Wang F, Wang Y F, Su W, Yang H J, Yang H Y
Department of Oral and Maxillofacial Surgery, Peking University Shenzhen Hospital, Shenzhen 518036, China.
Zhonghua Kou Qiang Yi Xue Za Zhi. 2019 Aug 9;54(8):561-567. doi: 10.3760/cma.j.issn.1002-0098.2019.08.011.
To investigate the effects of circular RNA hsa_circ_0063772 on the proliferation, migration and invasion of oral squamous cell carcinoma (OSCC) cells. Thirty-three patients with oral squamous cell carcinoma who were admitted to the Department of Oral and Maxillofacial Surgery, Peking University Shenzhen Hospital from February 2017 to December 2018 were enrolled in this study. Real-time quantative polymerase chain reaction was used to detect the expression level of circular RNA hsa_circ_0063772 in OSCC and corresponding adjacent tissues, OSCC cell lines and human keratinocytes. The expression level of hsa_circ_0063772 was overexpressed in SCC15 and CAL27 cells by using lentivirus, and the effects of this gene on proliferation, migration and invasion of OSCC cells were detected by cell counting assay, scratch assay, Transwell assay, Western blotting and nude mice tumor formation assay. The expression of circular RNA hsa_circ_0063772 in OSCC tissues (9.38±0.34) was lower than that in adjacent tissues (11.30±0.31) (5.20, 0.001), and the expression in OSCC cells (SCC15: 0.12±0.01; SCC25: 0.18±0.02; SCC9: 0.21±0.01; CAL27: 0.13±0.01) was significantly lower than that in human keratinocytes (1.02±0.02) ((SCC15)41.91, (SCC25)29.21, (SCC9)35.16, (CAL27)41.86, 0.001). Overexpression of hsa_circ_0063772 in SCC15 and CAL27 cells can affect tumor cell proliferation, cell counting assay showed that tumor cell proliferation ability in high expression group (SCC15: 0.76±0.01; CAL27: 0.74±0.02) were lower than empty group (SCC15∶1.22±0.04; CAL27: 0.99±0.03; (SCC15)12.58, (CAL27)6.97; 0.05). Transwell migration experiment showed the number of migrated cell in high expression group (SCC15∶148.00±14.57; CAL27: 243.00±13.00) were less than empty group (SCC15: 580.30±42.91; CAL27: 424.70±18.66, 0.01); Transwell invasion assay showed the number of invased cell in high expression group (SCC15: 123.70±6.98; CAL27: 326.00±17.01) were less than empty group (SCC15: 517.70±9.96; CAL27: 454.30±8.09, 0.01). The results of tumor formation in nude mice showed that the tumor volume and mass of the overexpressed group [(306.40±16.51) mm(3); (289.40±11.44) mg] were lower than that of the empty group [(582.60±32.51) mm(3), 7.58, 0.05; (599.60±21.27) mg, 7.58, 0.001]. Compared with adjacent tissues, hsa_circ_0063772 is lowly expressed in oral squamous cell carcinoma. High expression of hsa_circ_0063772 can inhibit the proliferation, migration and invasion of OSCC cells.
为研究环状RNA hsa_circ_0063772对口腔鳞状细胞癌（OSCC）细胞增殖、迁移及侵袭的影响。选取2017年2月至2018年12月于北京大学深圳医院口腔颌面外科收治的33例口腔鳞状细胞癌患者纳入本研究。采用实时定量聚合酶链反应检测环状RNA hsa_circ_0063772在OSCC组织及相应癌旁组织、OSCC细胞系和人角质形成细胞中的表达水平。通过慢病毒转染使SCC15和CAL27细胞中hsa_circ_0063772的表达水平上调，采用细胞计数法、划痕实验、Transwell实验、蛋白质免疫印迹法及裸鼠成瘤实验检测该基因对OSCC细胞增殖、迁移及侵袭的影响。环状RNA hsa_circ_0063772在OSCC组织中的表达水平（9.38±0.34）低于癌旁组织（11.30±0.31）（t = 5.20，P = 0.001），在OSCC细胞（SCC15：0.12±0.01；SCC25：0.18±0.02；SCC9：0.21±0.01；CAL27：0.13±0.01）中的表达水平显著低于人角质形成细胞（1.02±0.02）（（SCC15）t = 41.91，（SCC25）t = 29.21，（SCC9）t = 35.16，（CAL27）t = 41.86，P = 0.001）。hsa_circ_0063772在SCC15和CAL27细胞中的过表达可影响肿瘤细胞增殖，细胞计数法显示高表达组（SCC15：0.76±0.01；CAL27：0.74±0.02）肿瘤细胞增殖能力低于空载体组（SCC15∶1.22±0.04；CAL27：0.99±0.03；（SCC15）t = 12.58，（CAL27）t = 6.97，P < 0.05）。Transwell迁移实验显示高表达组（SCC15∶148.00±14.57；CAL27：243.00±13.00）迁移细胞数少于空载体组（SCC15：580.30±42.91；CAL27：424.70±18.66，P = 0.01）；Transwell侵袭实验显示高表达组（SCC15：123.70±6.98；CAL27：326.00±17.01）侵袭细胞数少于空载体组（SCC15：517.70±9.96；CAL27：454.30±8.09，P = 0.01）。裸鼠成瘤实验结果显示，过表达组肿瘤体积和质量[（306.40±16.51）mm³；（289.40±11.44）mg]低于空载体组[（582.60±32.51）mm³，t = 7.58，P = 0.05；（599.60±21.27）mg，t = 7.58，P = 0.001]。与癌旁组织相比，hsa_circ_0063772在口腔鳞状细胞癌中低表达。hsa_circ_0063772的高表达可抑制OSCC细胞的增殖、迁移及侵袭。
