MIR-150 electrochemical detection connected with specific isolation based on magnetic particles

The microRNA (miRNA) belongs to short noncoding RNA molecules (generally 20-25 nt in length) which are able of specific gene regulation expression ¹. These regulatory molecules have significant influence on different biological processes like cell proliferation, development, differentiation, apoptosis and metabolism ². The aberrant expression of various miRNAs is involved in the cell pathological status formation. The abnormal miRNA expression levels were found at number diseases like inflammatory and autoimmune diseases, diabetes, cardiovascular and neurodegenerative diseases and cancer ^3-6. Specific miRNAs expression profiles were identified at various tumors so miRNA are interesting diagnostic biomarkers. Moreover their detection might be helpful to find more about cancer staging, prognosis and/or response to treatment ⁷. Different studies have shown that miR-150 is upregulated in lung cancer tissue and lupus nephritis, downregulated at patients with heart failure, acute myeloblastic leukemia, colorectal cancer and systemic sclerosis ^8-14. Because of miRNAs importance the simple and fast detection method is needed. Therefore, the sensitive electrochemical analysis was combined with specific magnetic particles-based isolation.

Fig. 1 Electrochemical detection parameters optimization. (A) Optimization of accumulation time. (B) Optimization of acetate buffer pH. Selected parameters were highlighted. Results are expressed as average ± standard deviation (number of measurement n = 3).

Fig. 2 SWV peak height dependence on concentration of (A) miR-150 and (B) antisense miR-150. SWV parameters were as follows: time of accumulation 120 s, start potential 0 V, end potential -1.8 V, potential step 5 mV, frequency 280 Hz, amplitude 25.05 mV. Results are expressed as average ± standard deviation (number of measurement n = 3).

Fig. 3 (A) Dependence of the peak height on applied miR-150 concentration using the MPs-based isolation. Inset: the isolated miR-124 voltammograms (SWV conditions as in Fig. 2). Results are expressed as average ± standard deviation (number of measurement n = 3).

1. Bhayani M. K., Calin G. A., Lai S. Y.: Mutat. Res.-Fundam. Mol. Mech. Mutagen., 731, 14 (2012).
2. Zhang Y., Wang Z. J., Gemeinhart R. A.: J. Control. Release, 172, 962 (2013).
3. Dai R., Ahmed S. A.: Transl. Res., 157, 163 (2011).
4. Takahashi P., Xavier D. J., Evangelista A. F., Manoel-Caetano F. S., Macedo C., Collares C. V. A., Foss-Freitas M. C., Foss M. C., Rassi D. M., Donadi E. A., Passos G. A., Sakamoto-Hojo E. T.: Gene, 539, 213 (2014).
5. Corsten M. F., Dennert R., Jochems S., Kuznetsova T., Devaux Y., Hofstra L., Wagner D. R., Staessen J. A., Heymans S., Schroen B.: Circ.-Cardiovasc. Genet., 3, 499 (2010).
6. Minones-Moyano E., Porta S., Escaramis G., Rabionet R., Iraola S., Kagerbauer B., Espinosa-Parrilla Y., Ferrer I., Estivill X., Marti E.: Hum. Mol. Genet., 20, 3067 (2011).
7. Calin G. A., Croce C. M.: Nat. Rev. Cancer, 6, 857 (2006).
8. Zhang N., Wei X., Xu L. J.: FEBS Lett., 587, 2346 (2013).
9. Wang D. T., Ma Z. L., Li Y. L., Wang Y. Q., Zhao B. T., Wei J. L., Qiu X., Zhao X. T., Jin Y. X.: Oncol. Rep., 30, 492 (2013).
10. Zhou H., Hasni S. A., Perez P., Tandon M., Jang S. I., Zheng C. Y., Kopp J. B., Austin H., Balow J. E., Alevizos I., Illei G. G.: J. Am. Soc. Nephrol., 24, 1073 (2013).
11. Goren Y., Meiri E., Hogan C., Mitchell H., Lebanony D., Salman N., Schliamser J. E., Amir O.: Am. J. Cardiol., 113, 976 (2014).
12. Fayyad-Kazan H., Bitar N., Najar M., Lewalle P., Fayyad-Kazan M., Badran R., Hamade E., Daher A., Hussein N., ElDirani R., Berri F., Vanhamme L., Burny A., Martiat P., Rouas R., Badran B.: J. Transl. Med. , 11, (2013).
13. Ma Y. L., Zhang P., Wang F., Zhang H. Z., Yang J. J., Peng J. Y., Liu W. J., Qin H. L.: Gut, 61, 1447 (2012).
14. Honda N., Jinnin M., Kira-Etoh T., Makino K., Kajihara I., Makino T., Fukushima S., Inoue Y., Okamoto Y., Hasegawa M., Fujimoto M., Ihn H.: Am. J. Pathol., 182, 206 (2013).
15. Huska D., Hubalek J., Adam V., Vajtr D., Horna A., Trnkova L., Havel L., Kizek R.: Talanta, 79, 402 (2009).
16. Smerkova K., Hudcova K., Vlahova V., Vaculovicova M., Pekarik V., Masarik M., Adam V., Kizek R.: Int. J. Oncol., in press.

PDF