[Frontiers in Bioscience, Landmark, 25, 948-960, March 1, 2020]

PCAT1/miR-129/ABCB1 axis confers chemoresistance in non-small cell lung cancer

Ruifen Tian1, Congjun Zhang2, Fuxing Xiong2, Huanwen Chen3

1Department of respiration, Shanxi Tumour Hospital, Taiyuan 030013, Shanxi Province, China, 2Medical Oncology, the First Affiliated Hospital of Anhui Medical University; Hefei 230022, Anhui Province, China, 3Cardiothoracic surgery, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China

TABLE OF CONTENTS

1. Abstract
2. Introduction
3. Materials and methods
    3.1. Patients and tumor tissues
    3.2. Cell culture
    3.3. Cell transfection
    3.4. Detection of cell proliferation capacity
    3.5. Cell apoptosis assay
    3.6. Luciferase reporter assay
    3.7. Western blot analysis
    3.8. Quantitative real-time polymerase chain reaction (qRT-PCR)
    3.9. Statistical analysis
4. Results
    4.1. lncRNA PCAT1 is upregulated in NSCLC tissues and cell lines
    4.2. Knockdown of PCAT1 improves the sensitivity of NSCLC cells to DDP
    4.3. PCAT1 acts as a ‘sponge’ for miR-129 in NSCLC cells
    4.4. ABCB1 upregulation by the PCAT1/miR-129 axis contributes to DDP resistance in DDP-resistant NSCLC cells
5. Discussion
6. Acknowledgments
7. References

1. ABSTRACT

Long noncoding RNA prostate cancer-associated transcript 1 (PCAT1) is oncogenic and causes progression of non-small cell lung cancer (NSCLC). We hypothesized that PCAT1 might be involved in the acquisition of chemoresistance of NSCLC cells to treatment with cisplatin (DDP). Here, we show that PCAT1 and ATP-binding cassette sub-family B member 1 (ABCB1) are highly expressed in NSCLC tissues and cell lines, and regulate the growth and apoptosis of these cells. Compared with those in DDP-sensitive patients, PCAT1 and ABCB1 are highly expressed in the tumors of DDP-resistant patients, and such overexpression correlates with a shorter overall survival of these patients. Knockdown of PCAT1 or upregulation of miR-129 led to apoptosis and sensitized the DDP-resistant cells to DDP. The 3’ UTR activity of PCAT1 and ABCB1, which was increased by PCAT1 overexpression, was shown to harbor an miR-129 binding site. DDP resistance is induced by elevated ABCB1 expression, which involves binding of miR-129 in DDP resistant cells. These findings suggest that the PCAT1/miR-129/ABCB1 axis may be a potential target for the treatment of DDP-resistant oat cell cancer.

7. REFERENCES

1. L. A. Torre, F. Bray, R. L. Siegel, J. Ferlay, J. Lortet-Tieulent and A. Jemal: Global cancer statistics, 2012. CA Cancer J Clin, 65(2), 87-108 (2015)
DOI: 10.3322/caac.21262

2. J. R. Molina, P. Yang, S. D. Cassivi, S. E. Schild and A. A. Adjei: Non-small cell lung cancer: epidemiology, risk factors, treatment, and survivorship. Mayo Clin Proc, 83(5), 584-94 (2008)
DOI: 10.4065/83.5.584

3. T. Okamoto, Y. Suzuki, T. Fujishita, H. Kitahara, S. Shimamatsu, M. Kohno, Y. Morodomi, D. Kawano and Y. Maehara: The prognostic impact of the amount of tobacco smoking in non-small cell lung cancer--differences between adenocarcinoma and squamous cell carcinoma. Lung Cancer, 85(2), 125-30 (2014)
DOI: 10.1016/j.lungcan.2014.06.006

4. H. RS, M. D and B. C: The biology and management of non-small cell lung cancer. Nature, 553(7689), 446-454 (2018)
DOI: 10.1038/nature25183

5. D. A. Fennell, Y. Summers, J. Cadranel, T. Benepal, D. C. Christoph, R. Lal, M. Das, F. Maxwell, C. Visseren-Grul and D. Ferry: Cisplatin in the modern era: The backbone of first-line chemotherapy for non-small cell lung cancer. Cancer Treat Rev, 44, 42-50 (2016)
DOI: 10.1016/j.ctrv.2016.01.003

6. J. H. Bergmann and D. L. Spector: Long non-coding RNAs: modulators of nuclear structure and function. Curr Opin Cell Biol, 26, 10-8 (2014)
DOI: 10.1016/j.ceb.2013.08.005

7. F. A. Karreth and P. P. Pandolfi: ceRNA cross-talk in cancer: when ce-bling rivalries go awry. Cancer Discov, 3(10), 1113-21 (2013)
DOI: 10.1158/2159-8290.CD-13-0202

8. D. Chakravarty, A. Sboner, S. S. Nair, E. Giannopoulou, R. Li, S. Hennig, J. M. Mosquera, J. Pauwels, K. Park, M. Kossai, T. Y. MacDonald, J. Fontugne, N. Erho, I. A. Vergara, M. Ghadessi, E. Davicioni, R. B. Jenkins, N. Palanisamy, Z. Chen, S. Nakagawa, T. Hirose, N. H. Bander, H. Beltran, A. H. Fox, O. Elemento and M. A. Rubin: The oestrogen receptor alpha-regulated lncRNA NEAT1 is a critical modulator of prostate cancer. Nat Commun, 5, 5383 (2014)
DOI: 10.1038/ncomms6383

9. M. D. Huang, W. M. Chen, F. Z. Qi, R. Xia, M. Sun, T. P. Xu, L. Yin, E. B. Zhang, W. De and Y. Q. Shu: Long non-coding RNA ANRIL is upregulated in hepatocellular carcinoma and regulates cell apoptosis by epigenetic silencing of KLF2. J Hematol Oncol, 8, 50 (2015)
DOI: 10.1186/s13045-015-0146-0

10. X. Wang, M. Li, Z. Wang, S. Han, X. Tang, Y. Ge, L. Zhou, C. Zhou, Q. Yuan and M. Yang: Silencing of Long Noncoding RNA MALAT1 by miR-101 and miR-217 Inhibits Proliferation, Migration, and Invasion of Esophageal Squamous Cell Carcinoma Cells. J Biol Chem, 290(7), 3925 (2015)
DOI: 10.1074/jbc.M114.596866

11. Q. Yuan, H. Chu, Y. Ge, G. Ma, M. Du, M. Wang, Z. Zhang and W. Zhang: LncRNA PCAT1 and its genetic variant rs1902432 are associated with prostate cancer risk. J Cancer, 9(8), 1414-1420 (2018)
DOI: 10.7150/jca.23685

12. X. Zhang, Y. Zhang, Y. Mao and X. Ma: The lncRNA PCAT1 is correlated with poor prognosis and promotes cell proliferation, invasion, migration and EMT in osteosarcoma. Onco Targets Ther, 11, 629-638 (2018)
DOI: 10.2147/OTT.S152063

13. Q. Zhen, L. N. Gao, R. F. Wang, W. W. Chu, Y. X. Zhang, X. J. Zhao, B. L. Lv and J. B. Liu: LncRNA PCAT-1 promotes tumour growth and chemoresistance of oesophageal cancer to cisplatin. Cell Biochem Funct, 36(1), 27-33 (2018)
DOI: 10.1002/cbf.3314

14. H. X and Z. H: Long non-coding RNA PCAT-1 over-expression promotes proliferation and metastasis in non-small cell lung cancer cells.%A Zhao B. Int J Clin Exp Med, 8(10), 18482-7 (2015) (doi not found)

15. G. Di Leva, M. Garofalo and C. M. Croce: MicroRNAs in cancer. Annu Rev Pathol, 9, 287-314 (2014)
DOI: 10.1146/annurev-pathol-012513-104715

16. D. W. Thomson and M. E. Dinger: Endogenous microRNA sponges: evidence and controversy. Nat Rev Genet, 17(5), 272-83 (2016)
DOI: 10.1038/nrg.2016.20

17. T. Kawamata and Y. Tomari: Making RISC. Trends Biochem Sci, 35(7), 368-76 (2010)
DOI: 10.1016/j.tibs.2010.03.009

18. R. M. Drayton, E. Dudziec, S. Peter, S. Bertz, A. Hartmann, H. E. Bryant and J. W. Catto: Reduced expression of miRNA-27a modulates cisplatin resistance in bladder cancer by targeting the cystine/glutamate exchanger SLC7A11. Clin Cancer Res, 20(7), 1990-2000 (2014)
DOI: 10.1158/1078-0432.CCR-13-2805

19. C. Liang, Z. Qi, H. Ge, C. Liang, Y. Zhang, Z. Wang, R. Li and J. Guo: Long non-coding RNA PCAT-1 in human cancers: A meta-analysis. Clin Chim Acta, 480, 47-55 (2018)
DOI: 10.1016/j.cca.2018.01.043

20. L. Qiao, X. Liu, Y. Tang, Z. Zhao, J. Zhang and H. Liu: Knockdown of long non-coding RNA prostate cancer-associated ncRNA transcript 1 inhibits multidrug resistance and c-Myc-dependent aggressiveness in colorectal cancer Caco-2 and HT-29 cells. Mol Cell Biochem, 441(1-2), 99-108 (2018)
DOI: 10.1007/s11010-017-3177-8

21. X. Lu, J. Ma, J. Chu, Q. Shao, Y. Zhang, G. Lu, J. Li, X. Huang, W. Li, Y. Li, Y. Ling and T. Zhao: MiR-129-5p Sensitizes the Response of Her-2 Positive Breast Cancer to Trastuzumab by Reducing Rps6. Cell Physiol Biochem, 44(6), 2346-2356 (2017)
DOI: 10.1159/000486122

22. M. Karaayvaz, H. Zhai and J. Ju: miR-129 promotes apoptosis and enhances chemosensitivity to 5-fluorouracil in colorectal cancer. Cell Death Dis, 4, e659 (2013)
DOI: 10.1038/cddis.2013.193

23. Y. Zhang, Y. Wang, Y. Wei, M. Li, S. Yu, M. Ye, H. Zhang, S. Chen, W. Liu and J. Zhang: MiR-129-3p promotes docetaxel resistance of breast cancer cells via CP110 inhibition. Sci Rep, 5, 15424 (2015)
DOI: 10.1038/srep15424

24. A. Chang: Chemotherapy, chemoresistance and the changing treatment landscape for NSCLC. Lung Cancer, 71(1), 3-10 (2011)
DOI: 10.1016/j.lungcan.2010.08.022

25. B. C. Shaffer, J. P. Gillet, C. Patel, M. R. Baer, S. E. Bates and M. M. Gottesman: Drug resistance: still a daunting challenge to the successful treatment of AML. Drug Resist Updat, 15(1-2), 62-9 (2012)
DOI: 10.1016/j.drup.2012.02.001

26. R. Silva, V. Vilas-Boas, H. Carmo, R. J. Dinis-Oliveira, F. Carvalho, M. de Lourdes Bastos and F. Remiao: Modulation of P-glycoprotein efflux pump: induction and activation as a therapeutic strategy. Pharmacol Ther, 149, 1-123 (2015)
DOI: 10.1016/j.pharmthera.2014.11.013

27. S. Vallo, R. Kopp, M. Michaelis, F. Rothweiler, G. Bartsch, M. P. Brandt, K. M. Gust, F. Wezel, R. A. Blaheta, A. Haferkamp and J. Cinatl, Jr.: Resistance to nanoparticle albumin-bound paclitaxel is mediated by ABCB1 in urothelial cancer cells. Oncol Lett, 13(6), 4085-4092 (2017)
DOI: 10.3892/ol.2017.5986

28. A. P. Lombard, C. Liu, C. M. Armstrong, V. Cucchiara, X. Gu, W. Lou, C. P. Evans and A. C. Gao: ABCB1 Mediates Cabazitaxel-Docetaxel Cross-Resistance in Advanced Prostate Cancer. Mol Cancer Ther, 16(10), 2257-2266 (2017)
DOI: 10.1158/1535-7163.MCT-17-0179

29. C. J, Z. Q, Z. L, C. C, L. L, Z. W and L. F: Long noncoding RNA PCAT-1 promotes invasion and metastasis via the miR-129-5p-HMGB1 signaling pathway in hepatocellular carcinoma.%A Zhang D. Biomed pharmacother, 95, 1187-1193 (2017)
DOI: 10.1016/j.biopha.2017.09.045

30. C. J, D. X and H. J: Long non-coding RNA PCAT-1 contributes to tumorigenesis by regulating FSCN1 via miR-145-5p in prostate cancer.%A Xu W. Biomed pharmacother, 95, 1112-1118 (2017)
DOI: 10.1016/j.biopha.2017.09.019

31. W. M, X. Y, L. Z, L. K, K. P, C. Y and J. X: Long noncoding RNA PCAT1 regulates extrahepatic cholangiocarcinoma progression via the Wnt/β-catenin-signaling pathway. Biomed pharmacother, 94, 55-62 (2017)
DOI: 10.1016/j.biopha.2017.07.025

Abbeviations: Long noncoding RNA (lncRNA), prostate cancer-associated transcript 1 (PCAT1), non-small cell lung cancer (NSCLC), microRNAs (miRNAs), ATP-binding cassette sub-family B member 1 (ABCB1), cisplatin (DDP), Lung adenocarcinoma (LUAD), high mobility group box 1 (HMGB1), competitive endogenous RNA (ceRNAs).

Key Words: Chemoresistance, Non-Small Cell Lung Cancer, Prostate Cancer-Associated Transcript 1, MicroRNA-129, ATP-Binding Cassette Sub-Family B member 1

Send correspondence to: Huanwen Chen, Cardiothoracic surgery, the First Affiliated Hospital of Chongqing Medical University, No.1 Youyi Road, Yuzhong District, Chongqing 400016, China, Tel: 86-023-89012238, Fax: 02365102346, E-mail: chenhwcqykdx1204@163.com