circ_0000326靶向miR-567对口腔鳞状细胞癌HSC3细胞增殖、侵袭及迁移的调控效应

doi:10.19439/j.sjos.2023.06.006

上海口腔医学 ›› 2023, Vol. 32 ›› Issue (6): 590-596.doi: 10.19439/j.sjos.2023.06.006

circ_0000326靶向miR-567对口腔鳞状细胞癌HSC3细胞增殖、侵袭及迁移的调控效应

刘文静¹, 李梦琦², 崔祥³, 汪俊兰⁴

1.合肥市第二人民医院（安徽医科大学附属合肥医院）口腔科,安徽合肥 230011;
2.江苏大学附属医院口腔科,江苏镇江 210031;
3.南京康贝佳口腔医院,江苏南京 210000;
4.南京医科大学附属金陵医院口腔科,江苏南京 210000

收稿日期:2023-06-09 修回日期:2023-08-15 出版日期:2023-12-15 发布日期:2024-01-12
通讯作者: 汪俊兰,E-mail：1443553608@qq.com
作者简介:刘文静（1991-）,女,本科,医师,E-mail：kaylauhome@sina.com

Study on the molecular mechanism of circ_0000326 regulating the proliferation, invasion and migration of oral squamous cell carcinoma HSC3 cells by targeting miR-567

LIU Wen-jing¹, LI Meng-qi², CUI Xiang³, WANG Jun-lan⁴

1. Department of Stomatology, The Second People's Hospital of Hefei,Hefei Hospital Affiliated to Anhui Medical University. Hefei 230011, Anhui Province;
2. Department of Stomatology, Affiliated Hospital of Jiangsu University. Zhenjiang 210031, Jiangsu Province;
3. Kangbeijia Stomatological Hospital.Nanjing 210000, Jiangsu Province;
4. Department of Stomatology, Jinling Hospital, Jinling Clinical College of Nanjing Medical University. Nanjing 210000, Jiangsu Province, China

Received:2023-06-09 Revised:2023-08-15 Online:2023-12-15 Published:2024-01-12

摘要/Abstract

摘要： 目的: 探讨circ_0000326调控口腔鳞状细胞癌HSC3细胞增殖、侵袭及迁移的分子机制。方法: 收集2020年3月—2021年6月安徽医科大学附属合肥医院收治的45例口腔鳞状细胞癌（oral squamous cell carcinoma,OSCC）患者的癌组织及癌旁组织标本,采用qRT-PCR检测circ_0000326、miR-567的表达量,CCK-8法、平板克隆形成实验、划痕实验和Transwell实验分别检测细胞增殖、克隆形成、迁移及侵袭能力,双荧光素酶报告实验检测circ_0000326与miR-567的靶向关系,Western印迹法检测E-cadherin、N-cadherin蛋白表达量。采用SPSS 21.0软件包对数据进行统计学分析。结果: circ_0000326在OSCC组织中的平均表达量为4.01±0.29,在癌旁组织的平均表达量为1.00±0.13;miR-567的表达量分别为0.28±0.03和1.00±0.10,差异具有统计学意义。与si-NC组相比,si-circ_0000326组细胞活力显著降低（P<0.05）,细胞克隆形成数显著减少（P<0.05）。与si-NC组相比,si-circ_0000326组中侵袭细胞数、划痕愈合率显著降低（P<0.05）,E-cadherin蛋白表达量显著升高（P<0.05）,N-cadherin蛋白表达量显著降低（P<0.05）。Circ_0000326靶标miR-567。miR-567在pcDNA组的表达量为1.00±0.00,在pcDNA-circ_0000326组的表达量为0.44±0.04, 在si-NC组的表达量为0.99±0.06,在si-circ_0000326组的表达量为2.92±0.25,差异具有统计学意义。与miR-NC组相比,miR-567组细胞活力显著降低、划痕愈合率及N-cadherin蛋白水平显著降低（P<0.05）,细胞克隆形成数和侵袭细胞数显著减少（P<0.05）,E-cadherin蛋白水平显著升高（P<0.05）。与si-circ_0000326+anti-miR-NC组相比,si-circ_0000326+anti-miR-567组细胞活力、划痕愈合率和N-cadherin蛋白水平显著升高（P<0.05）,细胞克隆形成数和侵袭细胞数显著增多（P<0.05）,E-cadherin蛋白水平显著降低（P<0.05）。结论: 干扰circ_0000326表达可通过促进miR-567表达而减弱OSCC细胞增殖、克隆形成、迁移及侵袭能力。

关键词: 口腔鳞状细胞癌, HSC3细胞系, circ_0000326, miR-567, 细胞增殖, 迁移, 侵袭

Abstract: PURPOSE: To explore the molecular mechanism of circ_0000326 regulating proliferation, invasion and migration of oral squamous cell carcinoma HSC3 cells. METHODS: Cancerous tissue and adjacent tissue specimens of 45 patients with oral squamous cell carcinoma (OSCC) admitted to the Second People's Hospital of Hefei from March 2020 to June 2021 were collected. qRT-PCR was used to detect the expression levels of circ_0000326 and miR-567. CCK-8, plate clone formation test, scratch test and Transwell test were used to detect cell proliferation, clone formation, migration and invasion. Dual luciferase reporter experiment was used to detect the targeting relationship between circ_0000326 and miR-567. Western blot was used to quantify E-cadherin and N-cadherin protein. SPSS 21.0 software package was used for statistical analysis. RESULTS: circ_0000326 expression was 4.01±0.29 in OSCC and 1.00±0.13 in paracancerous tissues, while miR-567 expression was 0.28±0.03 and 1.00±0.10, respectively, with significant differences. Compared with the si-NC group, the cell viability and the number of cell clones in the si-circ_0000326 group were significantly decreased(P<0.05). Compared with the si-NC group, the number of invasive cells and scratch healing rate in the si-circ_0000326 group were significantly decreased (P<0.05), the expression level of E-cadherin protein was significantly increased (P<0.05), and the expression level of N-cadherin protein was significantly decreased(P<0.05). Additionally, circ_0000326 targeted miR-567. miR-567 expression was 1.00±0.00 in pcDNA group, 0.44±0.04 in pcDNA-circ_0000326 group, 0.99±0.06 in si-NC group, and 2.92±0.25 in si-circ_0000326 group with significant differences. Compared with miR-NC group, the cell viability, scratch healing rate, the number of cell clones and the number of invasive cells of miR-567 group were decreased, while E-cadherin protein level was increased(P<0.05). Compared with si-circ_0000326+anti-miR-NC group, the cell viability, scratch healing rate, N-cadherin protein level, the number of cell clones and the number of invasive cells of si-circ_0000326+anti-miR-567 group were increased(P<0.05), while E-cadherin protein level was decreased(P<0.05). CONCLUSIONS: Interference with the expression of circ_0000326 could reduce the ability of OSCC cell proliferation, colony formation, migration and invasion by promoting the expression of miR-567.

Key words: Oral squamous cell carcinoma, HSC3 cell line, circ_0000326, miR-567, Cell proliferation, Migration, Invasion

中图分类号:

R739.8

刘文静, 李梦琦, 崔祥, 汪俊兰. circ_0000326靶向miR-567对口腔鳞状细胞癌HSC3细胞增殖、侵袭及迁移的调控效应[J]. 上海口腔医学, 2023, 32(6): 590-596.

LIU Wen-jing, LI Meng-qi, CUI Xiang, WANG Jun-lan. Study on the molecular mechanism of circ_0000326 regulating the proliferation, invasion and migration of oral squamous cell carcinoma HSC3 cells by targeting miR-567[J]. Shanghai Journal of Stomatology, 2023, 32(6): 590-596.

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circ_0000326靶向miR-567对口腔鳞状细胞癌HSC3细胞增殖、侵袭及迁移的调控效应

Study on the molecular mechanism of circ_0000326 regulating the proliferation, invasion and migration of oral squamous cell carcinoma HSC3 cells by targeting miR-567

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