Figure 6.

miR-195 Silencing Reverses the Functions of LINC00355 in HNSCC

HNSCC CSCs were treated with si-LINC00355 in the presence of miR-195 inhibitor or inhibitor-NC. (A) Cell viability measured by the MTT assay. (B) Cell apoptosis detected by flow cytometry. (C) Quantitative analysis for cell apoptosis rate detected by flow cytometry. (D) The expression of LINC00355, miR-195, and the mRNA expression of HOXA10, Vimentin, caspase-3, caspase-9, and E-cadherin determined by qRT-PCR. (E) The protein expression of HOXA10, Vimentin, caspase-3, caspase-9, and E-cadherin determined by western blot analysis. (F) The statistical analysis of the protein expression of HOXA10, Vimentin, caspase-3, caspase-9, E-cadherin, and GAPDH determined by western blot analysis. (G) Cell invasion detected by the Transwell assay (100×). (H) The number of invasive cells. Nude mice were injected with suspension of CSCs treated with si-LINC00355 in the presence of miR-195 inhibitor or inhibitor-NC (n = 12). (I) Tumor weight of mice. (J) Tumor volume of mice at the different time points after injection. (K) Images of tumors at the 35^th day. *p < 0.05 versus the si-LINC00355 + inhibitor-NC group; the measurement data were expressed as mean ± SE. The experiment was repeated three times. Data comparison between two groups was conducted using unpaired t test (B, D, F, H, and I). Cell viability at different time points was analyzed by two-way ANOVA (A), while tumor volume at different time points was analyzed by the repeated-measures ANOVA (J). The T shape line indicates SD.