24] Shim KS, Kim KH, Han WS, Park EB. Elevated serum levels of transforming growth factor-b1 in patients with colorectal carcinoma: its association with tumor progression and its significant decrease after curative surgical resection. Cancer. 1999; 85(3): 554–61.
[25] Adler HL, McCurdy MA, Kattan MW, Timme TL, Scardino PT, Thompson TC. Elevated levels of circulating interleukin-6 and transforming growth factor-beta1 in patients with metastatic prostatic carcinoma. J Urol. 1999; 161(1):182–7.
[26] Jaowlew SB. Transforming growth factor-beta in cancer and metastasis. Cancer Metastasis Rev. 2006; 25(3):435–57.
[27] Stoika R, Yakymovych M, Souchelnytskyi S, Yakymovych I. Potential role of transforming growth factor beta1 in drug resistance of tumor cells. Acta Biochim Pol. 2003; 50(2):497–508.
[28] Gomes LR, Terra LF, Wailemann RA, Labriola L, Sogayar MC. TGF-b1 modulates the homeostasis between MMPs and MMP inhibitors through p38 MAPK and ERK1/2 in highly invasive breast cancer cells. BMC Cancer. 2012; 12:26.
[29] Nagaraj NS, Datta PK. Targeting the transforming growth factor-beta signaling pathway in human cancer. Expert Opin Investig Drugs. 2010; 19(1):77–91.
[30] Biswas S, Trobridge P, Romero-Gallo J, Billheimer D, Myeroff LL, Willson JK, Markowitz SD, Grady WM. Mutational inactivation of TGFBR2 in microsatellite unstable colon cancer arises from the cooperation of genomic instability and the clonal outgrowth of transforming growth factor beta resistant cells. Genes Chromosomes Cancer. 2008; 47(2):95–106.
[31] Kim SJ, Im YH, Markowitz SD, Bang YJ. Molecular mechanisms of inactivation of TGF-beta receptors during carcinogenesis. Cytokine Growth Factor Rev. 2000; 11(1–2):159–68.
[32] Fujiwara K, Ikeda H, Yoshimoto T. Abnormalities in expression of genes, mRNA, and proteins of transforming growth factorbeta receptor type I and type II in humanpituitary adenomas. Clin Neuropathol. 1998; 17(1):19–26.
[33] Izumoto S, Arita N, Ohnishi T, Hiraga S, Taki T, Tomita N, Ohue M, Hayakawa T. Microsatellite instability and mutated type II transforming growth factor-beta receptor gene in gliomas. Cancer Lett. 1997; 112(2):251–6.
[34] Kubiczkova L, Sedlarikova L, Hajek R, Sevcikova S. TGF-b – an excellent servant but a bad master. J Transl Med. 2012; 10:183.
[35] Kellman C, Olofsson SP, Hansson O, Von Schantz T, Lindvall M, Nilsson I, Salford LG, Sjogren HO, Widegren B. Expression of TGF-b isoforms, TGF-b receptors, and Smad molecules at different stages of human glioma. Int J Cancer. 2000; 89(3):251–8.
[36] Yamada N, Kato M, Yamashita H, Nister M, Miyazono K, Heldin CH, Funa K. Enhanced expression of transforming growth factor-b and its type-I and type-II receptors in human glioblastoma. Int J Cancer. 1995; 62(4):386–92.
[37] Filyak Y, Filyak O, Stoika R. Transforming growth factor beta-1 enhances cytotoxic effect of doxorubicin in human lung adenocarcinoma cells of A549 line. Cell Biol Int. 2007; 31(8):851–5.