Zhou Ping, Xu Tongdao, Hu Hao, Hua Fei
Department of Endocrine, The Second People's Hospital of Lianyungang, Lianyungang 222000, Jiangsu, China.
Department of Endocrine, The Third Affiliated Hospital of Soochow University, Changzhou 213003, Jiangsu, China.
Int J Endocrinol. 2021 Oct 26;2021:5499963. doi: 10.1155/2021/5499963. eCollection 2021.
Thyroid carcinoma (THCA) is the most frequent endocrine malignancy. Papillary thyroid carcinoma (PTC) is the major subtype of THCA, accounting for over 80% of all THCA cases. LncRNA PAX8-AS1, a tumor suppressor associated with various human cancers, has been reported to be relevant to the regulation of all sorts of cellular processes. The purpose of this study was to verify the role of PAX8-AS1 in PTC.
Three human PTC cell lines (K1, TPC-1, and IHH4) and one normal human thyroid cell line, Nthy-ori3-1, were used in our study. The expression of genes was detected by qRT-PCR. The bioinformatic analysis and luciferase reporter assay were used to confirm the binding relationship of PAX8-AS1 to miR-96-5p, and the targeting relationship of miR-96-5p to PKN2 was also predicted. Cell proliferation and apoptosis capacities were assessed by MTT and flow cytometry, respectively. EdU assay was used to detect cell proliferation. Western blot assay was employed to examine protein expression.
The expression of PAX8-AS1 was decreased in PTC tissues and cells. PAX8-AS1 overexpression inhibited the proliferation of PTC cells and promoted cell apoptosis. In addition, PAX8-AS1 bonds with miR-96-5p, whose downregulation elevated the expression of PKN2 in PTC cells. Importantly, according to the rescue experiments, PKN2 silencing partially reversed the inhibitory effects of PAX8-AS1 expression on PTC cell proliferation and apoptosis.
We found that the PAX8-AS1/miR-96-5p/PKN2 axis was closely related to the progression of PTC, which could be a potential target for treating PTC patients.
甲状腺癌（THCA）是最常见的内分泌恶性肿瘤。乳头状甲状腺癌（PTC）是THCA的主要亚型，占所有THCA病例的80%以上。长链非编码RNA PAX8-AS1是一种与多种人类癌症相关的肿瘤抑制因子，据报道与各种细胞过程的调节有关。本研究的目的是验证PAX8-AS1在PTC中的作用。
本研究使用了三个人类PTC细胞系（K1、TPC-1和IHH4）和一个正常人类甲状腺细胞系Nthy-ori3-1。通过qRT-PCR检测基因表达。采用生物信息学分析和荧光素酶报告基因检测来确认PAX8-AS1与miR-96-5p的结合关系，并预测miR-96-5p与PKN2的靶向关系。分别通过MTT和流式细胞术评估细胞增殖和凋亡能力。采用EdU检测法检测细胞增殖。采用蛋白质免疫印迹法检测蛋白质表达。
PAX8-AS1在PTC组织和细胞中的表达降低。PAX8-AS1过表达抑制PTC细胞增殖并促进细胞凋亡。此外，PAX8-AS1与miR-96-5p结合，miR-96-5p的下调提高了PTC细胞中PKN2的表达。重要的是，根据拯救实验，PKN2沉默部分逆转了PAX8-AS1表达对PTC细胞增殖和凋亡的抑制作用。
我们发现PAX8-AS1/miR-96-5p/PKN2轴与PTC的进展密切相关，这可能是治疗PTC患者的潜在靶点。
