ANO1 as a marker of oral squamous cell carcinoma and silencing ANO1 suppresses migration of human scc-25 cells

Abstract

Objectives: The purpose of this study is to confirm that ANO1 correlates with occurrence and metastasis of OSCC. Study Design: 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. Results: 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). Conclusions: 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. Key words:Oral squamous cell carcinoma, chloride channel, metastasis.

Novel ANO1 Inhibitor from Mallotus apelta Extract Exerts Anticancer Activity through Downregulation of ANO1

“…Previous studies have shown that pharmacological blockade of ANO1 inhibits cell proliferation of metastatic prostate cancer cells and metastasis of oral cancer cells [ 10 , 28 ]. As shown in Figure 5 A,B, Ani-D2 induced a significant reduction in ANO1 protein levels in a dose-dependent manner.…”

“…ANO1 amplification and overexpression have been reported in various carcinomas [ 7 , 8 , 9 ]. Recent evidence suggests that ANO1 is a potential therapeutic target for cancers such as prostate, oral, breast, and pancreatic cancer [ 8 , 10 , 12 ]. In this study, we performed cell-based screening to identify a novel ANO1 inhibitor from the methanol extract of M. apelta and found that Ani-D2 is a bona fide inhibitor of ANO1.…”

“…ANO1 is widely expressed in various tissues and regulates many physiological activities such as epithelial cell secretion, smooth muscle contraction, cell growth, and nerve cell transmission [ 3 , 4 , 5 ]. In particular, ANO1 is highly amplified and expressed in various carcinomas including oral squamous cell carcinoma (OSCC), prostate, breast, and esophageal cancers and is involved in the proliferation, metastasis, and invasion of cancer cells [ 6 , 7 , 8 , 9 , 10 ].…”

“…Although the underlying mechanism is unclear, anticancer effects of ANO1 inhibition have been reported in several studies [ 11 , 12 , 13 ]. The cell proliferation, metastasis, and invasion of prostate and oral cancer cells were significantly reduced by the inhibition of ANO1 channel function and reduction of ANO1 protein level [ 6 , 10 ]. In addition, downregulation of ANO1 with the treatment of ANO1 shRNA significantly decreased tumor growth in a prostate cancer xenograft mouse model [ 6 ].…”

Los CaCCs: proteínas multifuncionales, de la fisiología a la enfermedad

“…El cáncer es una de las enfermedades con mayor índice de mortalidad anual en todo el mundo. En este contexto, diversas investigaciones científicas muestran evidencia de que en la "maquinaria" celular de varios tipos de cáncer, tales como el de cuello y cabeza (Ayoub et al, 2010;Dixit et al, 2015), pulmón ( Jia et al, 2015), mama (Wu et al, 2015), gastrointestinal (West et al, 2004), pancreático (Sauter et al, 2014), de esófago (Kashyap et al, 2009), próstata (Liu et al, 2012), oral (Li et al, 2014) y uterino (Sah y McCluggage, 2013), el mensaje de construcción de las proteínas CaCCs está activo continuamente provocando que se produzcan más de lo que se producen en una célula normal (Katoh y Katoh, 2003). A esto se le denomina sobreexpresión del canal o proteína, y esta condición es considerada como un factor de potenciación en etapas muy peligrosas e importantes del cáncer como lo son el origen de tumores (Liu et al, 2012) y la migración celular o metástasis (Ayoub et al, 2010).…”