Asian Journal of Pharmaceutical Research and Health Care

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


Affiliations
1 Faculty of Biotechnology, Department of Pharmaceutical and Medical Biotechnology, Ho Chi Minh City Open University, Ward Co Giang, District 1, HCMC, 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 SYBR-Green. 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.
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  • Development of Stem-Loop Real-Time PCR Technique for miRNA-141 Expression Analysis in Nasopharyngeal Carcinoma

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Authors

Thuan Duc Lao
Faculty of Biotechnology, Department of Pharmaceutical and Medical Biotechnology, Ho Chi Minh City Open University, Ward Co Giang, District 1, HCMC, Ho Chi Minh City, Viet Nam
Thuy Ai Huyen Le
Faculty of Biotechnology, Department of Pharmaceutical and Medical Biotechnology, Ho Chi Minh City Open University, Ward Co Giang, District 1, HCMC, 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 SYBR-Green. 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.

References





DOI: https://doi.org/10.18311/ajprhc%2F2019%2F24990