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doi: 10.1371/journal.pone.0030049. Epub 2012 Feb 20.

Differential expression profile and genetic variants of microRNAs sequences in breast cancer patients

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Differential expression profile and genetic variants of microRNAs sequences in breast cancer patients

Ali A Alshatwi et al. PLoS One. 2012.

Abstract

The technology available for cancer diagnosis and prognosis is not yet satisfactory at the molecular level, and requires further improvements. Micro RNAs (miRNAs) have been recently reported as useful biomarkers in diseases including cancer. We performed a miRNA expression profiling study using peripheral blood from breast cancer patients to detect and identify characteristic patterns. A total of 100 breast cancer patients and 89 healthy patients were recruited for miRNA genotyping and expression profiling. We found that hs-miR-196a2 in premenopausal patients, and hs-miR-499, hs-miR-146a and hs-miR-196a2 in postmenopausal patients, may discriminate breast cancer patients from healthy individuals. In addition, we found a significant association between two microRNA polymorphisms (hs-miR-196a2 and hs-miR-499) and breast cancer risk. However, no significant association between the hs-miR-146a gene and breast cancer risk was found. In summary, the study demonstrates that peripheral blood miRNAs and their expression and genotypic profiles can be developed as biomarkers for early diagnosis and prognosis of breast cancer.

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Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1
Figure 1. Relative expression of microRNAs in breast cancer cases and control subjects.
Total RNA including small RNA was isolated from blood samples from healthy subjects and breast cancer patients using the Blood RNA Isolation Kit (Qiagen, Germany). cDNA was subjected to Real Time PCR using microRNAs assays (Qiagen, Germany). Data derived from quantitative real-time PCR and presented in ΔCT of relative threshold cycles indicating fold changes over normal subjects. Normalization was performed with the small nuclear RNU1A in blood samples. Results are mean values of triplicate experiments. Bars denote standard error (SEM). P-values of the statistical evaluations of miRNA levels were determined by Mann and Whitney-U test.
Figure 2
Figure 2. Relative expression of microRNAs in pre- and post-menopausal breast cancer patients.
Total RNA including small RNA was isolated from blood samples from healthy subjects and breast cancer patients using the Blood RNA Isolation Kit (Qiagen, Germany). cDNA was subjected to Real Time PCR using microRNAs assays (Qiagen, Germany). Data derived from quantitative real-time PCR and presented in ΔCT of relative threshold cycles indicating fold changes over normal subjects. Normalization was performed with the small nuclear RNU1A in blood samples. Results are mean values of triplicate experiments. Bars denote standard error (SEM). P-values of the statistical evaluations of miRNA levels were determined by Mann and Whitney-U test.
Figure 3
Figure 3. Relative expression of microRNAs with respect to genotype pattern.
miRNAs were determined using SYBR Green MicroRNA Assays adopting Quantitative Real Time PCR. Normalization was performed with the small nuclear RNU1A in blood samples. Total RNA including small RNA was isolated from blood samples from healthy subjects and breast cancer patients using the Blood RNA Isolation Kit (Qiagen, Germany). cDNA was subjected to Real Time PCR using microRNAs assays (Qiagen, Germany). Data derived from quantitative real-time PCR and presented in ΔCT of relative threshold cycles indicating fold changes over normal subjects.Results are mean values of triplicate experiments. Bars denote standard error (SEM). P-values of the statistical evaluations of miRNA levels were determined by Mann and Whitney-U test.

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