Development of Stem-Loop Real-Time PCR Technique for miRNA-141 Expression Analysis in Nasopharyngeal Carcinoma


  • Ho Chi Minh City Open University, Faculty of Biotechnology, Department of Pharmaceutical and Medical Biotechnology, Ho Chi Minh City, 700000, Viet Nam
  • Ho Chi Minh City Open University, Faculty of Biotechnology, Department of Pharmaceutical and Medical Biotechnology, Ho Chi Minh City, Viet Nam


Background: The deregulation of miRNA-141 (miR-141) has been reported to be involved in development and progression of many various types of human cancers. Previously, it has been demonstrated that miR-141 served as the oncogenic role in nasopharyngeal carcinoma in different countries. However, in Vietnam, to our knowledge, there are limit study of the types of evaluation of miR-141 expression. Objective: For the aims to find out the novel diagnosis strategies and promising therapies for many human based on the expression of miRNA, we the stem-loop qRT-PCR method described here is to detect mature miRNA-141 in a fast, specific, accurate, and reliable manner. Methods: StL-miR-141, known as stem-loop RT primer is applied for Reversed transcriptase assay, followed by Real-time PCR assay using StL-F, STL-R primer and SYBRGreen. Results: Experimentally, this method was applied in detection of miR-141 in Vietnamese NPC biopsy samples, as the results, 5 of six NPC were positive to miR-141 expression. Moreover, the electrophoresis analysis was performed to evaluate the Specificity. The assay has shown high specific for miR-141 detection. Conclusion: We successfully designed the stem-loop RT primer, StL-F, STL-R primer, might be used as a rapid, simple method for the detectionwith a wider scope to apply this method onto the Vietnamese population.


miRNAs, miR-141, Stem-Loop RT PCR

Subject Discipline

Pharmaceutical and Medical Biotechnology

Full Text:


GLOBOCAN. Estimated cancer incidence, mortality and prevalence worldwide in 2012. 2012.

Mahdavifar N, Ghoncheh M, Mohammadian-Hafshejani A, Khosravi B, Salehiniya H. Epidemiology and inequality in the incidence and mortality of nasopharynx cancer in Asia. Osong Public Health Res. Perspect. 2016; 7(6):360-72. Doi: 10.1016/j.phrp.2016.11.002. PMid: 28053841, PMCid: PMC5194228.

Epstein JB, Jones CK. Presenting signs and symptoms of nasopharyngeal carcinoma. Oral Surg. Oral. Med. Oral. Pathol. 1993; 75(1):32-36. Doi: 10.1016/0030-4220(93)90402p.

Hao SP, Tsang NM, Chang KP, Ueng SH. Molecular diagnosis of nasopharyngeal carcinoma: Detecting LMP-1 and EBNA by nasopharyngeal swab. Otolaryngol Head Neck Surg. 2004; 131(5):651-54. PMid: 15523443.

Liu Y, Zhao R, Wang H, Luo Y, Wang X, Niu W, et al. miR-141 is involved in BRD7-mediated cell proliferation and tumor formation through suppression of the PTEN/ AKT pathway in nasopharyngeal carcinoma. Cell Death Dis. 2016; 7:e2156. https:// PMid: 27010857, PMCid: PMC4823963.

Lee RC, Feinbaum RL, Ambros V. The C. elegansheterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. Cell. 1993; 75(5):843-54.

Ha M, Kim VN. Regulation of microRNA biogenesis. Nat. Rev. Mol. Cell. Biol. 2014; 15(8):509-24. PMid: 25027649.

Bartel DP. MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 2004; 116(2):281-97.

Calin GA, Croce CM. MicroRNA signatures in human cancers. Nat. Rev. Cancer. 2006; 6(11):857-66. PMid: 17060945.

Faraoni I, Antonetti FR, Cardone J, Bonmassar E. miR-155 gene: A typical multifunctional microRNA. Biochim. Biophys. Acta. Biochim. Biophys. Acta. 2009; 1792(6):497-505. PMid: 19268705.

Macfarlane LA, Murphy PR. Micro RNA: Biogenesis, function and role in cancer. Curr. Genomics. 2010; 11(7):537-61. https:// PMid: 21532838, PMCid: PMC3048316.

Du ZM, Hu LF, Wang HY, Yan LX, Zeng YX, Shao JY, et al. Upregulation of MiR-155 in nasopharyngeal carcinoma is partly driven by LMP1 and LMP2A and down regulates a negative prognostic marker JMJD1A. PLoS One. 2011; 6:e19137. https:// PMid: 21541331, PMCid: PMC3082546.

Lovat F, Valeri N, Croce CM. MicroRNAs in the pathogenesis of cancer. Semin. Oncol. 2011; 38(6):72433. PMid: 22082758.

Amin M, Lam AK. Current perspectives of mi-RNA in oesophagealadenocarcinoma: Rolesinpredictingcarcinogenesis, progression and values in clinical management. Exp. Mol. Pathol. 2015; 98(3):411-18. PMid: 25746664.

Lee KT, Tan JK, Lam AK, Gan SY. MicroRNAs serving as potential biomarkers and therapeutic targets in nasopharyngeal carcinoma: A critical review. Crit. Rev. Oncol. Hematol. 2016; 103:1-9. PMid:27179594.

Lao DT, Truong KP, Le HAT.miRNA-141 as the Biomarker for Human Cancers. JPRHC. 2018; 10:42-49. ajprhc/2018/21486.

Li G, Huang M, Cai Y, Ke Y, Yang Y, Sun X. miR 141 inhibits glioma vasculogenic mimicry by controlling EphA2 expression. Mol. Med. Rep. 2018(2); 18:1395-1404.

Lin L, Liang H, Wang Y, Yin X, Hu Y, Huang J, et al. microRNA-141 inhibits cell proliferation and invasion and promotes apoptosis by targeting hepatocyte nuclear factor-3β in hepatocellular carcinoma cells. BMC cancer. 2014; 14:879. PMid: 25425543, PMCid: PMC4289273.

Croce CM, Calin GA. miRNAs, cancer, and stem cell division. Cell. 2005; 122:6-7. Doi: 10.1016/j.cell.2005.06.036. PMid: 16009126.

Chen C, Ridzon DA, Broomer AJ, Zhou Z, Lee DH, Nguyen JT, et al. Real-time quantification of microRNAs by stemloop RT-PCR. Nucleic Acids Res. 2005; 33(20):e179. PMid: 16314309, PMCid: PMC1292995.


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