Increased chemoresistance to paclitaxel in the MCF10AT series of human breast epithelial cancer cells

Lim,Soo-Jeong; Choi,Hyeon Gyeom; Jeon,Chae Kyung; Kim,So Hee

doi:10.3892/or.2015.3775

Increased chemoresistance to paclitaxel in the MCF10AT series of human breast epithelial cancer cells

Authors:
- Soo-Jeong Lim
- Hyeon Gyeom Choi
- Chae Kyung Jeon
- So Hee Kim
View Affiliations

Affiliations: Department of Bioscience and Biotechnology, Sejong University, Seoul, Republic of Korea, College of Natural Science, Hannam University, Daejeon, Republic of Korea, Grinnell College, Grinnell, IA, USA, College of Pharmacy and Research Institute of Pharmaceutical Science and Technology, Ajou University, Suwon, Republic of Korea
Published online on: February 2, 2015 https://doi.org/10.3892/or.2015.3775
Pages: 2023-2030

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The MCF10AT cell series of human breast epithelial cancer cells includes normal MCF10A (10A), premalignant MCF10AT (10AT) and MCF10ATG3B (10ATG3B), and fully malignant MCF10CA1a (10CA1a) cells. The series is a unique model system showing progressive tumorigenic potential with the same origin. The effects of paclitaxel, a microtubule inhibitor, were evaluated in this cell system. Paclitaxel inhibited cell proliferation in a time-dependent (24, 48 and 72 h) and concentration-dependent (0-10 nM) manners with less sensitivity in 10CA1a cells. Treatment with paclitaxel (10 nM) for 24 h induced apoptosis in 10A, 10AT, 10ATG3B and 10CA1a cells, with 23.6, 26.1, 25.2 and 8.96%, respectively, in the sub-G1 phase. Treatment with paclitaxel (0-10 nM) for 24 h, resulted in the appearance of DNA fragmentation (a hallmark of apoptosis) with less sensitivity in the 10CA1a tumor cells. Paclitaxel increased p53 protein expression in 10A, 10AT, 10ATG3B and 10CA1a cells, by 87, 102, 812 and 84%, respectively. The p21Waf1/Cip1 protein expression increased by 2.57-, 1.53- and 2.48-fold in 10A, 10AT and 10ATG3B cells, respectively, with negligible detection in the 10CA1a cells. Activation of the Akt signaling pathway was observed in the MCF10AT cell lineage and the protein expression of phospho-Akt (Ser473 and Thr308). The downstream targets of this pathway, phospho-p70S6K and phospho-S6RP, were also inhibited by paclitaxel in 10A, 10AT and 10ATG3B cells, but minimally inhibited in 10CA1a cells, suggestive of chemoresistance in 10CA1a cells. The effects of paclitaxel on the multidrug resistance 1 (MDR1), MRP1 and breast cancer resistance protein (BCRP) gene expression were not significant in the MCF10AT cell lineage. These results collectively indicated that paclitaxel inhibited cell proliferation and induced apoptosis in the MCF10AT cell lineage, with chemoresistance in 10CA1a tumor cells. The decreased responsiveness to paclitaxel observed in 10CA1a tumor cells was likely due, in part, to activation of the Akt signaling pathway and a high expression of wild-type p53 with lack of p21Waf1/Cip1.

View Figures

View References

1	Jemal A, Tiwari RC, Murray T, Ghafoor A, Samuels A, Ward E, Feuer EJ and Thun MJ: Cancer statistics. CA Cancer J Clin. 54:8–29. 2004. View Article : Google Scholar : PubMed/NCBI
2	Dawson PJ, Wolman SR, Tait L, Heppner GH and Miller FR: MCF10AT: a model for the evolution of cancer from proliferative breast disease. Am J Pathol. 148:313–319. 1996.PubMed/NCBI
3	Soule HD, Maloney TM, Wolman SR, Peterson WD Jr, Brenz R, McGrath CM, Russo J, Pauley RJ, Jones RF and Brooks SC: Isolation and characterization of a spontaneously immortalized human breast epithelial cell line, MCF-10. Cancer Res. 50:6075–6086. 1990.PubMed/NCBI
4	Kim SH, Miller FR, Tait L, Zheng J and Novak RF: Proteomic and phosphoproteomic alterations in benign, premalignant and tumor human breast epithelial cells and xenograft lesions: biomarkers of progression. Int J Cancer. 124:2813–2828. 2009. View Article : Google Scholar : PubMed/NCBI
5	Kim SH, Zukowski K and Novak RF: Rapamycin effects on mTOR signaling in benign, premalignant and malignant human breast epithelial cells. Anticancer Res. 29:1143–1150. 2009.PubMed/NCBI
6	Jordan MA: Mechanism of action of antitumor drugs that interact with microtubules and tubulin. Curr Med Chem Anticancer Agents. 2:1–17. 2002. View Article : Google Scholar
7	Guéritte F: General and recent aspects of the chemistry and structure-activity relationships of taxoids. Curr Pharm Des. 7:1229–1249. 2001. View Article : Google Scholar : PubMed/NCBI
8	Ramanathan B, Jan KY, Chen CH, Hour TC, Yu HJ and Pu YS: Resistance to paclitaxel is proportional to cellular total antioxidant capacity. Cancer Res. 65:8455–8460. 2005. View Article : Google Scholar : PubMed/NCBI
9	Fuchs DA and Johnson RK: Cytologic evidence that taxol, an antineoplastic agent from Taxus brevifolia, acts as a mitotic spindle poison. Cancer Treat Rep. 62:1819–1222. 1978.
10	Lin HL, Liu TH, Chau GY, Lui WY and Chi CW: Comparison of 2-methoxyestradiol-induced, docetaxel-induced, and paclitaxel-induced apoptosis in hepatoma cells and its correlation with reactive oxygen species. Cancer. 89:983–994. 2000. View Article : Google Scholar : PubMed/NCBI
11	Dziadyk JM, Sui M, Zhu X and Fan W: Paclitaxel-induced apoptosis may occur without a prior G₂/M-phase arrest. Anticancer Res. 24:27–36. 2004.PubMed/NCBI
12	Zhou J and Giannakakou P: Targeting microtubules for cancer chemotherapy. Curr Med Chem Anticancer Agents. 5:65–71. 2005. View Article : Google Scholar : PubMed/NCBI
13	McGrogen BT, Gilmartin B, Carney DN and McCann A: Taxanes, microtubules and chemoresistant breast cancer. Biochim Biohys Acta. 1785:96–132. 2008.
14	Villeneuve DJ, Hembruff SL, Veitch Z, Cecchetto M, Dew WA and Parissenti AM: cDNA microarray analysis of isogenic paclitaxel- and doxorubicin-resistant breast tumor cell lines reveals distinct drug-specific genetic signatures of resistance. Breast Cancer Res Treat. 96:17–39. 2006. View Article : Google Scholar
15	Starcevic SL, Elferink C and Novak RF: Progressive resistance to apoptosis in a cell lineage model of human proliferative breast disease. J Natl Cancer Inst. 93:776–782. 2001. View Article : Google Scholar : PubMed/NCBI
16	Starcevic SL, Diotte NM, Zukowski KL, Cameron MJ and Novak RF: Oxidative DNA damage and repair in a cell lineage model of human proliferative breast disease (PBD). Toxicol Sci. 75:74–81. 2003. View Article : Google Scholar : PubMed/NCBI
17	Basolo F, Elliott J, Tait L, Chen XQ, Maloney T, Russo IH, Pauley R, Momiki S, Caamano J, Klein-Szanto AJ, et al: Transformation of human breast epithelial cells by c-Ha-ras oncogene. Mol Carcinog. 4:25–35. 1991. View Article : Google Scholar : PubMed/NCBI
18	Russo J, Tait L and Russo IH: Morphological expression of cell transformation induced by c-Ha-ras oncogene in human breast epithelial cells. J Cell Sci. 99:453–463. 1991.PubMed/NCBI
19	Santner SJ, Dawson PJ, Tait L, Soule HD, Eliason J, Mohamed AN, Wolman SR, Heppner GH and Miller F: Malignant MCF10CA1 cell lines derived from premalignant human breast epithelial MCF10AT cells. Breast Cancer Res Treat. 65:101–110. 2001. View Article : Google Scholar : PubMed/NCBI
20	Liu S, Wu D, Li L, Sun X, Xie W and Li X: NF-κB activation was involved in reactive oxygen species-mediated apoptosis and autophagy in 1-oxoeudesm-11(13)-eno-12,8α-lactone-treated human lung cancer cells. Arch Pharm Res. 37:1039–1052. 2014. View Article : Google Scholar
21	Zhan Y, Chen Y, Liu R, Zhang H and Zhang Y: Potentiation of paclitaxel activity by curcumin in human breast cancer cell by modulating apoptosis and inhibiting EGFR signaling. Arch Pharm Res. 37:1086–1095. 2014. View Article : Google Scholar
22	Lee EJ, Park MK, Kim HJ, Kang JH, Kim YR, Kang GJ, Byun HJ and Lee CH: 12-O-Tetradecanoylphorbol-13-acetate induces keratin 8 phosphorylation and reorganization via expression of transglutaminases-2. Biomol Ther. 22:122–128. 2014. View Article : Google Scholar
23	Zhang L, Wei Y, Pushel I, Heinze K, Elenbaas J, Henry RW and Arnosti DN: Integrated stability and activity control of the Drosophila Rbf1 retinoblastoma protein. J Biol Chem. 289:24863–24873. 2014. View Article : Google Scholar : PubMed/NCBI
24	Fresno Vara JA, Casado E, de Castro J, Cejas P, Belda-Iniesta C and González-Barón M: PI3K/Akt signaling pathway and cancer. Cancer Treat Rev. 30:193–204. 2004. View Article : Google Scholar : PubMed/NCBI
25	Kong HJ, Yu HJ, Hong S, Park MJ, Choi YH, An WG, Lee JW and Cheong J: Interaction and functional cooperation of the cancer-amplified transcriptional coactivator activating signal cointegrator-2 and E2F-1 in cell proliferation. Mol Cancer Res. 1:948–958. 2003.PubMed/NCBI
26	Kim CW, Lu JN, Go SI, Jung JH, Yi SM, Jeong JH, Hah YS, Han MS, Park JW, Lee WS and Min YJ: p53 restoration can overcome cisplatin resistance through inhibition of Akt as well as induction of Bax. Int J Oncol. 43:1495–1502. 2013.PubMed/NCBI
27	Sezgin C, Karabulut B, Uslu R, Sanli UA, Goksel G, Zekioglu O, Ozdemir N and Goker E: Potential predictive factors for response to weekly paclitaxel treatment in patients with metastatic breast cancer. J Chemother. 17:96–103. 2005. View Article : Google Scholar : PubMed/NCBI
28	Schmidt M, Bachhuber A, Victor A, Steiner E, Mahlke M, Lehr HA, Pilch H, Weikel W and Knapstein P: p53 expression and resistance against paclitaxel in patients with metastatic breast cancer. J Cancer Res Clin Oncol. 129:295–302. 2003.PubMed/NCBI
29	Waldman T, Kinzler KW and Vogelstein B: p21 is necessary for the p53-mediated G₁ arrest in human cancer cells. Cancer Res. 55:5187–5190. 1995.PubMed/NCBI
30	Knuefermann C, Lu Y, Liu B, Jin W, Liang K, Wu L, Schmidt M, Mills GB, Mendelsohn J and Fan Z: HER2/PI-3K/Akt activation leads to a multidrug resistance in human breast adenocarcinoma cells. Oncogene. 22:3205–3212. 2003. View Article : Google Scholar : PubMed/NCBI
31	Clark AS, West K, Streicher S and Dennis PA: Constitutive and inducible Akt activity promotes resistance to chemotherapy, trastuzumab, or tamoxifen in breast cancer cells. Mol Cancer Ther. 1:707–717. 2002.PubMed/NCBI
32	Chang F, Lee JT, Navolanic PM, Steelman LS, Shelton JG, Blalock WL, Franklin RA and McCubrey JA: Involvement of the P13K/Akt pathway in cell cycle progression, apoptosis, and neoplastic transformation: a target for cancer chemotherapy. Leukemia. 17:590–603. 2003. View Article : Google Scholar : PubMed/NCBI
33	Page C, Lin HJ, Jin Y, Castle VP, Nunez G, Huang M and Lin J: Overexpression of Akt/AKT can modulate chemotherapy-induced apoptosis. Anticancer Res. 20:407–416. 2000.PubMed/NCBI
34	Hu L, Hofmann J, Lu Y, Mills GB and Jaffe RB: Inhibition of phosphatidylinositol 3′-kinase increases efficacy of paclitaxel in in vitro and in vivo ovarian cancer models. Cancer Res. 62:1087–1092. 2002.PubMed/NCBI
35	Shingu T, Yamada K, Hara N, Moritake K, Osago H, Terashima M, Uemura T, Yamasaki T and Tsuchiya M: Synergistic augmentation of antimicrotubule agent-induced cytotoxicity by a phosphoinositide 3-kinase inhibitor in human malignant glioma cells. Cancer Res. 63:4044–4047. 2003.PubMed/NCBI
36	Nguyen DM, Chen GA, Reddy R, Tsai W, Schrump WD, Cole G Jr and Schrump DS: Potentiation of paclitaxel cytotoxicity in lung and esophageal cancer cells by pharmacologic inhibition of the phosphoinositide 3-kinase/protein kinase B (Akt)-mediated signaling pathway. J Thorac Cardiovasc Surg. 127:365–375. 2004. View Article : Google Scholar : PubMed/NCBI
37	Cheng JQ, Lindsley CW, Cheng GZ, Yang H and Nicosia SV: The Akt/PKB pathway: molecular target for cancer drug discovery. Oncogene. 24:7482–7492. 2005. View Article : Google Scholar : PubMed/NCBI