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. 2017 Nov;38(5):2735-2740.
doi: 10.3892/or.2017.5994. Epub 2017 Sep 25.

miR-146a and miR-146b in the diagnosis and prognosis of papillary thyroid carcinoma

Zhenwei Qiu  1 , Hai Li  1 , Jianwei Wang  1 , Chongbing Sun  2
Affiliations

miR-146a and miR-146b in the diagnosis and prognosis of papillary thyroid carcinoma

Zhenwei Qiu et al. Oncol Rep. 2017 Nov.

Abstract

The present study investigated the relationship between the expression of miR-146a and miR‑146b with the occurrence and prognosis of papillary thyroid carcinoma. Experiments in vitro were also used to explore the effect of the knocked down expression of the miRNAs on growth and migration of papillary thyroid carcinoma cells. A total of 73 patients with papillary thyroid carcinoma admitted to Yidu Central Hospital of Weifang from September 2013 to September 2015 were enrolled in the study. Carcinoma samples were obtained from each patient, and adjacent tissues were used as control samples to determine expression levels of miR-146a and miR146b by semi-quantitative RT-PCR. An analysis was conducted to find possible correlations between the miRNAs expression levels and clinicopathological features in the patients followed up for one year after diagnosis. Additionally, to examine the function of miR-146a and miR‑146b on TPC-1 cells, the expression of miRNAs was knocked down using specific siRNAs. MTT and Transwell assays were used to evaluate cell proliferation and migration, respectively, in the miRNA cell lines. Finally, western blot analysis was used to analyze the expression of IRAK1 in PTC cancer cells. Our results showed that the expression levels of miR-146a and miR-146b in carcinoma tissues were significantly higher than the levels in cancer-free tissues (p<0.01). The relative expression levels of miR-146a and miR-146b in cancerous tissues could be associated with the pathological type and presence or absence of lymph node metastasis (p<0.05). Compared with the siRNA-control cell, MTT and Transwell assays showed that the cell growth and migration of TPC-1 cells were decreased in miR-146a and miR-146b low expression cells (p<0.01). Western blot analysis showed that the expression of IRAK1 in papillary thyroid carcinoma was higher than in adjacent tissue (p<0.01). Based on our findings, the expression of miR-146a and miR-146b correlates with the occurrence and prognosis of papillary thyroid carcinoma, and the expression levels of miR-146a and miR-146b seem to affect the cell proliferation and migration and regulate the expression of IRAK1 protein in cancer cells. Further studies are needed to validate our results to provide new targets for prevention and treatment of papillary thyroid carcinoma.

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Figures

Figure 1.
Figure 1.
Relative expressions of miR-146a and miR-146b in cancer and adjacent tissues of papillary thyroid cancer patients by semi-quantitative RT-PCR. (A) Agarose gel electrophoresis; (B and C) relative expression levels of miR-146a and miR-146b, the relative expression of miR-146a and miR-146b in cancer tissues was significantly higher than in the adjacent tissues in papillary thyroid carcinoma patients (**P<0.01).
Figure 2.
Figure 2.
Relationship between the expression of miR-146a and the survival time. The survival time of the high miR-146a expression group was significantly shorter than that of the normal or low miR-146a expression groups, (P<0.01).
Figure 3.
Figure 3.
Relationship between the expression of miR-146b and the survival time. The survival time of high miR-146b expression group was significantly shorter than that of the normal or low miR-146b expression groups (P<0.01).
Figure 4.
Figure 4.
Semi-quantitative RT-PCR to verify expression levels of miR-146a and miR-146b in TPC-1 cell lines. (A) miR-146a expression; (B) miR-146b expression.
Figure 5.
Figure 5.
Cell proliferation detected by MTT. (A) Effect of low expression of miR-146a on cell proliferation, the number of cells in the miR146a-siRNA group was significantly lower than that in the siRNA-control group (**P<0.01). (B) Effect of low expression of miR-146b on cell proliferation, the number of cells in the miR146b-siRNA group was significantly lower than that in the siRNA-control group (**P<0.01).
Figure 6.
Figure 6.
Cell migration ability detected by Transwell assay. (A) Effect of miR-146b knock-down on the migration of TPC-1 cells; (B) effect of miR-146b knock-down on the migration of TPC-1 cells (*P<0.05, **P<0.01).
Figure 7.
Figure 7.
Western blot detection of IRAKI protein expression levels. (A) Western blot analysis, lanes 1 and 2 show papillary thyroid carcinoma samples; lanes 3 and 4 show the coresponding adjacent tissues. (B) Relative expressions of IRAK1 protein, compared with the adjacent tissues, the expression level of IRAKI protein in the cancer tissues of papillary thyroid carcinoma was significantly decreased (**P<0.01).
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