Wang Congjun, Wang Ye, Fu Zhao, Huang Weijia, Yu Zhu, Wang Jiancheng, Zheng Kaitian, Zhang Siwen, Li Shen, Chen Junqiang
Department of Gastrointestinal Gland Surgery, The First Affiliated Hospital of Guangxi Medical University, Nanning, China.
Clinical Research Lab, Guangxi Key Laboratory of Enhanced Recovery After Surgery for Gastrointestinal Cancer, Nanning, China.
Front Oncol. 2022 Apr 22;12:837581. doi: 10.3389/fonc.2022.837581. eCollection 2022.
MicroRNAs (miRNAs) are small noncoding RNA molecules that regulate genetic expression and are also vital for tumor initiation and development. MiR-29b-3p was found to be involved in regulating various biological processes of tumors, including tumor cell proliferation, metastasis, and apoptosis inhibition; however, the biofunction and molecule-level mechanisms of miR-29b-3p inpapillary thyroid carcinoma (PTC) remain unclear.
The expression of miR-29b-3p in PTC samples was tested qRT-PCR. Cellular proliferation was analyzed by CCK-8 and EdU assays, and cellular migratory and invasive abilities were assessed utilizing wound-healing and Transwell assays. In addition, protein expressions of COL1A1, COL5A1, E-cadherin, N-cadherin, Snail, and Vimentin were identified Western blot (WB) assay. Bioinformatics, qRT-PCR, WB, and dual luciferase reporter assays were completed to identify whether miR-29b-3p targeted COL1A1 and COL5A1. In addition, our team explored the treatment effects of miR-29b-3p on a murine heterograft model.
Our findings revealed that miR-29b-3p proved much more regulated downward in PTC tissue specimens than in adjacent non-cancerous tissues. Meanwhile, decreased expression of miR-29b-3p was strongly related to the TNM stage of PTC patients (p<0.001), while overexpression of miR-29b-3p in PTC cells suppressed cellular migration, invasion, proliferation, and EMT. Conversely, silencing miR-29b-3p yielded the opposite effect. COL1A1 and COL5A1 were affirmed as the target of miR-29b-3p. Additionally, the COL1A1 and COL5A1 were highly expressed in PTC tumor samples than in contrast to neighboring healthy samples. Functional assays revealed that overexpression of COL1A1 or COL5A1 reversed the suppressive role of miR-29b-3p in migration, invasion, and EMT of PTC cells. Finally, miR-29b-3p agomir treatment dramatically inhibited Xenograft tumor growth in the animal model.
These findings document that miR-29b-3p inhibited PTC cells invasion and metastasis by targeting COL1A1 and COL5A1; this study also sparks new ideas for risk assessment and miRNA replacement therapy in PTC.
微小RNA（miRNA）是一类小的非编码RNA分子，可调节基因表达，对肿瘤的发生和发展也至关重要。研究发现，miR-29b-3p参与调节肿瘤的多种生物学过程，包括肿瘤细胞增殖、转移及抑制凋亡；然而，miR-29b-3p在甲状腺乳头状癌（PTC）中的生物学功能及分子水平机制仍不清楚。
采用qRT-PCR检测PTC样本中miR-29b-3p的表达。通过CCK-8和EdU试验分析细胞增殖情况，利用伤口愈合试验和Transwell试验评估细胞迁移和侵袭能力。此外，通过蛋白质免疫印迹（WB）试验鉴定COL1A1、COL5A1、E-钙黏蛋白、N-钙黏蛋白、Snail和波形蛋白的蛋白表达。通过生物信息学、qRT-PCR、WB和双荧光素酶报告基因试验确定miR-29b-3p是否靶向COL1A1和COL5A1。此外，本团队还探索了miR-29b-3p对小鼠异种移植模型的治疗效果。
我们的研究结果显示，与相邻的非癌组织相比，miR-29b-3p在PTC组织标本中的表达下调更为明显。同时，miR-29b-3p表达降低与PTC患者的TNM分期密切相关（p<0.001），而在PTC细胞中过表达miR-29b-3p可抑制细胞迁移、侵袭、增殖及上皮-间质转化（EMT）。相反，沉默miR-29b-3p则产生相反的效果。COL1A1和COL5A1被确认为miR-29b-�p的靶标。此外，与相邻的健康样本相比，COL1A1和COL5A1在PTC肿瘤样本中高表达。功能试验表明，过表达COL1A1或COL5A1可逆转miR-29b-3p对PTC细胞迁移、侵袭及EMT的抑制作用。最后，在动物模型中，miR-29b-3p激动剂治疗显著抑制了异种移植瘤的生长。
这些研究结果表明，miR-29b-3p通过靶向COL1A1和COL5A1抑制PTC细胞的侵袭和转移；本研究也为PTC的风险评估和miRNA替代疗法带来了新的思路。
