Figures & data
Table 1. Clinicopathological characteristics of LSCC patients in this research
Table 2. The primers used in this study
Figure 1. MiR-26a is increased and FOXO1 is suppressed in human LSCC. (a) MiR-26a expression in LSCC and normal adjacent tissues. (b) The survival rate curve in high miR-26a expression or low miR-26a expression. (c) The link between miR-26a and FOXO1 determined by Pearson correlation. N = 5; **P < 0.05 by two-sided Student’s t test, versus the control group
Figure 2. Sevoflurane induces apoptosis, reduces metastasis and affects EMT in LSCC. (a) Cell viability assessment by CCK-8. Hep-2 and Tu177 cells were treated with either control or Sevoflurane (1 mM, 2, mM, 3 mM or 4 mM) and cell proliferation was determined. (b) Western blot assays were carried out to determine E-cadherin, N-cadherin and vimentin expressions following the treatment of tumor cells with 4 mM of Sevoflurane. (c) Flow cytometry technique was used to examine apoptosis following treatment of LSCC cells with 4 mM of Sevoflurane. (d) Western blots assays were used to determine the apoptosis proteins cleaved caspase-3 and cleaved caspase-9 expression. (e) Wound healing assay was carried out to assess the migration of Hep-2 and Tu 177 cells following treatments with 4 mM of Sevoflurane. (f) Transwell technique was used to determine invasion in LSCC cells following Sevoflurane treatments. N = 5; *P < 0.05, **P < 0.01, ***P < 0.001 by two-sided Student’s t test, versus the control group
Figure 3. Sevoflurane mediates its effects on LSCC via miR-26a targeting. (a) RT-qPCR was used for the assessment of MiR-26a expression following treatment of Hep-2 and Tu177 cells with 4 mM of Sevoflurane, or control. (b) RT-qPCR assay assessing the MiR-26a expression following the treatment of LSCC cell lines with miR-26a-inhibitors or its negative controls. (c) Cell viability assessment by CCK-8 following the transfection of Hep-2 and Tu 177 cells with miR-26a inhibitors or their NC. (d) Flow cytometry assay to examine apoptosis following transfection of LSCC cell lines with miR-26a-inhibitors or their negative controls. (e) Western blots for assessment of cleaved caspase-3 and cleaved caspase-9 apoptotic proteins expressions following transfection of cells with miR-26a inhibitors or their controls. (f) Wound healing assay to study the migration of tumor cells following transfections with miR-26a inhibitors or their controls. (g) Transwell assays was used for determining invasion in LSCC cell lines following the transfection with miR-26a inhibitors. (h) Western blots assays for the assessment of E-cadherin, N-cadherin and vimentin expressions in cells transfected with miR-26a-inhibitors or their controls. N = 5; *P < 0.05, **P < 0.01, ***P < 0.001 by two-sided Student’s t test, versus the control group
Figure 4. MiR-26a binds FOXO1 directly. (a)The complementary sequence between miR-26a and FOXO1. (b) Luciferase activity in wild type luciferase constructs. (c) Luciferase activity in mutant-type luciferase constructs. (d) FOXO1 expression following miR-26a inhibitor transfection. N = 5; ***P < 0.001 by two-sided Student’s t test, versus the control group
Figure 5. Sevoflurane inhibits EMT of LSCC and promotes its apoptosis in vivo. (a) Western blots to determine the expressions of cleaved caspase-3 and cleaved caspase-9 following transfection of LSCC cells with sevoflurane, miR-26a-antagomir or their respective negative controls. (b) Immunohistochemistry staining for N-cadherin and E-cadherin. N = 5; *P < 0.05 by two-sided Student’s t test, versus the control group
