Overexpression of CD300A inhibits progression of NSCLC through downregulating Wnt/β-catenin pathway

Figures & data

Figure 1 CD300A was downregulated in NSCLC and correlated with prognosis of patients.

Notes: (A) The expression levels of CD300A mRNA in LUAD tissues (n=483), LUSC tissues (n=486) and their corresponding control samples (n=347, n=338) from GEPIA analysis. Red box is NSCLC tissues (T), the black box is normal tissues (N). (B) The survival curves of NSCLC patients from GEPIA analysis. *P<0.05.
Abbreviations: GEPIA, Gene Expression Profiling Interactive Analysis; LUAD, lung adenocarcinoma; LUSC, lung squamous cell carcinoma; NSCLC, non-small-cell lung cancer.

Figure 2 Overexpression of CD300A decreases the viability and proliferation of NSCLC cells.

Notes: (A, B) The expression level of CD300A mRNA in overexpressed and knocked down groups by quantitative reverse-transcription PCR for A549 (A) and H1650 (B) cells, and blank vector or scramble siRNA was used as NC. (C, D) Cell proliferation rate was examined at 0, 24, 48, and 72 h by using CCK8 assay for A549 (C) and H1650 (D) cells with CD300A overexpression or knockdown. (E, F) Colony formation assay was performed for A549 cell (E) and H1650 cell (F) with CD300A OV or KD. CON: cells without any treatment; NC-OV: cells transfected with pcDNA3.1; OV: cells transfected with pcDNA3.1-CD300A; NC-KD: cells transfected with scramble siRNA; KD: cells transfected with siRNA-CD300A. Scare bar: 10 mm. Data were expressed as the mean ± SD from triple independent experiments. *P<0.05, **P<0.01 compared with the NC group.
Abbreviations: CCK-8, Cell Counting Kit-8; h, hours; KD, knockdown; NC, negative control; NSCLC, non-small-cell lung cancer; OV, overexpression.

Figure 3 Overexpression of CD300A decreases migration of NSCLC cells.

Notes: (A, B) After transection for 24 hours, Transwell migration assay was used to assess cell migration of A549 (A) and H1650 (B) cells. NC-OV: cells transfected with pcDNA3.1; OV: cells transfected with pcDNA3.1-CD300A; NC-KD: cells transfected with scramble siRNA; KD: cells transfected with siRNA-CD300A. Scare bar: 100 µm. Data were expressed as the mean ± SD from triple independent experiments. *P<0.05 compared with the NC group.
Abbreviations: KD, knockdown; NC, negative control; NSCLC, non-small-cell lung cancer; OV, overexpression.

Figure 4 Overexpression of CD300A promotes cell apoptosis of NSCLC cells via the mitochondrial apoptosis pathway.

Notes: (A, B) After transfection for 12 hours, A549 cells (A) and H1650 cells (B) were stained with Annexin V-FITC-PI. Flow cytometric analysis was performed to detect cell apoptosis. (C, D) After transfection for 24 hours, Western blot assay was used to examine expression level of apoptosis-related proteins in A549 cell (C) and H1650 cell (D). Data were expressed as the mean ± SD from triple independent experiments. *P<0.05, **P<0.01 compared with the NC group. NC-OV: cells transfected with pcDNA3.1; OV: cells transfected with pcDNA3.1-CD300A; NC-KD: cells transfected with scramble siRNA; KD: cells transfected with siRNA-CD300A.
Abbreviations: FITC, fluorescein isothiocyanate; KD, knockdown; NC, negative control; NSCLC, non-small-cell lung cancer; OV, overexpression; PI, propidium iodide.

Figure 5 Overexpression of CD300A inhibits the Wnt/β-catenin pathway in NSCLC cells.

Notes: (A, B) After transfection for 24 hours, the expression levels of key components of Wnt/β-catenin pathway were examined using Western blot assay in CD300A overexpressed and knocked down A549 (A) and H1650 (B) cells. Data were expressed as the mean ± SD from triple independent experiments. *P<0.05 compared with the NC group. NC-OV: cells transfected with pcDNA3.1; OV: cells transfected with pcDNA3.1-CD300A; NC-KD: cells transfected with scramble siRNA; KD: cells transfected with siRNA-CD300A.
Abbreviations: GAPDH, glyceraldehyde 3-phosphate dehydrogenase; KD, knockdown; NC, negative control; NSCLC, non-small-cell lung cancer; OV, overexpression.