Cancer Management and Research (Oct 2020)
miR-144-3p Contributes to the Development of Thyroid Tumors Through the PTEN/PI3K/AKT Pathway
Abstract
Hui-Ling Cao,1,* Ming-Qiang Gu,2,* Zhuo Sun,3 Zhong-Jian Chen2 1Department of Head and Neck Surgery, Chengwu Hospital Affiliated to Shandong First Medical University, Heze, Shandong, 274200, People’s Republic of China; 2Department of General Surgery, Chengwu Hospital Affiliated to Shandong First Medical University, Heze, Shandong, 274200, People’s Republic of China; 3Department of Oncology, Chengwu Hospital Affiliated to Shandong First Medical University, Heze, Shandong, 274200, People’s Republic of China*These authors contributed equally to this workCorrespondence: Ming-Qiang GuDepartment of General Surgery, Chengwu Hospital Affiliated to Shandong First Medical University, Heze 274200, Shandong, People’s Republic of ChinaTel +86-13573028527Email [email protected]: To explore the expression and related mechanism of miR-144-3p and PTEN in thyroid cancer (TC).Patients and Methods: From February 2018 to November 2019, 62 patients with TC who received treatment in Chengwu Hospital Affiliated to Shandong First Medical University were collected. TC cells and human normal thyroid HTori-3 cells were purchased. The miR-144-3p-inhibitor, miR-144-3p-mimics, empty vector plasmid (miRNA-NC), si-PTEN and sh-PTEN were transfected into B-CPAP and HTh-7 cells. The expressions of miR-144-3p and PTEN in the specimens were tested by qRT-PCR (qP). WB was used to detect the expression of Bcl-2, APR3, N-cadherin, Slug and Bax proteins in the cells. The cell proliferation was detected by MTT, and the cell invasion was tested by Transwell. The apoptosis was detected by flow cytometry (FC).Results: miR-144-3p was highly expressed and PTEN was weakly expressed in the patients’ tissues. The AUC of miR-144-3p and PTEN was > 0.8. miR-144-3p and PTEN were related to TNM stage, lymph node metastasis and differentiation degree of TC patients. The B-CPAP and HTh-7 with the greatest expression differences were selected for transfection. The expression of miR-144-3p in miR-144-3p-inhibitor group was significantly lower than that in NC group (P< 0.01), and that in miR-144-3p-mimics group was significantly higher than that in NC group (p < 0.01). The expression of PTEN in si-PTEN group was significantly lower than that in NC group (P< 0.01), while that in sh-PTEN group was significantly higher than that in NC group (P< 0.01). Silencing miR-144-3p and overexpressing PTEN could inhibit cell proliferation, invasion and promote apoptosis. WB detection uncovered that silencing the miR-144-3p expression and overexpressing PTEN could inhibit the PI3K, Akt, p-AKT, Bcl-2, APR3 and cyclinD1 proteins and promote the up-regulation of Bax expression. Rescue experiments revealed that the cell proliferation, invasion and apoptosis were not different from NC after co-transfection of miR-144-3p-mimics+sh-PTEN and miR-144-3p-inhibitor+si-PTEN into B-CPAP and HTh-7.Conclusion: Inhibition of miR-144-3p expression can up-regulate PTEN and affect cell proliferation, invasion and apoptosis, which may be a potential therapeutic target for TC.Keywords: miR-144-3p, PTEN/PI3K/AKT, EMT, thyroid cancer, biological mechanism
