Research Paper
Silencing of long non-coding RNA CASC15 enhances radiosensitivity of esophageal squamous cell carcinoma through microRNA-140-5p-dependent inhibition of MMP7
Figure 5. LncRNA CASC15 impairs the radiosensitivity of ESCC by decreasing miR-140-5p and elevating MMP7. (A) RT-qPCR to test the transfection efficiency in KYSE270 cells. (B) CCK-8 assay to assess cell viability of KYSE270 cells. (C) Flow cytometry to evaluate cell apoptosis of KYSE270 cells. (D) Western blot analysis to determine the protein level of Fas, Ki-67 and cleaved Caspase 3 in KYSE270 cells. E, Clone formation ability of KYSE270 cells after X-ray radiation. *p < 0.05 vs. KYSE270 cells transfected with 4 Gy + NC mimic; #p < 0.05 vs. KYSE270 cells transfected with 4 Gy + sh-NC; &p < 0.05 vs. KYSE270 cells transfected with 4 Gy + oe-NC + sh-NC; @p < 0.05 vs. KYSE270 cells transfected with 4 Gy + oe-CASC15 + sh-NC. The measurement data were presented as the mean ± standard deviation. Data in panel (A, C, D) were analyzed by one-way analysis of variance, followed by Tukey’s post hoc test. Data in panel B were analyzed by repeated-measures analysis of variance, followed by Tukey’s post hoc test, and data in panel C were analyzed by two-way analysis of variance, followed by Tukey’s post hoc test; the experiment was repeated 3 times. 4 Gy + NC mimic, KYSE270 cells transfected with negative control of mimic and treated with 4 Gy irradiation; 4 Gy + miR-140-5p mimic, KYSE270 cells transfected with miR-140-5p mimic and treated with 4 Gy irradiation; 4 Gy + sh-NC, KYSE270 cells transfected with negative control plasmids of short hairpin RNA and treated with 4 Gy irradiation; 4 Gy + sh-MMP7, KYSE270 cells transfected with short hairpin RNA against matrix metallopeptidase 7 and treated with 4 Gy irradiation; 4 Gy + oe-NC + sh-NC, KYSE270 cells transfected with negative control plasmids of overexpression vector and short hairpin RNA and treated with 4 Gy irradiation; 4 Gy + oe-CASC15 + sh-NC, KYSE270 cells transfected with plasmids overexpressing lncRNA CASC15 and negative control of short hairpin RNA and treated with 4 Gy irradiation; 4 Gy + oe-CASC15 + sh-MMP7, KYSE270 cells transfected with plasmids overexpressing lncRNA CASC15 and short hairpin RNA against matrix metallopeptidase 7 and treated with 4 Gy irradiation.