Li Yadong, Zhang Jinsong, Hong Suling
Department of Otolaryngology, The First Affiliated Hospital of Chongqing Medical University, No.400016, Chongqing, China,
Med Oral Patol Oral Cir Bucal. 2014 Jul 1;19(4):e313-9. doi: 10.4317/medoral.19076.
The purpose of this study is to confirm that ANO1 correlates with occurrence and metastasis of OSCC.
Immunohistochemistry was used to detect the expression of ANO1 in 160 specimens of OSCC and normal tissues. Lentiviral silencing ANO1 was used in SCC-25 cell line to study the cell migration and cell detachment.
Immunohistochemical staining revealed that ANO1 was expressed in a large majority (132 out of 160, 82.5%) of OSCC specimens and that the rate of ANO1 expression in OSCC was significantly higher than that of normal tissue (P<0.05); The rate of ANO1 expression was higher in metastatic tumors than in non-metastatic tumors, and the difference was significant (P<0.05). The results of cell migration assay showed that the percentage of cells through the membrane was 26.61 ±0.81 in assay group, and 54.26 ±3.74 in control group, respectively (t=-16.22,P<0.0001). The results of cell detachment assay showed that the percentage of cells detachment was 37.42 ±0.90 in assay group, and 87.38 ±1.59 in control group, respectively (t=-62.34, P<0.0001). The results of wound healing assay showed the assay group had a reduced migration rate compared with the control group in 32 h (F=1038.78, P<0.0001). Wound closure was no significantly different between the assay and control cells when DIDS was used in wound healing assay (F=4.61,P>0.05).
Our study shows that abnormal expression of ANO1 correlated with the occurrence and metastasis of OSCC in clinical specimens and that silencing ANO1 greatly reduced migration ability of scc-25 cells. Calcium activated chloride channel activity of ANO1 promoted the cell migration. Thus, ANO1 could represent a new diagnostic biomarker and a potentially important therapeutic target of OSCC.
本研究旨在证实ANO1与口腔鳞状细胞癌（OSCC）的发生和转移相关。
采用免疫组织化学法检测160例OSCC标本及正常组织中ANO1的表达。利用慢病毒沉默SCC-25细胞系中的ANO1，以研究细胞迁移和细胞脱离情况。
免疫组织化学染色显示，绝大多数（160例中的132例，82.5%）OSCC标本中表达ANO1，且OSCC中ANO1的表达率显著高于正常组织（P<0.05）；转移瘤中ANO1的表达率高于非转移瘤，差异有统计学意义（P<0.05）。细胞迁移试验结果显示，试验组穿膜细胞百分比分别为26.61±0.81，对照组为54.26±3.74（t=-16.22，P<0.0001）。细胞脱离试验结果显示，试验组细胞脱离百分比分别为37.42±0.90，对照组为87.38±1.59（t=-62.34，P<0.0001）。伤口愈合试验结果显示，试验组在32小时时的迁移率低于对照组（F=1038.78，P<0.0001）。在伤口愈合试验中使用二苯乙烯二磺酸（DIDS）时，试验组和对照组细胞的伤口闭合情况无显著差异（F=4.61，P>0.05）。
我们的研究表明，临床标本中ANO1的异常表达与OSCC的发生和转移相关，沉默ANO1可显著降低SCC-25细胞的迁移能力。ANO1的钙激活氯离子通道活性促进了细胞迁移。因此，ANO1可能是一种新的诊断生物标志物和OSCC潜在的重要治疗靶点。
