干扰Hsa_circ_0023642调控miR-653对甲状腺癌细胞生物行为的影响

Regulation effect of miR-653 on biological behavior of thyroid cancer cells by interference with hsa_circ_0023642

目的探讨Hsa_circ_023642对甲状腺癌细胞恶性生物学行为的影响及分子机制。方法选取本院25例甲状腺癌患者的癌组织及癌旁组织,用实时荧光定量PCR(RT-qPCR)检测Hsa_circ_023642和miR-653的表达水平;甲状腺癌细胞SW579分为si-NC组、si-Hsa_irc_023642组、miR-NC组、miR-653组、si-Hsa_irc_023642+anti-miR-NC组、si-Hsa_irc_023642+anti-miR-653组。使用四甲基偶氮唑盐比色法(MTT)检验细胞活性;集落形成数则通过克隆培养试验进行检测;细胞凋亡率使用流式细胞术检验;蛋白的表达情况通过蛋白免疫印迹(Western blot)法测验;通过细胞划痕实验检验划痕愈合率;Transwell检测细胞侵袭数;双荧光素酶报告实验检测Hsa_circ_023642和miR-653的靶向关系。结果甲状腺癌组织中Hsa_circ_023642表达水平升高,miR-653表达水平降低(P <0.05)。干扰Hsa_circ_023642或过表达miR-653,细胞活性降低,集落形成数减少,细胞凋亡率升高,Bcl-2相关X(Bax)指标呈明显的上升趋势,同时B细胞淋巴瘤/白血病-2(Bcl-2)、细胞划痕愈合率以及侵袭细胞数均呈下降趋势(P <0.05)。Hsa_circ_023642靶向调控miR-653;抑制miR-653可逆转干扰Hsa_circ_023642对甲状腺癌细胞增殖、凋亡、迁移侵袭的影响。结论干扰Hsa_circ_023642可通过调控miR-653抑制甲状腺癌细胞的增殖、迁移侵袭,促进细胞凋亡。

Objective To explore the effect of hsa_circ_0023642 on the malignant biological behavior of thyroid cancer cells and its molecular mechanism. Methods The cancer tissues and adjacent tissues of 25 patients with thyroid cancer in our hospital were selected,and the expression level of hsa_circ_0023642 and miR-653 were detected by real-time fluorescent quantitative PCR(RT-qPCR);thyroid cancer cells SW579 were divided into si-NC group,si-Hsa_circ_0023642 group,miR-NC group,miR-653 group,si-Hsa_circ_0023642+anti-miR-NC group,and si-Hsa_circ_0023642+anti-miR-653 group. Tetramethylazolium salt colorimetric method(MTT)was used to detect cell viability;clone formation test to detect the number of colonies formed;flow cytometry to detect cell apoptosis rate;western blot to detect protein expression;cell scratch test to detect scratch healing rate;Transwell to detect the number of cell invasion;and dual luciferase reporter experiment to detect the targeting relationship between Hsa_circ_0023642 and miR-653. Results The expression of Hsa_circ_0023642 in thyroid cancer tissue was increased,and the expression of miR-653 was decreased(P < 0.05). After interference with hsa_circ_0023642 or with over-expression of miR-653,cell viability was decreased;the number of colonies formed was decreased;apoptosis rate was increased;Bcl-2 related X(Bax)expression was increased. Meanwhile,B-cell lymphoma/leukemia-2(Bcl-2)expression,and cell scratch healing rate were decreased,and the number of invasive cells all showed a downward trend(P < 0.05). Hsa_circ_0023642 targeted and regulated miR-653;inhibiting miR-653 can reverse the effect of hsa_circ_0023642 on the proliferation,apoptosis,migration and invasion of thyroid cancer cells. Conclusion Interference with hsa_circ_0023642 can inhibit the proliferation,migration and invasion of thyroid cancer cells by regulating miR-653,and promote cell apoptosis.