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

Li Y, Zhang J, Hong S - Med Oral Patol Oral Cir Bucal (2014)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Otolaryngology, The First Affiliated Hospital of Chongqing Medical University, No.400016, Chongqing, China, llxxyydd2006@sina.com.

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.

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Expression of ANO1 in clinical specimens and the scc-25 cells. A: ANO1 is negative in oral normol tissure by immunohistochemical staining SP×200; B: ANO1 is positive in oral cancer tissure by immunohistochemical staining SP×200; C: ANO1 is negative in oral cancer tissure by immunohistochemical staining SP×200; D: The RT- PCR analysis of ANO1 mRNA levels in scc-25 cells after infection with lentivirus,the expression levels of ANO1 mRNA were (0.14±0.01) in assay group,(0.32±0.02) in control group, (Student’s t-test, t=-24.55, P<0.0001); E: The Western blots analysis of ANO1 protein expression in scc-25 cells after infection with lentivirus,the protein expression levels of ANO1 were(0.22±0.08) in assay group,(1.16±0.09)in control group , (t=-19.66, P<0.0001).
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Figure 1: Expression of ANO1 in clinical specimens and the scc-25 cells. A: ANO1 is negative in oral normol tissure by immunohistochemical staining SP×200; B: ANO1 is positive in oral cancer tissure by immunohistochemical staining SP×200; C: ANO1 is negative in oral cancer tissure by immunohistochemical staining SP×200; D: The RT- PCR analysis of ANO1 mRNA levels in scc-25 cells after infection with lentivirus,the expression levels of ANO1 mRNA were (0.14±0.01) in assay group,(0.32±0.02) in control group, (Student’s t-test, t=-24.55, P<0.0001); E: The Western blots analysis of ANO1 protein expression in scc-25 cells after infection with lentivirus,the protein expression levels of ANO1 were(0.22±0.08) in assay group,(1.16±0.09)in control group , (t=-19.66, P<0.0001).

Mentions: For immunohistochemical analysis, staining in all specimens was restricted to the tumor cells, with no staining of the stroma. No staining was detected in normal mucosal. Protein expression of ANO1 was detected in 132 (82.5%) of 160 cancer tissues. The rate of ANO1 expression was higher in metastatic tumors than in non-metastatic tumors, and the difference was statistically significant (Chi-square test, P<0.05). The rate of ANO1 expression increased in accordance to clinical stage (P<0.05). No significant differences between the rate of ANO1 expression and differentiation, sex and age were observed (P>0.05) (Fig. 1, Table 1).


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

Li Y, Zhang J, Hong S - Med Oral Patol Oral Cir Bucal (2014)

Expression of ANO1 in clinical specimens and the scc-25 cells. A: ANO1 is negative in oral normol tissure by immunohistochemical staining SP×200; B: ANO1 is positive in oral cancer tissure by immunohistochemical staining SP×200; C: ANO1 is negative in oral cancer tissure by immunohistochemical staining SP×200; D: The RT- PCR analysis of ANO1 mRNA levels in scc-25 cells after infection with lentivirus,the expression levels of ANO1 mRNA were (0.14±0.01) in assay group,(0.32±0.02) in control group, (Student’s t-test, t=-24.55, P<0.0001); E: The Western blots analysis of ANO1 protein expression in scc-25 cells after infection with lentivirus,the protein expression levels of ANO1 were(0.22±0.08) in assay group,(1.16±0.09)in control group , (t=-19.66, P<0.0001).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4119304&req=5

Figure 1: Expression of ANO1 in clinical specimens and the scc-25 cells. A: ANO1 is negative in oral normol tissure by immunohistochemical staining SP×200; B: ANO1 is positive in oral cancer tissure by immunohistochemical staining SP×200; C: ANO1 is negative in oral cancer tissure by immunohistochemical staining SP×200; D: The RT- PCR analysis of ANO1 mRNA levels in scc-25 cells after infection with lentivirus,the expression levels of ANO1 mRNA were (0.14±0.01) in assay group,(0.32±0.02) in control group, (Student’s t-test, t=-24.55, P<0.0001); E: The Western blots analysis of ANO1 protein expression in scc-25 cells after infection with lentivirus,the protein expression levels of ANO1 were(0.22±0.08) in assay group,(1.16±0.09)in control group , (t=-19.66, P<0.0001).
Mentions: For immunohistochemical analysis, staining in all specimens was restricted to the tumor cells, with no staining of the stroma. No staining was detected in normal mucosal. Protein expression of ANO1 was detected in 132 (82.5%) of 160 cancer tissues. The rate of ANO1 expression was higher in metastatic tumors than in non-metastatic tumors, and the difference was statistically significant (Chi-square test, P<0.05). The rate of ANO1 expression increased in accordance to clinical stage (P<0.05). No significant differences between the rate of ANO1 expression and differentiation, sex and age were observed (P>0.05) (Fig. 1, Table 1).

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Otolaryngology, The First Affiliated Hospital of Chongqing Medical University, No.400016, Chongqing, China, llxxyydd2006@sina.com.

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.

Show MeSH
Related in: MedlinePlus