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


Affiliations

  • 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

Abstract

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.

Keywords

miRNAs, miR-141, Stem-Loop RT PCR

Subject Discipline

Pharmaceutical and Medical Biotechnology

Full Text:

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