Abstract：
OBJECTIVE: 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.
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.
摘要：
目的：探讨circ_0000326调控增殖的分子机制，口腔鳞状细胞癌HSC3细胞的侵袭和迁移。
方法：收集2020年3月至2021年6月合肥市第二人民医院收治的45例口腔鳞状细胞癌患者的癌组织及癌旁组织标本。qRT-PCR检测circ_0000326和miR-567的表达水平。CCK-8，平板克隆形成试验，划痕试验和Transwell试验用于检测细胞增殖，克隆形成,移民和入侵。采用双荧光素酶报告基因实验检测circ_0000326与miR-567的靶向关系。Western印迹用于定量E-钙黏着蛋白和N-钙黏着蛋白。采用SPSS21.0软件包进行统计分析。
结果：circ_0000326在OSCC中的表达为4.01±0.29，在癌旁组织中为1.00±0.13，而miR-567的表达分别为0.28±0.03和1.00±0.10,具有显著差异。与si-NC组相比，si-circ_0000326组细胞活力和细胞克隆数显著降低(P＜0.05)。与si-NC组相比，si-circ_0000326组的侵袭细胞数量和划痕愈合率均明显降低（P＜0.05），E-cadherin蛋白表达水平显著升高(P＜0.05),N-cadherin蛋白表达水平显著降低(P＜0.05).此外,circ_0000326靶向miR-567。pcDNA组miR-567表达为1.00±0.00,pcDNA-circ_0000326组0.44±0.04，si-NC组0.99±0.06，si-circ_0000326组和2.92±0.25，差异有统计学意义。与miR-NC组相比,细胞活力，划痕愈合率,miR-567组的细胞克隆数和侵袭性细胞数减少,而E-cadherin蛋白水平升高(P＜0.05)。与si-circ_0000326+抗miR-NC组相比，细胞活力，划痕愈合率,N-钙黏着蛋白水平，si-circ_0000326+抗miR-567组细胞克隆数和侵袭性细胞数均增加(P＜0.05),而E-cadherin蛋白水平降低(P＜0.05)。
结论：干扰circ_0000326的表达可降低OSCC细胞的增殖能力，菌落形成,通过促进miR-567的表达来实现迁移和侵袭。
方法：收集2020年3月至2021年6月合肥市第二人民医院收治的45例口腔鳞状细胞癌患者的癌组织及癌旁组织标本。qRT-PCR检测circ_0000326和miR-567的表达水平。CCK-8，平板克隆形成试验，划痕试验和Transwell试验用于检测细胞增殖，克隆形成,移民和入侵。采用双荧光素酶报告基因实验检测circ_0000326与miR-567的靶向关系。Western印迹用于定量E-钙黏着蛋白和N-钙黏着蛋白。采用SPSS21.0软件包进行统计分析。
结果：circ_0000326在OSCC中的表达为4.01±0.29，在癌旁组织中为1.00±0.13，而miR-567的表达分别为0.28±0.03和1.00±0.10,具有显著差异。与si-NC组相比，si-circ_0000326组细胞活力和细胞克隆数显著降低(P＜0.05)。与si-NC组相比，si-circ_0000326组的侵袭细胞数量和划痕愈合率均明显降低（P＜0.05），E-cadherin蛋白表达水平显著升高(P＜0.05),N-cadherin蛋白表达水平显著降低(P＜0.05).此外,circ_0000326靶向miR-567。pcDNA组miR-567表达为1.00±0.00,pcDNA-circ_0000326组0.44±0.04，si-NC组0.99±0.06，si-circ_0000326组和2.92±0.25，差异有统计学意义。与miR-NC组相比,细胞活力，划痕愈合率,miR-567组的细胞克隆数和侵袭性细胞数减少,而E-cadherin蛋白水平升高(P＜0.05)。与si-circ_0000326+抗miR-NC组相比，细胞活力，划痕愈合率,N-钙黏着蛋白水平，si-circ_0000326+抗miR-567组细胞克隆数和侵袭性细胞数均增加(P＜0.05),而E-cadherin蛋白水平降低(P＜0.05)。
结论：干扰circ_0000326的表达可降低OSCC细胞的增殖能力，菌落形成,通过促进miR-567的表达来实现迁移和侵袭。
